Burn injury is a major public health issue, due to the worldwide annual occurrence of 11 million cases that cause more than 300,000 fatalities.^1,2 Research indicates burn injuries should be classified as chronic diseases because their effects on the immune system probably cause long-term morbidity.³ Up to three months of hospitalization, along with permanent deformities and disability, result from the survival of nonfatal burn injuries, according to research.⁴ Wound management that starts soon after injury with debridement and autografting acts as standard medical practice since it minimizes the risk for sepsis along with organ dysfunction. The shortage of available autologous skin when treating extensive burns forces surgeons to select either allogeneic or xenogeneic grafts as interim dressings after performing surgical debridement. The most common origin for these short-term grafts comes from human deceased donors and porcine skin materials.⁵ The application of both dead human grafts and pig material poses risks for the body to reject the grafts and the transmission of infectious diseases. The prohibition against porcine grafts exists in Muslim communities due to cultural as well as religious beliefs. ^5,6

History of Burns

Historical records show burn injuries have tortured human beings across multiple generations of recorded history. Archaeological records show that ancient Egypt documented the use of mud and excrement together with oil and plant extracts, as well as other substances, for treating burns, as shown in the Ebers Papyrus written around 1500 BC. In ancient Greece, Hippocrates championed the use of dressings containing pig fat along with resin and bitumen.⁷ During medieval times, medical interventions relied primarily on experience-based methods. Upon its arrival, the Renaissance brought back abandoned scientific investigation methods. In 1517–1588, Italian surgeon Leonardo Fioravanti achieved nose reattachment through the use of “balsama artificiato,” a pharmaceutical solution. The anecdote demonstrates medieval restoration attempts of burned tissues, which preceded contemporary grafting methods. ⁸  The Rialto fire of 1921 and the Coconut Grove nightclub fire of 1942 resulted in crucial progress for burn management, which became instrumental in defining contemporary perspectives on fire burn pathophysiology.⁹ Burn care has gained remarkable speed during the past five decades through advancements like antimicrobial wound coverings and fluid treatment protocols, and early surgical procedures with artificial skin substitute development.¹⁰ Despite these advancements, burn injuries continue to be a serious global concern, particularly in low-resource settings, where access to modern treatments is often limited.

Types of Burn Injuries and their Impact on Wound Healing

Types of Burns Based on Depth

Burns are classified based on how deeply they penetrate the skin layers, since it affects both the duration of healing and the necessary treatment approach.

Superficial (First-Degree Burns)

These burns affect only the epidermis, resulting in red, dry skin surfaces, causing pain and resembling sunburn symptoms. After cooling and hydration, the burns require healing time between 3-7 days.

Superficial Partial-Thickness (Second-Degree Burns)

These burns involve the epidermis and part of the dermis, which leads to red blistered skin with pain. The healing process takes between 10-14 days, during which time moist bandages may be needed.

Deep Partial-Thickness (Second-Degree Burns)

These burns extend deeper into the dermis, causing pale and moist tissue, which becomes painless because of nerve damage. Such burns heal between 3 and 6 weeks, yet patients very likely face both infection and scarring risks.

Full-Thickness (Third-Degree Burns)

These burns destroy the epidermis and entire dermis, resulting in white or charred, or leathery tissue with no remaining nerve function. Skin grafting of the area is a must, and there is a high risk of complications.

Full Thickness and Tissues Beneath (Fourth-Degree Burns)

The most severe type, these burns penetrate to muscles, bones, and organs, thereby creating black, dead tissue that may need potential amputation of the burned area. Patients normally require surgical measures combined with reconstructive surgery during their treatment. ¹¹

Types of Burns Based on Cause

The nature of burn injuries depends on their origin and magnitude because this determines both therapeutic strategies and recovery results. The selection of proper interventions, together with prevention of long-term complications, depends on understanding these different burn types.

Thermal Burns

Thermal Burns develop from exposure to hot liquids that cause scalds and direct contact with flames that result in flame burns. Superficial dermal burns occur most frequently in children and elderly people when scalds affect them. The combination of deep dermal and full-thickness burns occurs with flame burns, while these injuries sometimes include damage to the respiratory system. Scalds heal more quickly than flame burns, which often need surgical treatment. ^{12 45}

Electrical Burns

The contact with either low-voltage or high-voltage electrical sources leads to electrical burns. The wounds from electrical burns consist of entry and exit points that produce tissue damage levels which depend on the voltage strength and resistance, and the current path. The internal damage from high-voltage injuries requires ongoing observation because these wounds might develop delayed necrosis. ¹³

Chemical Burns

Exposure to acids, alkalis, or industrial chemicals causes chemical burns that frequently occur in workplace accidents, together with household exposures. The depth of penetration from alkali burns exceeds that of acid burns, while hydrofluoric acid stands out as a dangerous substance that causes progressive tissue destruction. Medical treatment for chemical burns includes neutralization with substances like calcium gluconate for hydrofluoric acid burns, while chemical reactions during healing delay the recovery process.¹⁴

Physiological response and complications of burn injuries

Burn injuries initiate multiple physiological processes that affect both the healing of affected tissue wounds and body-wide organ functioning. The responses triggered by burn injuries affect both how deeply tissues become damaged and the likelihood of complications and the duration of recovery. Such uncontrolled injuries result in dangerous local and systemic effects, including multiple organ failure.

Local Response to Burn Injuries

There are three distinct zones in burn injuries, each representing different degrees of tissue damage and healing potential. ¹⁵

Zone of Coagulation

This is the area of maximum damage where protein coagulation leads to complete tissue destruction.

Zone of Stasis

Surrounding the coagulation zone, this area has decreased tissue perfusion. The tissue is at risk of developing irreversible damage unless medical personnel provide correct resuscitation measures. Hypotension combined with infection or edema creates conditions that deteriorate the tissue damage in this area.

Zone of Hyperaemia

The outermost zone, which shows improved perfusion and recovery potential unless it faces severe sepsis or prolonged hypoperfusion.

Proper management of the zone of stasis prevents both widening and deepening of the wound since this area exists as three-dimensional tissue.

Systemic Response to Burn Injuries

A burn injury that affects 30% or more of the total body surface area (TBSA) triggers systemic inflammation, which results in multi-organ dysfunction.¹⁶

Cardiovascular Changes

The inflammation of capillaries leads to protein and fluid loss from blood vessels, which creates edema. The body responds by narrowing blood vessels in both the peripheral areas and the splanchnic region. Myocardial contractility decreases, possibly due to tumor necrosis factor (TNF) release. The body experiences hypotension and end-organ hypoperfusion when fluid escapes through burn wounds and when systemic blood pressure decreases.¹⁷

Respiratory Changes

The activity of inflammatory mediators creates bronchoconstriction, which raises the possibility of blocked airways. The development of adult respiratory distress syndrome (ARDS) occurs in patients with severe burn injuries. The body’s metabolic processes increase threefold during hypermetabolism, thus causing muscle deterioration alongside decreased immune function. The need for early and aggressive enteral feeding becomes essential for burn patients because it helps to reduce catabolic effects while maintaining gut integrity.¹⁸

Immunological Changes

The immune system develops a nonspecific reduction in its activity, which impacts both cell-mediated and humoral immunity. Opportunistic infections, along with sepsis, become more likely due to this condition.¹⁹

Neurological Response

Intensive burns with severe injuries lead to neurological changes that produce burn-induced encephalopathy and delirium, and altered pain processing because of extended inflammatory signals. The combination of chronic pain syndromes and post-traumatic stress disorder (PTSD) frequently affects patients who experience severe burns.²⁰

Conventional Burn Treatments: Approaches, Limitations, and Future Considerations

Wound treatment for burns requires immediate appropriate care, which both mitigates harm and accelerates recovery. Standard treatments encompass several key interventions:

Cooling of Burned Areas

Burn victims need immediate cold application on affected areas to decrease tissue destruction and minimize discomfort. Ice or extremely cold substances should not be used in burn treatments because they can cause vasoconstriction and secondary tissue damage. Water should be used as a cooling agent at temperatures ranging from 10°C to 20°C, according to the research. ²¹

Fluid Resuscitation

The loss of substantial fluids through burns leads to hypovolemia before causing shock in patients. Medical practitioners use the Parkland formula, introduced in 1968, to determine fluid requirements throughout the 24-hour post-trauma period during which they administer lactated Ringer’s solution for burn shock prevention. ²²

Infection Control

The risk of infection remains high for burn wounds because improper management can lead to sepsis development. Strict infection control measures, including aseptic dressing techniques and careful observation of wound development, are crucial. Doctors should use antibiotics only for confirmed infections or perioperative prophylaxis, since systemic antibiotic prevention is discouraged to stop antibiotic-resistant organisms and avoid secondary infections. ^{23, 24}

Nutritional Support

After severe burn injury, the body enters a state of extreme catabolism, which develops through the combined effects of catecholamines, cortisol, and inflammatory cytokines. After the injury, the body starts this response within 24-48 hours until it reaches its peak at around two years. The stress response after a burn injury produces various negative effects, including muscle tissue reduction and delayed wound closure, and insulin system problems with weakened immunity function.²⁵ Early nutritional support helps patients overcome this phase while accelerating their wound healing process and preventing infections. Wound healing and recovery strongly benefit from proper nutrition. Clinicians need to manage macronutrient intake carefully in order to prevent metabolic problems, including hyperglycemia and immunosuppression. ^23,26

Surgical Intervention (Early Excision and Grafting)

The medical procedure includes immediate removal of dead tissue through early excision combined with grafting, which remains the standard therapy to advance healing while minimizing the dangers of infection. Patients receive autografts as their preferred choice because autografts use their skin, while skin grafting practices face the challenge of donor site restrictions when treating large burns. ²⁶

Limitations of Existing Therapies

While medical progress has been made, medical treatments for burns continue to carry significant restrictions. Standard fluid resuscitation formulas fail to precisely determine individual fluid requirements, thus causing either inadequate or excessive resuscitation, which creates the risk of complications between hypoperfusion and fluid overload. Medical teams encounter enhanced difficulties in infection control because antibiotic-resistant organisms continue to spread; therefore, practitioners need to implement both antibiotics with caution alongside rigorous infection prevention standards to minimize complications. Special care must be taken to maintain proper macronutrient proportions, as excessive carbohydrates can lead to hyperglycemia, increased inflammation, decreased muscle strength, and high-fat consumption can promote lowered immunity and increased susceptibility to infections. Surgical graft procedures dealing with burn patient injuries encounter two major complications that increase the difficulty of healing following surgical reconstruction, and suffer from a scarcity of donor sites. ²⁶

Why Tilapia Skin was Chosen for Burn Care?

Nile tilapia (Oreochromis niloticus) skin has gained recognition as an effective biomaterial dressing for burns due to its clinical applications. ²⁷ It contains a non-infectious microbiota and a human skin-like morphological structure, with a higher composition of Type I and Type III collagen that exceeds that found in human skin.^28,29 It is essential for tissue repair and faster recovery. Its tensile strength, moisture retention, and biocompatibility make it an ideal wound dressing. ²⁹

Tilapia skin also possesses high levels of omega-3 fatty acids, which provide antioxidant and anti-inflammatory benefits, further accelerating the healing process.³⁰ Additionally, marine peptides from tilapia skin collagen closely resemble human collagen in the extracellular matrix, containing eight essential and nine non-essential amino acids that enhance tissue compatibility.³¹

In Brazil, tilapia skin is a cost-effective alternative to human and pig skin, offering an affordable and widely available treatment for burns. ²⁸ Tilapia skin is an ideal skin graft due to its superior collagen content, providing both Type I and Type III collagen at levels that exceed those found in human skin. Its tensile strength is greater than that of human skin, and it retains more moisture, making it an optimal choice for wound healing.³³ Clinical studies have demonstrated that burns treated with tilapia skin heal faster, with reduced pain and fewer dressing changes, improving patient comfort.^27,31 Its high moisture content and collagen composition create an optimal environment for recovery, shortening healing time by several days. Patients receive two benefits from tilapia skin dressings, i.e., decreased pain experience leading to better comfort levels and fewer dressing changes, which leads to faster healing rates^{. 32}

With its structural similarity to human skin, superior collagen content, and therapeutic properties, tilapia skin stands out as a promising biomaterial for burn treatment.

Tilapia Skin as a Xenograft

A xenograft is a procedure of transplanting body tissue or organs between different species. In burn treatment, the application of xenografts provides brief protective coverage to burns that defends the wounds from infections and allows healing to progress. Research has proven that fish skin made from tilapia operates well as a biological dressing. This choice prevails over xenografts derived from pigs as well as cattle, and human cadavers since it addresses several concerns. Xenografts from mammalian resources increase the danger of disease transfer and provoke immune reactions within the body. The use of these dressing materials creates issues with some religious faiths and also costs higher prices than alternatives, while being monitored and not accessible in all locations.³³ The properties of tilapia skin allowed it to tightly adhere to wound beds while decreasing the number of dressing changes and the amount of anaesthesia needed, which benefited patients and healthcare staff through work reduction. The use of tilted fish skin offered economical benefits as an effective burn wound treatment for superficial partial thickness injuries.³⁴

Preparation of Tilapia Skin for Medical Use

Tilapia skin, due to its structural and biological similarity to human skin, has made it a promising alternative option for burn skin grafts. Multiple advantages occur because of tilapia skin use, primarily stopping infections and decreasing fluid loss, and improving healing times. For security and effectiveness purposes, the sterilization procedure must thoroughly remove all dangerous bacteria and contaminants from tilapia skin before its use^{. 35}

Sterilization Methods

To make tilapia skin safe for medical applications, two primary sterilization methods are used

Chemical Sterilization

Chemical sterilization is the first step in preparing tilapia skin. Using antimicrobial agents and preservatives helps eliminate pathogens from tilapia skin without harming tissue structures. The commonly used agents include.

Chlorhexidine Di Gluconate (2%)

Chlorhexidine di gluconate exists as a 2% solution, used to sterilize tilapia skin primarily during the initial stages of the process. As a broad antimicrobial agent, chlorhexidine gluconate exhibits a broad-spectrum antimicrobial ability against both bacteria and fungi. It shows low toxicity, non-corrosive properties, as well as safe performance on biological tissues, ideal for the first stage of sterilization processes ³⁵

Glycerol (50%, 75%, or 99%)

Acts as a preservative to preserve tissue hydration levels by preserving biological tissue structure. The prepared skin benefits from glycerol as it carries antibiotics and antifungal agents, which improve antimicrobial effectiveness. ³⁵

Sterile Saline

Used as a gentle washing agent to remove any residual chemicals, ensuring that the skin is thoroughly cleaned and free from contaminants. It also helps in maintaining the structural integrity of the tissue. ³⁵

Penicillin/Streptomycin/Fungisol (1%) Solution

A treatment mixture of antibiotics and antifungal agents, used for the prevention of bacterial and fungal skin contamination. This keeps the skin pathogen-free, helps reduce potential infections while supporting proper wound recovery. ³⁵

Radio sterilization

After chemical sterilization, tilapia skin is packaged in plastic envelopes until it reaches a cobalt-60 irradiator for exposure to doses of 25 or 30 kGy.   Scientific studies prove that the sterilization method preserves both the collagen structure as well as all skin properties unaltered. Microbiological testing, histological analysis, and tensiometric properties analysis show that Nile Tilapia skin procurement by chemical or radiosterilization methods prepares the skin for biological dressing usage without notable changes. ^34,35

Application of Tilapia Skin in Burn Treatment

Surgical Technique

The surgical application of tilapia skin for medical purposes combines tissue from tilapia fish with wounds or skin lesions to promote new skin development. The process is described in detail below.^36-38

Figure 1: Application process of Tilapia skin in Burn Care.     Click here to view Figure

Aftercare

The following is a description of some of the aftercare that should be followed:

Bandage

A bandage needs to be placed on top of the tilapia skin graft, both for protection and to maintain its proper position. The medical staff must check dressing placement daily to prevent either tight or loose conditions. ³⁹

Pain control

It is common for patients to experience pain during their healing period. The physician provides medication treatments for pain relief. Individuals recovering from surgical procedures can find relief through the usage of cold compresses since they decrease pain as well as swelling. ⁴⁰

Cleaning and wound care

To prevent infection, the graft site should be kept both clean and dry. Minimal infection will occur when patients keep their graft site both clean and dry. The physician will demonstrate step-by-step instructions about cleaning and dressing, replacing methods. The medical staff may use topical ointments or creams as part of the healing process.⁴¹

Avoid strenuous physical activity

The Patient needs to remain inactive with no sports or demanding activities until their physician allows them back to normal physical activity. Graft healing usually needs stability, so any unnecessary movements should be limited since they could slow down recovery time. ³⁹

Nutrition management

The process of healing requires proper nutrition for it to happen quickly while ensuring its optimal results. A person should eat a balanced diet that includes healthy proteins, vitamins, and essential minerals. Alcohol consumption and smoking should be eliminated from a person’s regimen because they create a negative impact on healing processes. ⁴⁰

Follow-up monitoring

Regular evaluations by the physician determine the patient’s healing and recovery progress. For verification of proper healing, clinical tests such as blood tests or imaging will be conducted^{. 41}

Scientific Evidence Supporting Tilapia Skin In Burn Treatment

Research studies about Tilapia skin dressing effectiveness for burn wounds show promising outcomes that match or exceed traditional therapeutic approaches.

Comparable Healing to Standard Treatments

The Phase II randomized controlled trial (RCT) performed by Lima Júnior et al.³⁴ during 2019 investigated Nile Tilapia skin as an occlusive xenograft for burn wound care. The research evaluated Tilapia skin against silver sulfadiazine cream as a conventional therapy for treating small burns.

The research included 62 participants while monitoring vital clinical wound parameters, including healing speed, together with pain levels and dressing requirements, and medicine consumption. Patients receiving Tilapia skin treatment healed their wounds at a faster rate compared to subjects under regular clinical protocols. Specifically, complete repithelialization occurred in:

Outpatients: 9.77 days (Tilapia skin) vs. 11.20 days (silver sulfadiazine).

Inpatients with superficial burns: 10.56 days (Tilapia skin) vs. 11.70 days (silver sulfadiazine).

Inpatients with deep partial-thickness burns: 18.10 days (Tilapia skin) vs. 21.30 days (silver sulfadiazine).

Applying Tilapia skin to burn injuries led to less necessity of dressing maintenance, which improved both patient comfort and healthcare resource availability. Patient ratings on the Visual Analogue Scale (VAS) showed lower pain intensity among subjects using Tilapia skin for burn treatment, particularly among patients with severe burns during hospital stay^.34

Rapid Re-Epithelialization and Safety in Partial-Thickness Burns

In 2019, Lima Júnior et al.⁴² documented the implementation of Tilapia skin as a xenograft on 23-year-old gunshot survivors who experienced partial-thickness burns. The subject presented with proper upper limb superficial burns and deep partial-thickness burns on the left upper limb. Tilapia skin treatment did not need additional dressing changes due to its effective sticking properties with the wound surface. A 12-day period was necessary for superficial burn healing, alongside deep burn healing, which took 17 days. No adverse effects emerged, and the biomaterial proved its excellent compatibility with human tissue. This research demonstrates that Tilapia skin functions as an innovative burn dressing, offering both superior availability and ease of application and exercise. 

Look and feel comfortable together with reduced hospital staff workload, supporting the cost-effective nature of this approach as an alternative to traditional methods of treatment. ⁴²

Accelerated Wound Healing and Pain Reduction

The study performed by Lima Júnior et al.³⁴ utilized a Phase II randomized controlled trial to evaluate Nile Tilapia skin xenografts for burn treatment against silver sulfadiazine cream (SSDC) as the control. A total of 62 participants took part in the research to evaluate critical clinical data points, including wound healing duration and pain ratings, as well as dressing change requirements and medicine use.

The study showed that Tilapia skin decreased healing time for every participant in the research groups:

Outpatients with superficial burns: Healing in 9.77 days (Tilapia skin) vs. 11.20 days (SSDC).

Inpatients with superficial burns: Healing in 10.56 days (Tilapia skin) vs. 11.70 days (SSDC).

Inpatients with deep partial-thickness burns: Healing in 18.10 days (Tilapia skin) vs. 21.30 days (SSDC).

The application of Tilapia skin provided additional pain relief benefits. Patients treated with Tilapia skin reported significantly lower pain scores on the Visual Analogue Scale (VAS) compared to the SSDC group, especially from the second to eighth evaluation visits for deep burn patients. Patients needed less analgesic medication, and they used considerably lower amounts of ketamine and fentanyl during the treatment period. The requirement for dressing changes decreased significantly because of the Tilapia skin application. The use of Tilapia skin required patients to experience 60-70% less dressing requirement than SSDC patients, minimizing patient discomfort and healthcare workload.

This research has established that Tilapia skin serves dual purposes by enhancing healing time while reducing pain and burn-related discomfort, so that it proves to be both a viable and cost-effective alternative to conventional burn treatments^{. 34}

Superior Moisture Retention and Comfort

A 2022 case series authored by Putri et al. evaluated Tilapia skin xenografts against paraffin gauze for full-thickness burn dressing after burn patients underwent excisional debridement. Research conducted in Dr. Cipto Mangunkusumo Hospital’s Burn Unit of Indonesia examined four patients with 20–40% total body surface area burns who underwent excisional debridement within 96 hours post-burn. Tilapia skin xenografts needed fewer dressing modifications compared to paraffin gauze, as observed in the study results. Durational data showed that patients under Tilapia skin therapy required two fewer dressing changes during the 10-day observation period. The research confirmed both the safety and biocompatibility of Tilapia skin xenografts by showing no allergic reactions and no adverse effects emerged.

Results indicated that patients experienced less pain on the Tilapia-treated side when compared to the side treated with paraffin gauze, which proved that Tilapia provided better patient comfort. Tests showed the paraffin gauze group experienced frequent wound fluid leakage, which did not occur with Tilapia skin since it established a beneficial healing condition.

The research demonstrates that Tilapia skin xenografts present more advantages as burn dressings because they help maintain better moisture balance while needing fewer changes and providing better experiences compared to traditional paraffin-based dressings. ⁴³

The available research shows that tilapia skin represents an appropriate tool to address burn healing when conventional skin grafts cannot be employed. Additional studies are required to completely comprehend the possible advantages of tilapia skin applications while developing official standards for its medical use.

Comparison of Commonly used Wound Dressings for Burn Injuries

This comparison of wound dressings for burn injuries shown in Table 1 is based on data from scientific evidence supporting Tilapia skin in burn treatment, where multiple studies have demonstrated the efficacy of Tilapia skin as a burn dressing. To assess its advantages, Tilapia skin is compared to Silver Sulfadiazine and Paraffin Gauze, two widely used standards in burn care. The analysis investigates clinical performance through investigation, including healing time, infection rate, pain management, scar formation, and patient comfort, alongside ease of use and adverse reactions evaluation to demonstrate superior potential in Tilapia skin usage^{. 48}

Table 1: Comparison of Tilapia Fish Skin to Paraffin Gauze and Silver Sulfadiazine

The data indicate that tilapia fish skin functions as an effective solution for treating burn wounds. It offers faster healing times, reduced infection rates, better pain management, and potentially lower scar formation than when using paraffin gauze and silver sulfadiazine. The benefits of using tilapia fish skin outweigh the handling requirements that the product demands. However, additional investigations are needed to determine both the security and effectiveness of tilapia fish skin use in different patient groups during extended treatment periods.

Clinical Advancements and Future Directions

As research progresses, Tilapia skin is being further optimized for clinical applications through advanced formulations:

Tilapia Fish Skin Collagen Extract Ointment

A 15% Tilapia collagen extract ointment has demonstrated superior wound healing in preclinical studies, significantly reducing burn wound size compared to conventional treatments. The extraction method involves advanced purification techniques, including NaOH and butyl alcohol pre-treatment, acetic acid extraction, and NaCl precipitation, followed by dialysis and freeze-drying steps to extract bioactive collagen, which optimizes its skin regeneration effects. The natural biocompatible ointment differs from traditional burn treatments by providing better hydration and better elasticity, along with decreased scarring effects. The ointment satisfies all pharmaceutical requirements, such as homogeneity along with spreadability and pH stability (pH 4-5), which makes it commercializable worldwide for burn care applications. ⁴⁴

Processed Tilapia Collagen Sponges: Smart Wound Healing with Natural Biomaterials

Collagen sponges derived from Tilapia skin, including Dialyzed Tilapia Skin Collagen Sponge (DTSCS) and Self-Assembled Tilapia Skin Collagen Sponge (STSCS), present a next-generation wound dressing solution, offering improved hemorrhage control and biocompatible healing along with enhanced tissue regeneration properties. The freeze-drying method used to produce these sponges contains no cross-linking chemicals, which results in sponges that exhibit high biocompatibility with less immune response. The sponge structure contains numerous pores, which allow them to sustain favorable wound conditions through effective fluid absorption while speeding up tissue restoration processes. DTSCS and STSCS maintain high levels of porosity (91.25% – 97.38%) that allow them to efficiently absorb water as well as wound exudates, thereby supporting moist healing conditions. Their loose fiber network of collagen serves three wound-healing functions, which include fast cellular movement while promoting tissue regeneration and enhancing nutritional exchange through the network.

The analysis between Tilapia-based sponges showed that STSCS achieved a better outcome compared to conventional bovine collagen sponges, demonstrating faster wound closure, superior biocompatibility, and a reduced inflammatory response by day 14. Scientific investigations demonstrate that STSCS works exceptionally well to stop bleeding in rat models of hepatic and arterial bleeding, within 60 seconds, to stabilize bleeding streams superior to standard dressings and DTSCS. The empirical data suggest Tilapia skin collagen sponges would function as a superior choice to ordinary wound dressing materials by providing three essential features: gradual integration with tissue, accelerated wound restoration, and powerful bleeding arrest capability. These burn care materials are supported by their cost-effectiveness and sustainable production from fish byproducts to create sustainable products. ⁴⁵

Decellularized Tilapia Skin Scaffolds Enhanced with Silver Nanoparticles: A Dual-Action Burn Therapy

The research has established a decellularized fish skin (DFS) scaffold featuring biosynthesized silver nanoparticles (AgNPs) for improving both burn wound healing and protecting against infections. The DFS scaffold maintains its high Type I collagen content to provide tissue regeneration support, and AgNPs offer potent antibacterial properties that stop burn-related infections from developing. The biosynthesized AgNPs obtained from Aloe vera extract underwent characterization to determine size measurements, crystalline properties, and antimicrobial effectiveness levels. The antibacterial potency of nanoparticles at 6-hour incubation reached 29.1 nm, but nanoparticles at 12-hour incubation grew slightly larger at 35.2 nm. The decellularization process yielded a suitable DFS scaffold with intact collagen structure and mechanical properties appropriate for use as a clinical burn dressing. The material exhibits high moisture retention (81.7%) to support wound healing through optimal moist conditions, which helps promote faster re-epithelialization.

Additionally, the scaffold also exhibited high swelling capacity (102.89%), which enabled it to absorb exudates from wounds and safeguard against bacterial proliferation. The AgNP-loaded DFS scaffold proved effective against the major wound pathogens Staphylococcus aureus and Pseudomonas aeruginosa in an antibacterial assay. Tests confirmed that the bactericidal potency of the containing solution reached 50 µg/ml as its minimum inhibitory concentration (MIC). Also, in cytotoxicity studies, the scaffold demonstrated safe characteristics with fibroblast 3T3 cells because it did not cause any toxic effects and simultaneously enhanced cell growth for wound healing purposes. Its antibacterial efficiency, moisture retention, biodegradability, and wound-healing potential make the AgNP-infused DFS scaffold stands as a cost-effective and eco-friendly substitute for synthetic burn dressings. Its biodegradability eliminates the necessity for frequent dressing adjustments, so it is ideal for clinical burn care, especially when healing extends over a protracted period. ⁴⁶

Tilapia Collagen-Based Hydrogel

Collagen, a key biomaterial for wound healing, can be sustainably extracted, and the fish scale origin provides a cost-efficient biocompatible alternative to traditional sources. The researchers used the Taguchi method optimization to discover the optimal parameters for extracting collagen from fish scales with Tris-Glycine buffer. The optimal conditions for maximum collagen yield (17.14 ± 0.05 mg/g) required 0.5 M acetic acid, 100 mL acid volume with 120-minute soaking, followed by the addition of 10 mL Tris-Glycine buffer. The extracted Type I collagen demonstrated high purity as well as structural integrity and included significant levels of glycine (20.98%), proline (15.43%), and hydroxyproline (11.51%), which make it suitable for wound healing and tissue regeneration.

Hydrogels derived from Tilapia constitute a significant advance in burn treatment by delivering both cooling properties and moisturization and bioactive wound dressing benefits. Hydrogels function as instant pain blockers while simultaneously decreasing inflammation and blocking bacterial infections, which are essential components for burn injury treatment. Medical scientists created a durable hydrogel by crosslinking Tilapia collagen with hyaluronic acid, which demonstrates enhanced mechanical strength. These hydrogels demonstrate enhanced properties compared to traditional synthetic hydrogels because they originate from natural sources, while providing improved biocompatibility, reduced allergenicity, and better wound healing results. The eco-friendly collagen extraction technique allows better recovery of collagen materials, reducing waste from the seafood industry. Research findings demonstrate that fish scale collagen shows promise for medical treatments of burns and advances the development of advanced clinical biomaterials. ⁴⁷

Evaluation of Tilapia Skin in Burn Care: Advantages and Drawbacks

Advantages

Accessibility and Cost-Effectiveness

Tilapia fish are inexpensive and widely available, which makes their skin a convenient and affordable option for burn treatment. Tilapia skin, as a commonly available byproduct, helps cut down the costs and problems linked to using different wound dressings. This makes it a particularly cost-effective treatment choice. ²⁷

Bioactive Properties for he:

The skin of tilapia contains hydrated type 3 collagen, which is structurally similar to the human skin. This high collagen content helps dressing to adhere effectively to the wound bed, which results in reduced scarring and minimal side effects.²⁷

Reduced Healing Time and Improved Pain Management

Studies have shown that using tilapia skin on burns provides improved results by reducing healing time. The healing process becomes shorter, because of which means patients experience less pain and require fewer pain medications for treatment. The application of tilapia skin benefits patients because of its natural properties, which reduce discomfort. ²⁷

Dressing-Free Treatment

Unlike traditional bandages and ointments that need to be cleaned and changed frequently, tilapia skin can be left on until the burn heals fully. The patient no longer experiences needless agony and discomfort for the patient.²⁷

Infection Resistance

The antibacterial properties and resistance against infections that occur in Tilapia skin make it an optimal material for treating burn wounds. This advantage sets it apart from other options like pigskin, which lacks similar resistance.²⁷

Preparation-Free and Anesthesia-Free:

Unlike pigskin, which requires both animal fasting and anesthesia during treatment, treatment with tilapia skin does not need animal preparation or anesthesia-free techniques, eliminating extra procedures and associated risks.²⁷

Drawbacks

Odor Sensitivity

Tilapia skin develops a distinct odor as it breaks down over time; this odor might displease both patients and their caregivers. While sterilization reduces the initial microbial load, subsequent breakdown of biological material on the wound can still produce odors that may affect patient comfort and compliance. This is not merely an aesthetic issue; a strong, unpleasant odor can negatively impact a patient’s psychological well-being and comfort during a prolonged recovery period. Professional cleaning methods stand as the main approach to reduce this problem. ²⁷

Sterilization Requirements

Before using fish skin purchased from the market, the skin needs full cleaning and sterilization due to the possibility of muscle tissue attachments. The extra step will extend the duration of treatment procedures. Furthermore, the required sterilization methods, such as glycerolization and gamma irradiation, demand specialized laboratory facilities and protocols. This can present a significant logistical challenge, particularly in low-resource settings where tilapia skin is promoted as most beneficial ²⁷

Adherence Challenges

The adherence of tilapia skin becomes challenging for some burn injury locations, which include areas such as the face, groin, buttocks, neck, genitalia, or axillae. These are often areas of high mobility and complex contours, making it difficult for the relatively stiff, sterilized skin to conform perfectly to the wound bed, potentially leading to graft detachment. This can lead to graft displacement or failure, requiring more frequent monitoring and potentially alternative dressing methods for these specific regions.²⁷    

Conclusion

Burn wound treatment using tilapia fish skin introduces a promising and advantageous solution to traditional skin dressing methods, silver sulfadiazine, and paraffin gauze. Its rich composition of collagen, omega-3 fatty acids, and bioactive compounds contributes to accelerated healing, pain reduction, and potentially minimized scarring, while also providing strong infection resistance. The cost-effectiveness and broad availability of tilapia fish skin prove to be a superior choice for burn care, particularly within healthcare settings where resources are limited. However, challenges related to odor sensitivity, sterilization requirements, and adhesive challenges in particular areas are important considerations for clinical implementation. Dressing changes become fewer, and patient discomfort is reduced through these improvements, which enhance both comfort and treatment effectiveness. While more research is needed to optimize processing, establish universal standardization protocols, and improve application techniques, tilapia skin holds immense potential in revolutionizing burn care, offering an affordable, efficient, and biocompatible alternative that can improve wound healing on a global scale. The application shows promising potential to boost present-day burn care strategies, especially in areas without access to traditional medical tools.

Acknowledgment

All the authors are sincerely acknowledge the valuable guidance and support of  Dr. S.S. Sonawane during the preparation of this review article. Gratitude is also extended to the Department of Quality Assurance, MET Institute of Pharmacy, Nashik, for providing the necessary facilities and academic environment.

Funding Sources

The author(s) received no financial support for the research, authorship, and/or publication of this article.

Conflict of Interest

The authors do not have any conflict of interest.

Data Availability Statement

This statement does not apply to this article.

Ethics Statement

This research did not involve human participants, animal subjects, or any material that requires ethical approval.

Informed Consent Statement

This study did not involve human participants, and therefore, informed consent was not required.

Clinical Trial Registration

This research does not involve any clinical trials.

Permission to reproduce material from other sources

Not Applicable

Author Contributions

Harshada Suren Patil – Wrote the final draft of the work and edited

Kaveri Shantaram Panpatil – Completed all reference work and formatting

Shruti Deepak Shinde – Collected all the data and completed the literature review

Priyanka Sanjay Wabale – Assisted in data collection and performed all data editing

Sandeep Sonawane – Supervised the entire work and provided valuable guidance

References

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45. Wang T, Yang L, Wang G, et al. Biocompatibility, hemostatic properties, and wound healing evaluation of tilapia skin collagen sponges. Journal of Bioactive and Compatible Polymers. 2020;36(1):44-58. doi:1177/0883911520981705
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46. Adhikari SP, Paudel A, Sharma A, et al. Development of Decellularized Fish Skin Scaffold Decorated with Biosynthesized Silver Nanoparticles for Accelerated Burn Wound Healing. Dunne N, ed. International Journal of Biomaterials. 2023;2023:1-18. doi:1155/2023/8541621
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Abbreviations 

ARDS: Adult Respiratory Distress Syndrome;

AgNPs^: silver nanoparticles; DFS, Decellularized Fish Skin;

DTSCS: Dialyzed Tilapia Skin Collagen Sponge;

MIC: Minimum Inhibitory Concentration;

PTSD: Post-Traumatic Stress Disorder;

RCT: Randomized Controlled Trial;

SSDC: Silver Sulfadiazine Cream;

STSCS: Self-Assembled Tilapia Skin Collagen Sponge;

TBSA: Total Body Surface Area;

TNF: Tumor Necrosis Factor;

VAS: Visual Analogue Scale;

BC: Before Christ.

In the modern world, 80% of individuals use herbal medicines to treat common skin conditions. Acne vulgaris is a common inflammatory skin condition that affects 85% of teenagers. It is a developmental stage that primarily affects adolescents between the ages of 18 and 25.¹ Adults aged 35 to 40 years often have lower rates of acne vulgaris, an inflammatory condition of the sebaceous glands.² Pain, redness, and inflammation are some of the symptoms of this illness, and occasionally pus forms. In the modern world, herbal cosmetics are becoming more and more popular. It is also becoming more and more important to treat chronic skin problems with innovative formulations developed in recent years and this condition could be brought on by the skin being exposed to environmental dangers like dust and pollution, eating more fatty meals, the sebaceous glands producing more sebum, bad eating habits, etc. Acne can manifest as inflammatory papules, pustules, nodules, cysts, and other distinctive lesions that can cause pigmentary changes and scarring. Hormonal changes in the body might also result in acne. One prevalent inflammatory skin disorder is acne vulgaris. Acne affects around 90% of teenagers, and half of them still get it as adults. Five percent of women and one percent of men still have lesions at the age of forty. Acne in youngsters is becoming more common, according to an investigation, possibly as a result of pubertal onset.³

A liquid phase is enclosed in a three-dimensional biodegradable gum matrix with a high degree of physical or chemical cross-linking. The intermediate nature of topical gels between solid and liquid materials makes them a great option for a variety of applications. In recent decades topical gels have gained a lot of attention because they are of interest to professionals in industry, research and development, education, drug control administration, and other sectors. In this article, the basic and current developments in topical gels, including their classification and preparation process, are reviewed. A separate section discusses the application of hydrogel in drug delivery systems. There is particular focus on its classification, preparation process, and evaluation criteria.⁴

The goal of the study was to create a polyherbal gel using aloevera and turmeric and add excipients. Members of the Liliaceae family, aloevera is a perennial succulent plant. One name for this plant is “the healing plant.” The use of aloe vera in traditional medicine dates back several centuries, and it has been shown to have immunomodulatory and growth-promoting properties for millennia and one plant with more effectiveness and potency against a variety of skin conditions is aloevera, which also has fewer toxicity and adverse effects. Folklore states that the plant’s gel can be applied straight to the skin to heal.⁵

In contrast to conventional topical preparations and oral administration, gel formulations are utilized to deliver the medicine topically due to their simplicity of application, extended contact time, and reduced side effects. He concluded that the polyherbal gels were made topically using carrageenan-induced rat paw edema and formalin-induced rat paw edema in order to evaluate their anti-inflammatory efficacy. When compared to individual gels, the polyherbal gels demonstrated a synergistic effect that may be helpful in treating local inflammation.⁶

The spice turmeric, or Curcuma longa, which is a member of the Zingiberaceae family, has drawn a lot of interest from the scientific and medical communities as well as from its culinary use. A member of the ginger family (Curcuma longa), turmeric is a rhizomatous herbaceous perennial plant.⁷ Since ancient times, people have been aware of turmeric’s therapeutic benefits. Nevertheless, the capacity to identify the precise mechanism or mechanisms of action as well as the bioactive substances found in the plant.⁸ The primary natural polyphenol present in the rhizome of Curcuma longa, or turmeric, as well as other Curcuma species, is curcumin.⁹ The Curcuma longa’s anti-inflammatory¹⁰, antimutagenic, antibacterial,¹¹ wound-healing, and anticancer qualities have made it a traditional medicinal herb in Asian nations.¹²

Single phase gel is a popular skin care formulation for cosmetic items because of its attractive look. Additionally, organic macromolecules are evenly disseminated throughout a liquid so that there are no discernible borders between the liquid and the dispersed macromolecules. Terminalia chebula Retz. plants have been shown to have antibacterial and antifungal properties, as well as ant inflammatory and wound-healing properties. In light of this, we intended to introduce it as a semisolid external preparation (Extract from Terminalia chebula Retz.) and evaluate its effectiveness for topical anti-inflammatory, wound-healing, antibacterial, and antifungal activity using a gel formulation. Additionally, a review of the literature showed that this formula and stability studies have little scientific backing and therefore, an effort was made to create an aqueous extract gel of Terminalia chebula Retz leaves and conduct stability tests on them.¹³

The methanol leaf extracts of Vitex negundo and Cardiospermum halicacabum may contain luteolin and apigenin, which may be responsible for the topical herbal gel formulation’s anti-arthritic properties. It was demonstrated that the produced formulation F4, which contained 1.5% carbopol 934 and 2% CHME and VNME each, was a promising topical herbal gel for the treatment of arthritis. The use of this formulation can be supported by further clinical study for those with joint inflammatory disorders.¹⁴ Additional clinical research is necessary to assess the healing benefits of the created gel formulation on acne lesions due to the statistically significant in vitro antiacne effects of the formulated gel.¹⁵ 

Skin related problems

Although additional research is required, the herbal gel showed very promising anti-bacterial and anti-fungal action against acne and several skin-related infections in a concentration-dependent way.¹⁶ Almost everyone has acne vulgaris, an inflammatory skin disorder, at some point. The disease’s typical symptoms, which include papules, comedones, pustules, scarring, and nodules, can negatively affect a person’s social and psychological well-being. Allopathic acne treatments are available, but they are costly, have unfavorable side effects, and can lead to antibiotic resistance. The goal of this study is to create and assess topical gels that comprise extracts of Eucalyptus globulus, Allium cepa, and Aloe vera as possible antiacne medications. As a gelling agent, 1% Carbopol 940 was used to create six formulations with the herbal extracts. The plant extracts’ phytochemical makeup was identified. The microbroth dilution method was used to determine the minimum inhibitory concentration (MIC) of the extracts and gels. The formed gels’ homogeneity, color, texture, grittiness, odor, spreadability, extrudability, viscosity, pH, and drug content were among their physicochemical characteristics that were assessed. Triterpenoids, alkaloids, flavonoids, tannins, and coumarins were all present in the plant extracts and in relation to Staphylococcus epidermidis, Staphylococcus aureus, Escherichia coli, Candida albicans, and Pseudomonas aeruginosa, the gel formulations exhibited variable activity at different concentrations. The plant extracts’ phytochemical constituents are most likely what give the gel compositions their antibacterial properties. The 5% Aloe vera-Allium cepa (1:1) combination gel formulation successfully inhibited Staphylococcus epidermidis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Candida albicans, with MICs of 12.50, 25.00, 6.25, 25.00, and 12.50 mg/mL, respectively. In general, the gels had good antibacterial and physicochemical qualities and could be utilized as acne treatments.¹⁷

Skin rashes

Common causes for seeking medical attention include rashes and skin problems. In general, one of the most common disorders in people is skin disease. In the past ten years,^18-20 skin and subcutaneous illnesses have become more common and are now the fourth most common source of nonfatal disease burden globally.²¹ In Skin disorders accounted for 1.79% of the global disease burden in 2013, as determined by disability-adjusted life expectancy for 306 distinct illnesses and accidents. Cellulitis, pyoderma, scabies, and other fungal and viral skin illnesses are among the many skin ailments that are referred to as an important aspect of those conditions.²²

Psoriasis

A chronic inflammatory skin condition that is immune-mediated and affects 2% of white people, psoriasis causes psychological distress, pain, discomfort, and social stigma because of its appearance, which lowers quality of life (QoL).^23,24 Psoriasis is characterized by thicker, inflammatory plaques with silvery scale that are caused by keratinocyte hyperproliferation due to cytokine overactivity.^25-27 Individuals with obvious psoriatic plaques tend to hide their skin and refrain from exercising, which exacerbates osteopenia.²⁸ Many cases of psoriasis are associated with metabolic syndrome.²⁹ The use of biologics to treat severe psoriasis has increased within the last 20 years.^30-32 

Atopic Dermatitis

Atopic dermatitis (AD) is a prevalent, chronic, relapsing, inflammatory skin disease that mostly affects young children. Atopy is characterized as an inherent tendency to create immunoglobulin (Ig) E antibodies in reaction to minute amounts of common environmental proteins such as pollen, house dust mites, and food allergies. The Greek words “derma” (meaning skin) and “itis” (meaning inflammation) are the origins of the word “dermatitis”.³³ The terms dermatitis and eczema are frequently used interchangeably, however eczema is occasionally used to refer to the disease’s acute expression (from the Greek ekzema, which means to boil over). Infancy or childhood is usually where about 80% of disease cases begin, with the remaining instances evolving in adulthood. The disease’s natural course has significant variety, and each person’s path is unique.³⁴ Between the ages of three and six months is when AD most frequently manifests, with about 60% of affected children exhibiting symptoms during the first twelve months.³⁵

Skin burns

According to estimates from the World Health Organization (WHO), burns cause around 11 million injuries and 180,000 deaths annually. Burns occur when heat, chemicals, electrical currents, or other factors cause tissue damage. Burns mostly affect the skin, though they can also impact deeper systems like muscles or bones. The main defenses against the weather, pathogens, evaporation, and heat loss are lost when the skin is burned. The patient’s condition, the stage of the burn, and its cause must all be taken into consideration while deciding on the best course of therapy and effective treatment of burn patients requires interdisciplinary cooperation and personalization. In this comprehensive review, we have collated and analyzed the available treatment options, with a focus on recent advancements in topical medicines, wound cleansing, dressings, skin grafting, nutrition, pain, and scar tissue management as shown in Fig 1.³⁶

Figure 1: Skin diseases Click here to view Figure

Herbs used in treatment of different skin conditions

Aloevera

There have been claims that aloe vera gel can shield the skin from radiation harm.³⁷ Metallothionein, an antioxidant protein that scavenges hydroxyl radicals and protects the skin’s superoxide dismutase and glutathione peroxidase from being suppressed, is produced in the skin after aloe vera gel is administered. Its precise function is unknown. It stops UV-induced suppression of delayed type hypersensitivity by lowering the synthesis and release of immunosuppressive cytokines—like interleukin-10 (IL-10)—derived from skin keratinocytes.³⁸ The goal of the current study was to create and evaluate a polyherbal gel that uses extracts from Vigna radiata and Aloe barbadensis to treat acne, a skin disorder that causes clogged hair follicles and sebaceous glands, which causes skin irritation and redness. Aloe barbadensis pulp was collected, mixed with Vigna radiata extract, and then shaped into a gel using Carbopol 940, triethanolamine, and propylene glycol as the gelling agent, viscosity modifier, and pH modifier, respectively. The antibacterial properties of the gel were evaluated against Staphylococcus aureus, Escherichia coli, and Candida albicans. Antimicrobial medications such as gentamycin and fluconazole met the requirements. The suggested formulation showed a promising zone of inhibition and the gel’s physicochemical properties were evaluated in more detail. Together with Aloe barbadensis’s skin-benefitting properties, the combination had a promising effect on acne.³⁹

The mechanism of Aloe vera involves improving skin health through compounds like acemannan, glucomannans, and gibberellins, which promote collagen synthesis, hydration, and anti-inflammatory effects. These constituents aid in wound healing, reduce irritation, and enhance skin regeneration.

Turmeric

A staple in Asian cuisine and culture is turmeric (Curcuma longa L.). Since ancient times, it has been utilized in conventional medicine. Many health benefits have been attributed to it. In preclinical and clinical trials, the most physiologically active curcuminoid found in turmeric, curcumin, is being studied for its potential to prevent and treat disease. Its actions are antibacterial, anti-inflammatory, anti-tumor, anti-proliferative, and antioxidant. We check the chemical makeup of this plant, its cultural significance in Indian skin care, and its dermatological use.⁴⁰ The C. longa rhizome’s ethanolic extract was utilized to make a gel formulation in several concentrations (1, 2, 3, and 4%). The gel’s topical anti-inflammatory properties were also evaluated. The gel was made with ethanol, propylene glycol 400, methyl paraben, propyl paraben, tri-ethanolamine, ethylenediaminetetraacetic acid, C. longa extract, Carbopol® 940 (1% w/v), and the necessary amount of distilled water. The generated formulations were assessed for their physical properties, pH, spreadability, and ability to irritate skin in order to detect toxicity or negative effects. The findings showed that the gel compositions’ homogeneity and appearance were satisfactory.⁴¹

Mechanism of turmeric involves   benefiting of the skin mainly through curcumin, which has strong anti-inflammatory, antioxidant, and antimicrobial properties. Curcumin inhibits inflammatory mediators and oxidative stress, aiding in acne control, wound healing, and improving skin tone.

Neem

The big, indigenous Indian tree Azadirachta indica, sometimes known as neem, has long been used for its many benefits, chief among them the treatment of skin conditions and its “herbicidal” characteristics. The presence of active secondary metabolites having biological effects, primarily limonoids and tetranortriterpenoids like azadirachtin, makes its bark, leaves, seeds, fruits, and flowers useful in medicine. Therefore, A. indica was investigated as a biopesticide and as an anticancer, antibacterial, anti-inflammatory, and chemopreventive agent. Additionally, in the culture of A. indica, differentiated cell tissue has been shown to generate active metabolites for various uses. On the other hand, very little research has been done on its possible application in cosmetics. For example, the majority of research described the antibacterial qualities in relation to personal hygiene, dandruff, and acne. In order to help researchers and businesspeople choose A. indica derivatives as innovative cosmetic ingredients, we have compiled here not only the most popular cosmetic claims to treat acne but also the mitigation of other skin conditions linked to inflammatory and oxidative processes in recent in vivo studies and patents.⁴² Three medicinal plants with strong anti-inflammatory properties—Cynodon dactylon (L.) Pers., Cassia tora Linn., and Cassia alata Linn.—were chosen for the current investigation and created into polyherbal gels and the gels were made from dried methanolic extracts of Cassia tora Linn, Cassia alata Linn, and Cynodon dactylon (L.) Pers. Polyherbal gel compositions were tested for pH, viscosity, homogeneity, spreadability, and skin irritation. Formalin and carrageenan-induced rat paw edema was utilized to investigate the anti-inflammatory properties. The individual and polyherbal gel of Cynodon dactylon (L.) Pers., Cassia alata Linn., and Cassia tora Linn. were found to have anti-inflammatory qualities in both acute and chronic contexts. When compared to separate gels, polyherbal gel also shown a synergistic effect that may be helpful in treating local inflammation. The solid, jelly-like material known as herbal gel can be prepared in a variety of ways, from soft and weak to firm and robust. It is applied topically for a number of reasons, including antimicrobials, antiseptics, and protectants. Rubia cordifolia, Berberis aristata, Curcuma longa, and Azadirachta indica were combined to create the herbal gel. The antibacterial activity was tested on Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. The most significant effect on hand bacteria, or Staphylococcus aureus, was observed with the herbal gel. Further research is necessary to completely comprehend the mechanism of action and to develop a formulation that can be helpful in the health sector.⁴³

Neem exerts its effects on the skin through compounds like nimbidin, nimbin, and azadirachtin, which provide antibacterial, antifungal, and anti-inflammatory actions. These bioactive constituents help combat acne, reduce skin infections, and promote healing.

Chamomile 

The plant chamomile, which belongs to the Asteraceae family, is native to the Mediterranean and southern Europe and this species has a wide range of secondary metabolites, including as flavonoids, coumarins, sesquiterpenes, and polyacetylenes, and the phytochemical composition varies depending on the growing location. The ethnobotanical knowledge of this plant has led to the widespread usage of chamomile for a variety of health issues worldwide.⁴⁴

Chamomile acts on the skin through compounds like apigenin, chamazulene, and bisabolol, which provide anti-inflammatory, antioxidant, and soothing effects. These constituents help reduce redness, calm irritation, and support skin healing and regeneration.

Rosemary

The medicinal plant known as rosemary, or Rosmarinus officinalis L., is native to the Mediterranean region and is grown all over the world. In addition to its medicinal uses, it’s frequently used as a condiment and food preservative. A variety of pharmacological actions, including anti-inflammatory, antioxidant, antibacterial, antiproliferative, anticancer, and protective, inhibitory, and attenuating properties, are implemented by the bioactive molecules, or phytocompounds, that make up R. officinalis L. Therefore, this Review included both in vitro and in vivo research that address the therapeutic and preventative effects of R. officinalis L. on certain physiological illnesses produced by chemical, biological, or biochemical agents. Thus, the methods, processes, findings, and conclusions were explained. The primary goal of this study was to demonstrate that plant-based products may be used in place of currently available medications.⁴⁵

Rosemary benefits the skin through compounds like rosmarinic acid, carnosic acid, and ursolic acid, which exhibit antioxidant, anti-inflammatory, and antimicrobial properties. These bioactives help protect against oxidative damage, reduce acne, and improve skin tone and elasticity.

Indian privet

Leaves were extracted aqueously to create the gel. Good medication release was demonstrated by the herbal gel formulation. The medication concentration, pH, viscosity, diffusion, spreadability, and extrudability of the herbal gel were all assessed. The gel’s medication release was satisfactory. The gel was translucent and uniform. The pH of the gel was within the acceptable range and correlated with the skin’s pH. Both spreading and removing the gel are simple processes. The gel was thick, viscous, and extrudable enough to come out of the container. Every evaluation parameter was used and stayed within the parameters Vitex negundo gel was created and assessed in response to the growing market for herbal treatments and the paucity of research that conducted an aqueous extraction of Vitex negundo leaves.⁴⁶

Indian privet (Clerodendrum inerme) mechanism involves that, it acts on the skin through compounds like flavonoids, saponins, and phenolic acids, which provide anti-inflammatory, antioxidant, and antimicrobial effects. These constituents help soothe irritated skin, promote healing, and protect against infections and oxidative stress.

In light of the fact that topical herbal gels, which combine the therapeutic advantages of natural substances with cutting-edge formulation techniques, have become viable skin care solutions. This review assesses herbal gel formation procedures as well as physicochemical, biological, and clinical evaluation techniques. Important herbal components including tea tree oil, glycerine, turmeric, and aloe vera are examined for their qualities and effectiveness in treating different skin disorders. The review highlights how crucial it is to choose the right gelling agents, adjust pH levels, and carry out stability testing in order to guarantee the efficacy and safety of the product. Clinical studies demonstrate how well these gels work to treat ailments like irritation, inflammation, and acne. The purpose of this research is to present a thorough analysis of topical herbal gels and encourage their incorporation into standard dermatological therapy. The prepared gel’s physical characteristics, pH, spreading ease, thickness (viscosity), and general smoothness were all evaluated. Additionally, it was tested on human volunteers to see how well it worked on cracked heels and on rabbits to see if it caused skin irritation. Stability studies were carried out in compliance with ICH regulations and the gel was smooth, distributed smoothly, and had a nice appearance, according to the results. Between 4200 and 4500 centipoises was its thickness. The animals showed no signs of skin discomfort. During a two-week research, the gel also effectively decreased the quantity and size of pimples and aided in the healing of human volunteers’ cracked heels. It was also effective in curing skin cracks, rashes, and edema. It can be just as significant for the skin.⁴⁷ Various properties like anti-inflammatory, anti-aging, anti-ulcer, anti-bacterial, antiseptic, etc., that are found in herbs were shown in table 1.

Table 1: Properties of herbs

Conclusion 

According to the review’s conclusion, polyherbs can be utilized to create gels with a variety of qualities, including antibacterial, anti-inflammatory, antiseptic, anti-tumor, and analgesic effects. Neem, turmeric, aloe vera, chickweed, chamomile, rosemary, oregano oil, tea tree oil, and thyme are among the herbs that can be used to make gel. Every element in the composition has a maximal therapeutic impact and little to no negative effects on the body. because every ingredient is a natural product that aids in and promotes the healing of wounds. It was discovered that the herbal gel was a natural product that could be used for a long time after being examined from all angles. The created formulation had antibacterial and anti-inflammatory properties, was more consistent, had greater spreadability, and showed no evidence of phase separation. 

Acknowledgement

We thank management of CMR College of pharmacy for their continuous   support and encouragement. 

Funding Sources

The author(s) received no financial support for the research, authorship, and/or publication of this article.

Conflict of Interest

The authors do not have any conflict of interest.

Data Availability Statement-

This statement does not apply to this article.

Ethics Statement

This research did not involve human participants, animal subjects, or any material that requires ethical approval.

Informed Consent Statement

This study did not involve human participants, and therefore, informed consent was not required.

Clinical Trial Registration

This research does not involve any clinical trials.

Permission to reproduce material from other sources

Not Applicable 

Author Contributions

Vangala Tulasi Iswariya: Conceptualization, Methodology

Anees Mohammed: Writing – Original Draft.

Kota Sanihitha: Analysis, Writing – Review & Editing.

Neerudi Sai Kiran: Administration, supervision.

Tadikonda Rama Rao: Visualization, Supervision,

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The prefix “nano” originates from the Greek word “nanos,” which means “dwarf.” Nanotechnology is the manipulation and application of artificial particles or systems that have at least one dimension smaller than 100 nanometers (nm). A structure that has at least one size (length, height, and breadth) smaller than 100 nanometers (10^-7 meters) and as huge as a virus particle is designated as a nanomaterial. They are categorized depending on their size, dimensions (0, 1, 2, 3D), composition, shape (nanoparticle, nanotube, nanostick, nanofiber etc.), source (natural, synthetic), and content (carbon-derived, composite-based organic-based, etc.). The most often used types of nanomaterials are nanoparticles, which have three dimensions that are all equal and smaller than 100 nanometers.¹ “Nanoparticles” that are found naturally or as results of other operations, such as carbon black, fire smoke, or welding fumes; rather, the word refers solely to manmade particles (such as nanotubes made of carbon, iron oxides, fullerenes, etc.² Humans are exposed to nanoparticles both directly and indirectly as a result of their growing use in commercial applications such as semiconductor components, skincare products, water filtration, fillers, opacifiers, catalysts, and microelectronics.³ In order to target certain cells, biomolecules like DNA, proteins, and monoclonal antibodies are frequently purposefully coated onto nanomaterials for imaging and drug delivery.⁴When it comes to their use in commercial items, industrial processes, and biology, nanomaterials have had a significant impact. This technology, known as nanotechnology, has the potential to bring about a new revolution in society.⁵

Therapeutic applications of Nanoparticles a relatively recent subject of the health sciences, is one of the emerging fields. Consequently, a number of nanoparticles are being investigated or employed in several clinical domains, such as therapy, smart ways to deliver drugs, surgery, medical implants, treatment of illnesses or cancers, and gene delivery as mentioned in the Figure 1.

Gene therapy

The intention of gene therapy, a subject that is extensively researched, is to fix malfunctioning genes in order to prevent and treat genetic illnesses.

Antibacterial activity

Multiple silver nanoparticles have been demonstrated to possess antibacterial activity; these can also be used alongside with other drugs for reducing antibiotic resistance.

ZnO(Zinc oxide) nanoparticles may be regarded as a useful adjuvant in ciprofloxacin combination therapy due to their possible synergistic action with the antibiotic.⁶

Cancer diagnosis and treatment

Carbon nanotubes are employed in the identification of biomarkers and the disclosure of DNA alterations.⁷ Quantum dots are used with magnetic resonance imaging to more precisely reveal tumor location.⁸

Neurological impairment therapy

Applications of nanotechnologies show promise in the management of the neurological disorder and the repair of damaged axons. Utilizing nanoparticles with a strong affinity for circulating amyloid-β with an emphasis on protecting neuronal tissue.⁹

Orthopedic implants

Collagen, hyaluronic acid, chitosan, and titanium alloys are examples of natural and synthetic polymers that are frequently utilized as nanomaterials in bone and cartilage tissue engineering.¹⁰

Therapies for skin health

Novel nanoscale materials that have been created and produced to solve current wound care issues. Drug delivery, growth factor supplements, hydrogels, biodendrimers, electrospun nanofibers, and other polymer therapeutic conjugates have all been employed as skin substitutes in wound healing procedures.¹¹

Nanotoxicity

A novel area of toxicology called nanotoxicology was suggested to fill in the information gaps and to explicitly address the potential negative health effects of nanomaterials. Nanomaterials are being used more and more in industrial applications, consumer goods, and medical devices as nanotechnology develops quickly. Their distinct characteristics, like size, surface area, and reactivity, can, nevertheless, result in unanticipated toxicological effects that are not found with bulk materials. Nanotoxicology encompasses the domains of exposure pathways, molecular determinants, physicochemical factors, biodistribution & genotoxicity.¹²

Figure 1: Therapeutic applications of Nanoparticles       Click here to view Figure

Physicochemical properties of nanoparticles

 Nanoparticles possess unique physicochemical properties that distinguish them from their bulk counterparts, such as extremely small size and a high surface area-to-volume ratio as shown in the Figure 2. These features grant them enhanced reactivity, improved cellular uptake. However, the same properties that provide functional advantages can also introduce novel mechanisms of toxicity. Their small size allows nanoparticles to interact with cellular structures at the molecular level, potentially disrupting normal biological functions. High surface reactivity can lead to the generation of reactive oxygen species (ROS), causing oxidative stress, inflammation, and cellular damage. Different shapes and morphologies, such as rods or fibers, may lead to mechanical damage or hinder cellular clearance, prolonging exposure. Thus, nanoparticle-induced toxicity arises not from a single factor but from a complex interplay of size, surface area, composition, shape, and other physicochemical traits.

Particle size and surface area

Materials’ surface area seems to grow exponentially in proportion to their volume as they get smaller, which raises the nanomaterial’s surface reactivity to its surroundings and to itself. Notably, particle size and surface area affect how the system responds to, distributes, and eliminates the materials.¹⁴ To evaluate the in vitro cytotoxicity of NPs of different sizes, several groups have employed a variety of cell types, incubation conditions, and duration of exposure.^15,16 One of the primary reasons for the toxicity of ENMs(Engineered Nanomaterials) in vivo is the generation of reactive oxygen species through the creation of free radicals. Dimensions has significance in this process because, as many writers have observed, the smaller the object, the more likely it is to generate ROS(Reactive oxygen species).¹⁷ Numerous investigations utilizing a range of nanoparticle classes demonstrated that surface area plays a key role in exhibiting hazardous effects.¹⁸ While NPs larger than 50 nm do not spread to other tissues and are readily absorbed by RES(Reticulo endothelial system), NPs smaller than 50 nm have been found to traverse rapidly to almost all tissues and produce possibly harmful symptoms in a variety of tissues.¹⁹

Effect of Particle Shape

Numerous nanoparticles, such as silica, allotropies, carbon nanotubes, gold, nickel and titanium nanomaterials have been shown to have shape-dependent toxicity.^20-23 In contrast to nanoparticles that are fibre-like or rod-shaped nanoparticles, spherical nanoparticles have been found to undergo endocytosis more easily and quickly.²⁴Likewise, was demonstrated that the absorption of gold nanorods is not as quick as spherical nanospheres and that it peaks when the aspect ratio is close to unity.²⁵

Effect of Surface Charge

Surface charge has a substantial impact on nanoparticle toxicity as well. Because of their improved opsonization by the plasma proteins, positively charged nanoparticles exhibit notable cellular absorption in contrast to negatively charged and neutral nanoparticles. Additionally, they have been demonstrated to cause platelet aggregation and hemolysis,²⁶ besides it was also noted that the surface charge of nanoparticles modifies the transmembrane permeability and blood-brain barrier integrity.

Effect of Aggregation and Concentration

Research findings demonstrated that well-dispersed carbon nanotubes had cytotoxic effects when contrasted with those of conventionally purified rope-like agglomerated carbon nanotubes and asbestos as a reference.²⁷

Figure 2: Physicochemical properties affecting metal based nanoparticle toxicity-Different sizes and shapes.Click here to view Figure

Created in https://BioRender.com

Routes of exposures

One definition of “exposure” is the potential for an agent or substance from the outside environment, such as a nanoparticle (NP), to enter the body as shown in the Figure 3. Furthermore, the potential toxicity is determined by the exposure route.²⁸

Inhalation Exposures

Exposures through inhalation are known to occur largely in professional environments, have been the focus of a large portion of exposure and toxicological research efforts and measurements using NPs to date. In fact, according to research on animals, the majority of the harmful effects of inhaled NPs or particles affect the cellular components in the anatomical compartment of the respiratory tract, namely at the portal entrance sites. The lungs capillaries are where carbon dioxide and oxygen exchange gases. Although the possibility is thought to be minimal or infrequent, it is possible that inhaled NPs could eventually enter the systemic circulation through this pathway.²⁹

Dermal Exposure

In essence, the skin is made up of two main sections: the larger, known as the epidermis, and the dermal component, which is located underneath the epidermis and has a restricted supply of blood vessels. Even after prolonged application, Lademann noted that no particles were seen in the stratum corneum’s deeper levels.³⁰ Exposures via the gastrointestinal tract happen after consuming food or after swallowing and pulmonary clearance of inhaled nanoparticles.³¹

Oral or Ingestion Exposures

More precisely, food coloring, supplements, and flavor enhancers are a few examples of ingested component sources.³² Depending on the kind of nanoparticle and its characteristics, the possibility that it will pass through the digestive system varies; many are simply expelled without entering the body. According to some researchers, oral exposure to NPs may cause absorption through the gut-associated lymphoid tissue’s Peyer’s patches’ epithelial cells. On an average, micro-copper particles do not cause significant harm to the liver, kidney or spleen of experimental mice but nanoparticles do. The findings point to a gender-dependent aspect of nanotoxicity. The findings point to a gender-dependent aspect of nanotoxicity. ³³  According to recent research,³⁴ NPs administered orally may be absorbed through the lymph nodes and travel throughout the gastrointestinal tract.

Figure 3: Different routes of exposure of nanoparticles.Click here to view Figure

Created in https://BioRender.com

Biodistribution of Nanoparticles

Physical clearance processes (such as epithelial endocytosis, mucociliary movement, lymphatic drainage, interstitial translocation& blood circulation,) and chemical clearance processes (such as leaching, protein binding & dissolution) are implicated in the studies conducted thus far. Certain nanoparticles may build up in the liver as a result of first-pass metabolism.³⁵ Nanoparticles are transferred to the bone marrow, colon, lungs, spleen, liver and lymphatics following intravenous injection.³⁶ Differences in rate of clearance & confinement of nanoparticles by macrophages are a result of differential opsonization.³⁷ Opsonization events must be suppressed at certain locations or anatomical compartments in order to enhance the non-invasive retention of nanoparticles in the bloodstream. For instance, a poly(ethylene) glycol (PEG) coating would make hydrophobic particles more hydrophilic, lengthening the time it takes for them to circulate throughout the body.³⁸

Genotoxicity

 The genotoxic potential of NM exposure is a major worry. The ability of physical or chemical substances to change genetic information is known as genotoxicity. Genotoxic events can result in permanent alterations (mutations) in the quantity or structure of a cell’s genetic material, or they might be temporary (repairable harm). A genetic condition may result from a mutation in a germ cell that is passed down to the following generation. Cancer in somatic cells can result from a mutation in a crucial gene. Therefore, all mutagens are thought to have the potential to cause cancer.³⁹ Several studies have been conducted on the genotoxicity of NMs and the processes that result in temporary or permanent genetic alterations. According to recent research, there are two primary (direct or indirect) and secondary genotoxicity mechanisms that might cause NM genotoxicity.^40,41Physical contact between NMs and DNA in the nucleus is necessary for primary direct genotoxicity, which is brought on by direct interaction between NMs and the genome. Large DNA malformations, chromosomal damage, and DNA breaks and other diseases could result from this interaction. Physical contact between NMs and DNA in the nucleus is necessary for primary direct genotoxicity, which is brought on by direct interaction between NMs and the genome. Large DNA malformations, chromosomal damage, and DNA breaks and other diseases could result from this interaction. The main indirect mechanism is caused by reactive oxygen species (ROS) triggered by NMs or by toxic ions released during NM dissolution and intracellular ROS formation through the Fenton-type reaction.^42,43 The primary mechanism of NM genotoxicity is thought to be secondary genotoxicity. ROS generated by inflammatory cells mediate it. By activating phagocytes, NMs can generate an oxidative burst. This inflammation, which aids in clearance, is the body’s first line of defense against the invasion of microbes or other objects.⁴⁴A range of tests covering DNA damage, gene mutations, and chromosomal damage as sensitive genotoxicity endpoints are included in the category of genotoxicity biomarkers. The number of NMs produced is constantly increasing despite the lack of information and limited data regarding NM safety. For this reason, attempts have been made to comprehend how NMs work and to create or modify approved OECD testing procedures for chemicals for use on NMs in a number of previous and ongoing European projects, including NanoGENOTOX, NanoTEST, NanoReg, RiskGONE, HISENTS, RiskGONE, and others.⁴⁵ There are issues with the mode of action (MoA), specificity and data of regulatory In vitro genotoxicity testing.⁴⁶

Methods for assessing toxicity of nanomaterials

In Vitro Methods

They are essential to nanotoxicology because they offer a controlled setting for examining interactions of nanoparticles with biological systems, including human cells, tissues, and other model species. In vitro toxicity studies often look at the following parameters: oxidative stress, inflammatory alterations, apoptosis, DNA mutation/damage and cytotoxicity. Inflammatory alterations are an additional parameter that can be assessed using in vitro testing. Inflammatory indicators can be found using the enzyme-linked immunosorbent test (ELISA).⁴⁷Moreover, in vitro tests can also be used to assess cytotoxicity or necrosis. Assays for cytotoxicity include lactate dehydrogenase (LDH), clonogenic assays, Trypan blue assays, Tetrazolium salts assays, and Alamar blue assays.⁴⁸

Proliferation Assay

One important factor in the growth and progression of tumors is cell proliferation, which can also represent the surrounding cells’ compensating reaction to necrosis or apoptosis. Currently, there are several well-established techniques for measuring cell proliferation each with unique limitations and specificity. Currently used techniques include flow cytometry, immunohistochemistry, and histochemistry. Histochemical techniques include the absorption of the DNA analog bromodeoxyuridine (BrdU), incorporation of tritiated thymidine ([3H] thymidine), and direct detection of mitosis by staining for DNA content⁴⁸. In immunohistochemistry, Ki-67 and antibodies that identify proliferating cell nuclear antigens (PCNAs) are now of interest.⁴⁹

Apoptosis Assay

This assay assays the activity of caspase-3 and caspase-7 using luminescence. Cell lysis and substrate cleavage by caspase occur when caspase reagent is added to NP-treated cells. Luciferase then generates a luminous signal that is equivalent to the amount of caspase activity that is available.⁵⁰

 Genotoxicity Assays

Chromosomal aberration induction

This method involves assessing how NPs affect the quantity of cells and alterations in the physical appearance of chromosomes. Cells are treated with NPs during the S-phase because they are more responsive during this phase. Colcemid or colchicine is then administered at predetermined intervals. Trypsinated cells are counted after chromosomes are processed and stained with Giemsa stain for microscopic evaluation of aberration (chromatid exchange, chromatid breaks, chromosomal breaks, chromosome fragmentation)⁵¹ It is employed to measure and evaluate cellular DNA damage. After cells have been embedded in a thin agarose gel and put on a microscope slide, the cells are lysed to extract the cellular proteins. In neutral or alkaline circumstances, allowing the DNA to uncoil before being subjected to electrophoresis, which allows the pieces of DNA to detach from the nucleus. The amount of fluorescence in the head and tail length is determined by staining with either propidium iodide or ethidium bromide. The amount of damaged DNA is directly equivalent to the amount of DNA released from the comet’s head.^52,53

Measurement of oxidized guanine bases

Single base changes within a specific gene can be detected by assaying any one of the many oxidized guanine bases, including 7,8-dihydro-oxodeoxyguanine (oxo-dG) and 8-hydroxydeoxyguanosine (8-OHdG). These bases alterations are often caused by oxidative damage and are quantified by HPLC or immunohistochemistry.⁵⁴

In Vivo Methods

Using animal models, such as mice and rats. In situ toxicity is typically assessed. The methods used to evaluate in vivo toxicity include histopathology, biological distribution, clearance, hemophilia, and plasma chemistry. Biodistribution studies examine how nanoparticles enter organs or tissues. Nanoparticles can be detected in living and expired organisms using radiolabels.⁵⁵ The liver, kidneys, heart, brain and spleen are among the tissues exposed to nanoparticles, according to histopathology research.⁵⁶ Examining alterations in cell type and serum chemistry following nanoparticle exposure is an additional method of determining in vivo toxicity.⁵⁷

Table 1: An overview of the research on nanomaterial toxicity tests

Novel Technologies Employed for Evaluation of Toxicity

Ever since the development of nanotechnology, a growing number of chemicals have entered the environment, necessitating the collection of data regarding their toxicity. Traditional toxicity assessment using animal models is often not practical in these circumstances due to its poor capacity, time-consuming nature, high expense, and limited evaluation of endpoints. The authors claim that this method, in conjunction with functional toxicogenomics, can be crucial in determining the key biological elements and pathways implicated in the toxicity response.⁷⁰

Genome arrays have been employed to evaluate the impact of nanoparticles. The expression of many genes linked to neurodegenerative illness, immune cell function and motor neuron disorders was altered by the inhaled silver nanoparticles, suggesting possible immunotoxicity and neurotoxicity.⁷¹The use of high-throughput screening (HTS) techniques to check how silver nanoparticles affect bacterial cells is said to aid in tracking the ecological consequences of nanoparticles.⁷² Additionally, HTS might make it possible to create models that forecast how nanoparticles will behave in biological systems. HTS’s objective is to support the safe development of nanomaterials by using quick, automated screening techniques to produce comprehensive, comparable toxicity data for thousands of distinct nanomaterials.⁷³

Computational Models

Computational methods made possible by the development of AI   and ML allow nanomedicine to investigate safety issues effectively and economically. The safety assessment at the early phases of drug research, which integrates data at several levels and produces reliable conclusions, has a detrimental impact on the medication’s failure in the drug development process. Understanding the limitations, science and opportunities that underpin this application is crucial to getting the most out of it. Additionally, computational approaches consider the pharmacokinetic elements in addition to the ligand-receptor docking concept in order to illustrate the results.⁷⁴

Quantitative Structure-Activity Relationship (QSAR) Modeling

Routine nanotoxicity investigations use less time, money, and resources thanks to computational technologies like nano-QSAR models QSAR and (at the nanoscale). Pharmacokinetic and pharmacodynamic data are primarily correlated with in vivo application scenarios using these models.⁷⁴ QSAR methods use a compound’s physicochemical characteristics to predict its biological activity ⁽⁷⁵⁾. Later, in 1988, Cramer and associates created a different method known as 3D-QSAR, which takes into account the interactions, activity& spatial structure of molecules.⁷⁵ One-dimensional (1D), two-dimensional (2D), and three-dimensional (3D) techniques are all covered by Nano-QSAR, making it a very universal model⁷⁶. The low-energy conformations that docked into the ADME model were used to build the 3D nano-QSAR. Atoms in a crystal are more likely to overlap their orbital energy and divide if they are close to one another. Conduction band overlap suggests that NMs are cytotoxic or have other disruptive effects.⁷⁶

Stem Cells as a Novel Approach to Predict Toxicity

All of the biological cell can be developed from pluripotent stem cells. The majority of the precise NMs toxicity is still unknown because of the use of imprecise, oversimplified techniques, such as 2D cell lines, in compound testing and validation. Recent stem cell research has focused more on ESCs and iPSCs.⁷⁷ ESC is produced from an embryo’s undifferentiated inner mass cells, whereas iPSCs are derived from somatic cells that have been reprogrammed to function as ESCs by activating genes or forcing the expression of reprogramming genes such Klf4, Oct4 and Sox2.⁷⁸

Omics Technologies

“As a whole” is what the suffix “omics” stands for, and it encompasses proteomics, metabolomics, transcriptomics, epigenomics, and genomics. Adaptive responses can be identified using these techniques. Systems toxicology encompasses transcriptomics, proteomics, metabolomics, genomics, and epigenomics (miRNomics and DNA alterations). Using recombinant DNA, DNA sequencing, and bioinformatics to examine the structure and function of the genome, genomics studies genes and their roles. These technologies’ main benefits include the ability to forecast toxicity at low exposure levels to nanoparticles, which can stress cells without causing toxicity.⁷⁹ NPs cause this pattern and affect a number of biological functions, including membrane integrity, inflammation, apoptosis, and proliferation.⁸⁰

Conclusion

Nanotoxicology is a rapidly expanding field focused on assessing the risks of nanomaterials used in industries like healthcare and consumer products. While nanotechnology offers significant therapeutic potential, such as in drug delivery and cancer treatment, the unique properties of nanoparticles—such as size, surface area, shape, and charge—can lead to unexpected toxicological effects. Understanding the mechanisms of nanoparticle toxicity, including genotoxicity, inflammation, and oxidative stress, is critical for ensuring their safe use. The biodistribution, clearance, and exposure routes of nanoparticles (inhalation, dermal contact, and ingestion) significantly affect their health impacts. Despite progress, many gaps remain in understanding nanomaterial safety, and more research is needed to establish comprehensive safety standards. Advancements in novel testing methods, including high-throughput screening, computational models, and stem cell-based assays, are essential to assess the long-term effects of nanoparticles and guide their safe development. Both in vitro and in vivo methods are crucial for evaluating the biological effects of nanoparticles, with assays focused on cytotoxicity, oxidative stress, genotoxicity, and apoptosis being commonly used. Emerging technologies like high-throughput screening, computational models, and omics technologies offer efficient, cost-effective ways to predict nanoparticle behaviour and toxicity. These tools enhance our ability to assess the molecular impacts of nanomaterials, identify adaptive responses, and predict low-exposure toxicity. As nanomaterials continue to evolve, integrating these advanced methods is vital for ensuring the safe development and regulation of nanomaterials, protecting both human health and the environment.

Acknowledgement

The Author would like to Profoundly Grateful to the Principal and Management of Vignan Institute of Pharmaceutical Technology (Autonomous). Visakhapatnam.

Funding Sources

The author(s) received no financial support for the research, authorship, and/or publication
of this article.

Conflict of Interest

The authors do not have any conflict of interest.

Data Availability Statement

This statement does not apply to this article.

Ethics Statement

This research did not involve human participants, animal subjects, or any material that
requires ethical approval.

Informed Consent Statement

This study did not involve human participants, and therefore, informed consent was not
required.

Clinical Trial Registration

This research does not involve any clinical trials.

Permission to reproduce material from other sources

Not Applicable

Author Contributions

Galanki Vasantha : Conceptualization, Supervision, Methodology, Writing- Review & Editing.

Rangala Mohini: Writing – Original Draft, Data Collection, Analysis

Iragavarapu Tejolahari: Analysis  and Data Collection,

Gajula Niharika: Data Collection and Analysis

Allampalli Likhita: Data Collection and Analysis 

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Climate change and associated disease emergence is one of the most important global issues of the twenty-first century with a cornucopia of impacts.¹ Because infectious diseases have the potential to cause widespread morbidity and mortality, their burgeoning risks associated with extreme weather events, changing precipitation patterns and rising global temperatures have changed ecosystems and the dynamics of pathogen-host interactions.² All these has made it easier for infectious diseases to arise, resurface and spread geographically at an alarming rate. The relationship between climate change and public health becomes increasingly significant as the planet warms and the weather shifts, particularly with regard to infectious diseases. More incidences of diseases result from improved conditions for pathogens and insects like mosquitoes and ticks brought on by rising global temperatures.^3,4 Moreover, natural disasters and extreme weather can damage healthcare systems, increasing the likelihood of breakouts in communities. It is evident from examining historical trends, contemporary illness patterns and projections for the future that addressing how infectious diseases are impacted by climate change is essential for international health safety and response strategies. It should be noted that changes in the frequency and distribution of vectors and diseases to climate change are also influenced by changes in land use,⁵ the abundance of reservoir hosts⁶ and various control methods adopted.⁷ Furthermore, even while scientific methods to differentiate between human-induced change and natural climate variability are developing, it may be challenging. Notwithstanding these complications, it is evident that the pathogens, vectors, and reservoir hosts that make up vectorborne disease systems are extremely sensitive to the diverse environments in which they live, and that observed variations in the prevalence of vectorborne diseases at particular sites are frequently linked to corresponding shifts in the local climate. Thus, the paper is an attempt to investigate the intricate relationships between climate change and the emergence and spread of infectious diseases, aiming to elucidate how environmental factors, disease vectors and human health outcomes interact amid changing climatic conditions.

Climate change and disease dynamics

The dynamics of infectious diseases are being drastically changed by climate change, which is also changing their prevalence, patterns and effects on the health of humans and animals.⁸ In another study, climate change is a health emergency not only for humans and animals but also for the environment.⁹ The emergence and spread of numerous infections and their vectors are being facilitated by factors such as rising global temperatures, shifting precipitation patterns and an increase in the frequency of extreme weather events. Once restricted to particular areas, diseases are now spreading geographically, presenting new threats to people that were previously untouched. Warmer temperatures and longer mosquito and tick breeding seasons have resulted in notable changes in the spread of vector-borne diseases. Likewise, waterborne diseases also flourish in areas affected by flooding and inadequate sanitation, all of which are made worse by climate change. Furthermore, as evidenced by the COVID-19 and Nipah virus outbreaks, habitat loss and deforestation are causing changes in ecosystems and biodiversity, which in turn are increasing human susceptibility to zoonotic diseases.¹⁰ Existing public health systems are put to the test by these changing disease patterns, which calls for improved methods of response, prevention and surveillance. In a time of incessant environmental change, it is essential to realize the complex relationships between disease dynamics and climate change in order to mitigate their effects and protect global health.

Temperature and pathogen survival

Many pathogens and vectors reproduce and spread more quickly in warmer climates. Warmer temperatures, for example, prolong the mosquito breeding season, which increases the spread of vector-borne illnesses like dengue, malaria, and the Zika virus. In temperate regions, diseases traditionally confined to tropical areas are now spreading due to warmer winters and longer summers.¹¹

Precipitation and waterborne diseases

Waterborne illnesses like leptospirosis and cholera are more likely to occur when rainfall patterns change.¹² Droughts can concentrate water sources, which encourages the spread of diseases in communities that are water-stressed, while heavy rainfall and flooding can taint drinking water sources with pathogens.

Extreme weather events

Hurricanes, cyclones and heatwaves disrupt infrastructure, displace populations, and create conditions conducive to disease outbreaks.¹³ For example, the aftermath of Hurricane Maria in Puerto Rico saw a surge in leptospirosis cases due to contaminated floodwaters.¹⁴

Ecosystem disruption

Disease vectors and reservoirs proliferate as a result of ecological changes brought on by climate change, which also disturbs predator-prey relationships and affects biodiversity.¹⁵ For instance, habitat loss and deforestation frequently increase human-wildlife contact, raising the possibility of zoonotic spillovers, such as those caused by COVID-19 and the Nipah virus.¹⁶

Emerging and resurging diseases

Diseases that are on the rise pose serious threats to public health systems, economies and society worldwide with far-reaching ramifications. Diseases classified as emerging occur when they first manifest in a community or when their incidence or geographic range rapidly expands.¹⁷ Conversely, resurging diseases are those that were formerly under control but are now resurfacing as a result of shifting biological, social and environmental factors.¹⁸ Numerous interrelated variables, such as population migration, deforestation, urbanization, globalization, climate change and antibiotic resistance, are responsible for these phenomena.¹⁹ These factors have facilitated the global spread of infectious illnesses, the resurgence of latent infections and the transfer of pathogens from animals to people. The advent of illnesses like COVID-19, SARS, and MERS highlights how zoonotic transmission plays a part in public health emergencies and is frequently connected to changes in land use and close human-wildlife contact.^20-23 Similar to this, resurgences of diseases like dengue, malaria and tuberculosis show how disease control strategies are becoming less effective as a result of variables including medicine resistance, climate variability and vector adaptation.^24-27 As demonstrated by the increased spread of infectious agents through global travel and trade, globalization also makes it easier for viruses to spread quickly.²⁸ These difficulties are made worse by the effects of climate change, which change the habitats and behaviours of vectors and cause diseases like Lyme disease, Zika and chikungunya to spread geographically. Furthermore, the danger of foodborne and waterborne illnesses is increased by urbanization and inadequate sanitation. Additionally, antimicrobial resistance contributes to the return of viral and bacterial infections, making treatment approaches more challenging and raising fatality rates.²⁹ Numerous health risks have emerged and resurfaced as a result of the interaction between infectious illnesses and climate change. In addition to spreading geographically, these illnesses are also changing in intensity and seasonality.

Vector-borne diseases

With millions of cases annually, vector-borne diseases represent serious risks to world health. These illnesses are impacted by a complex interaction of biological, socioeconomic, and environmental factors.³⁰ Diseases have spread geographically and formerly controlled infections have resurfaced as a result of changes in vector habitats and behaviour brought about by climate change, urbanization, deforestation and international travel.³¹ In tropical and subtropical areas, where vulnerabilities are increased by resource constraints and climatic circumstances, the growing burden of vector-borne diseases is especially severe.³² Environmental changes have a direct impact on the dynamics of vector-borne diseases. Vectors’ ability to reproduce and survive is impacted by changing precipitation patterns and rising global temperatures, which allow them to spread their habitats into new areas. As seen by the transcontinental spread of Aedes aegypti mosquitoes and the corresponding spike in dengue and Zika outbreaks, the quick speed of international travel and trade also makes it easier for vectors and diseases to spread.³³

Malaria

Rising temperatures in high-altitude regions and changing rainfall patterns have expanded malaria’s range into previously unaffected areas, particularly in Africa and South America. Recent studies clearly portrayed the environmental drivers, climate change and emergent diseases transmitted by mosquitoes and their vectors in southern Europe.³⁴

Dengue and Zika

Warmer climates have expanded the range of Aedes mosquitoes, the primary vectors for dengue and Zika viruses, into new regions, including parts of Europe and North America. Recent studies highlighted the global expansion and redistribution of Aedes-borne virus transmission risk due to climate change.³⁵

Waterborne diseases

Globally, waterborne diseases are worryingly on the rise due to climate change which include cholera, typhoid, hepatitis A, leptospirosis and giardiasis are spread by tainted water sources. As a catalyst, climate change modifies the environment in ways that increase these diseases’ ability to survive, proliferate and spread.³⁶ Waterborne infections are becoming a greater hazard to public health globally as a result of favourable conditions for their spread and reemergence brought about by rising global temperatures, shifting precipitation patterns and an increase in extreme weather events. Pathogens like Vibrio cholerae multiply more quickly in warm water bodies due to rising temperatures, especially in coastal and estuarine areas. Similarly, dangerous algal blooms, which release toxins that contaminate drinking water and severely affect aquatic ecosystems, are more common in warmer climates. Sewage mixes with drinking water sources as a result of altered precipitation patterns that are marked by heavy rainfall and flooding, overtaxing water and sanitation systems results in developing hotspots for diseases like leptospirosis and dysentery.³⁷ On the other hand, extended droughts decrease the amount of safe water available, which increases the risk of exposure by concentrating pathogens in limited or stagnant water sources. Extreme weather events like hurricanes, cyclones and typhoons exacerbate the recurrence of waterborne diseases by upsetting infrastructure, uprooting populations and exposing communities to contaminated water supplies.³⁸ Particularly, floods increase the likelihood of outbreaks in both rural and urban regions by acting as a medium for the spread of pathogens.³⁹

Cholera

Warmer ocean temperatures have been linked to Vibrio cholerae outbreaks in coastal areas. These bacteria thrive in warmer waters, particularly in regions experiencing increased flooding or water stagnation.

Cryptosporidiosis and Giardiasis

Flooding and inadequate sanitation exacerbate outbreaks of these parasitic infections, especially in low-income and disaster-prone regions. The infectious disease outbreaks in the wake of natural flood disasters along with its global patterns and local implications are well established.⁴⁰

Zoonotic diseases

A major worldwide health concern, especially in light of climate change, is zoonotic diseases, which have a greater impact on pathogens. Through its effects on ecosystems, wildlife behaviour and human-animal interactions, climate change is changing the seasonality, geographic dispersion and modes of transmission of zoonotic diseases, which is contributing to their reappearance.^41,42 It should be noted that global economy, ecosystems and public health are all seriously at danger from these changes. The habitats and behaviours of wildlife and vectors, including ticks and mosquitoes, which are important contributors to the spread of zoonotic diseases, are changing as a result of rising global temperatures. In a similar vein, shifting weather patterns are impacting bat and bird migration and mating habits, which raises the risk of zoonotic infection exposure for humans. These dangers are made worse by extreme weather events like droughts and floods, which uproot livestock and wildlife and increase the likelihood of pathogens spreading to human populations. Additional hotspots for virus introduction and amplification are produced by intensive farming methods, wet markets and the illicit wildlife trade.^43,44 By straining ecosystems and diminishing biodiversity, climate change exacerbates these dynamics by lowering the natural barriers that normally stop the spread of pathogens.⁴⁵ The return of zoonotic diseases emphasizes how closely related human, animal and environmental health.

Lyme Disease

Rising temperatures in North America and Europe have extended the range of Ixodes ticks, the primary vectors of Lyme disease. The disease is termed as “Ticking Bomb” and well-illustrated the impact of climate change on the incidence of the disease.⁴⁶

Hantavirus

Habitat destruction and changes in rodent populations, driven by climate variability have contributed to hantavirus outbreaks. The influence of climatic factors on human hantavirus infections in Latin America and the Caribbean with catastrophic public health impacts.⁴⁷ Several studies reiterated that Hanta virus is an emerging threat for public health.^48,49

Fungal Diseases

Climate change is having an increasing impact on the occurrence and severity of fungal illnesses, which have a burgeoning public health impact globally. As opportunistic diseases, fungi flourish in shifting environmental conditions and the changing climate has made it easier for them to proliferate, spread geographically and adapt.⁵⁰ The prevalence of diseases brought on by fungi like Aspergillus fumigatus, Candida auris, Cryptococcus neoformans and Histoplasma capsulatum has increased dramatically, endangering the health of people, animals and plants. Their capacity to take advantage of compromised ecosystems, pressures brought on by climate change and environmental disruptions caused by humans exacerbates the recurrence of fungal diseases. The evolution and adaptability of fungi are directly impacted by rising global temperatures.⁵¹ Once restricted to particular places, many fungal infections are now spreading to new areas and flourishing in climates that were previously uninhabitable.⁵² Because crops are more susceptible to fungal diseases like Fusarium and Puccinia, these changes also have an impact on agriculture and put food security at risk. Climate change-driven phenomena have also amplified the risk of fungal outbreaks. The resurgence of fungal diseases is further complicated by antimicrobial resistance, which limits treatment options and increases morbidity and mortality rates.⁵³ Many fungi, such as Candida auris, are resistant to first-line antifungal drugs, posing significant challenges for healthcare systems. Moreover, the lack of effective vaccines against fungal infections exacerbates the global burden of these diseases, particularly among immunocompromised individuals.

Coccidioidomycosis (Valley Fever)

Warmer and drier conditions in arid regions of the Americas have led to increased cases of this fungal infection. The emergence of valley fever under a changing climate in the United States was well illustrated.⁵⁴ In another study, the dynamics of coccidioidomycosis in relation to climate in the southwestern United States with significant increase in incidences was also observed.⁵⁵

Candida auris

The emergence of C. auris as a human pathogen under the auspices of climate change was well established.⁵⁶ The emergence of C. auris as a human pathogen have been facilitated by the selective pressure exerted by higher average temperatures brought on by anthropogenic climate change, which favoured strains of the fungi that were acclimated to salinity and higher temperatures, which are similar to those found in the human body.⁵⁷

Vulnerable populations and inequities

Globally, pre-existing social and economic inequities are being exacerbated by climate change, which disproportionately affects vulnerable populations. Vulnerable people are more at risk because of their limited capacity to adapt to the health, economic and environmental effects of a changing climate. Low-income neighbourhoods, native communities, women, children, the elderly and those with pre-existing medical conditions are some of these groups. The unequal distribution of climate change’s effects highlights the connection between structural inequities and environmental deterioration, making it both an environmental disaster and a serious social justice issue.⁵⁸ Natural disasters like hurricanes, floods, droughts and heat waves disproportionately impact marginalized communities. Racism and climate change interaction with disproportionate effects on the lives of minoritised people both within countries and between the Global North and the Global South especially in connection with health and racial justice was also well recorded.^59,60 These populations lack access to resources necessary for disaster recovery and frequently live in high-risk areas, such as urban heat islands or flood-prone zones.

Furthermore, migration and displacement brought on by climate change increase vulnerabilities since displaced people usually face poor living conditions, limited access to services and increased risks of exploitation. Climate change’s economic repercussions make inequality even worse. Vulnerable communities are more likely to rely on climate-sensitive sectors including agriculture, forestry and fishing for their livelihoods. Many people are forced into deeper poverty as a result of extreme weather events and changed rainfall patterns that jeopardize food security and economic stability. Additionally, these groups usually lack the financial and institutional means to implement adaptive measures, which perpetuates cycles of vulnerability and marginalization. Not all infectious diseases are equally affected by climate change. Pre-existing socioeconomic and geographic variables cause vulnerable communities to suffer an unfair burden.

The public health emergency of current global pandemics was well reported⁶¹ with re-emergence of known and future (X) viral pathogens. It is evident that climate change and infectious diseases, especially zoonoses, are tortuously related due to the intricate interplay of ecological disturbances, modified host-pathogen interactions, environmental deterioration and evolving transmission dynamics.⁶² Thus, the world shouldn’t expect that the severity of the sickness in communities who haven’t been exposed to infectious diseases before will be the same as in populations where those diseases have been prevalent for a long time.⁶³ It is expected that the next decade will test humanity’s ability to balance ecological pressures with health security, making proactive science and equitable healthcare access essential tools in the fight against emerging pathogens.

One Health approach

With its emphasis on the interdependence of environmental, animal and human health, the One Health approach provides a thorough framework for tackling the complex issues brought on by climate change.⁶⁴ Climate change increases hazards in all areas of health by upsetting ecosystems, changing the dynamics of illness and intensifying extreme weather events. The One Health paradigm encourages interdisciplinary cooperation to reduce risks, improve resilience and advance global health security because it acknowledges that these changes cannot be handled in a vacuum. The emergence and spread of diseases, especially zoonotic and vector-borne diseases, are significantly impacted by climate change. The habitats of disease-carrying insects like mosquitoes, ticks and rodents are altered by rising temperatures and changing precipitation patterns, which causes them to spread into new areas and expose previously unaffected populations to diseases like dengue and malaria, dengue, Lyme disease and leptospirosis.

Furthermore, human-wildlife interactions are exacerbated by ecosystem degradation brought on by urbanization, deforestation, and climate-related disasters, which makes it easier for viruses to spread from animals to people. To detect and reduce such hazards, the One Health method offers a framework for integrating environmental evaluation, animal monitoring, and epidemiological surveillance. Antimicrobial resistance and food security are also affected by climate change. The One Health framework guide safe food systems, encourage sustainable farming practices and enhance antibiotic usage monitoring to reduce these hazards. Sanitation and water security risks brought on by climate change can also be addressed via the One Health approach. When droughts and flooding degrade water quality, waterborne diseases become more prevalent with unimaginable health impacts. In addition, the One Health concept emphasizes how crucial international cooperation is in mitigating the effects of climate change. It makes it easier to share information, resources and experience in order to improve health systems and put adaptive measures into place by encouraging collaborations across disciplines and national boundaries. In order to increase resilience to climate-related difficulties, this strategy also places a strong emphasis on community involvement, acknowledging the importance of local knowledge and customs. In short, the One Health approach offers a holistic and integrated framework to tackle the complex interplay between climate change and health. By bridging the gaps between human, animal, and environmental health, it provides a pathway for collaborative, sustainable and equitable solutions to mitigate climate risks and enhance global health outcomes. Adopting the One Health approach is not only a necessity but a moral imperative in safeguarding the health of current and future generations in a rapidly changing world.

Conclusion

Climate change is reshaping the landscape of infectious diseases, amplifying existing risks and creating new challenges for global public health. The nexus dynamics of rising temperatures, shifting weather patterns and environmental degradation have enabled the emergence, resurgence and geographic spread of diseases that were once confined to specific regions. Waterborne infections, zoonotic spillovers and vector-borne diseases like dengue and malaria are just a few of the threats to human health and socioeconomic stability brought about by global warming with crippling impacts on life and livelihood. An immediate, multidisciplinary and cooperative effort combining climate research, public health, policymaking and community resilience is needed to address these imminent threats. The challenges posed by climate-driven infectious diseases underscore the need for holistic frameworks, such as the One Health approach, that recognize the interconnectedness of human, animal and environmental health. By addressing the root causes of vulnerability alongside the direct health impacts of climate change, it is possible to build resilience and reduce the global burden of infectious diseases. Ultimately, the fight against climate-induced infectious disease risks is not only a matter of safeguarding health but also of securing a sustainable and equitable future. The convergence of climate action and public health initiatives presents an unprecedented opportunity to protect ecosystems, strengthen global health security and ensure that the feverish future we face is one of innovation, resilience, and hope, rather than crisis and despair. Future epidemiology, climate science and public health research will be greatly aided by this study on the dangers of infectious diseases brought on by climate change. Emerging infections and changing routes of transmission will necessitate proactive surveillance and predictive modelling as global temperatures rise and ecosystems change. By bridging the gaps between infectious disease dynamics and climatology, this research equips researchers to lessen future health emergencies. Interdisciplinary studies will develop exponentially over the next decade and the work makes sure they are based on solid, useful discoveries for a warming planet.

Acknowledgement

The authors are thankful to the Department of Biochemistry & industrial Microbiology, Sree Ayyappa College, Eramallikkara, Chengannur, Kerala and the Department of Botany, Nirmala College Muvattupuzha Autonomous, Ernakulam, Kerala for providing their facilities for this study.

Funding Sources

The author(s) received no financial support for the research, authorship, and/or publication of this article.

Conflict of Interest

The authors do not have any conflict of interest.

Data Availability Statement

This statement does not apply to this article.

Ethics Statement

This research did not involve human participants, animal subjects or any material that requires ethical approval.

Informed Consent Statement

This study did not involve human participants, and therefore, informed consent was not required.

Clinical Trial Registration

This research does not involve any clinical trials.

Author Contributions

Nitha Balan: Conceptualization and Writing Original Draft

Geena George: Analysis, Review & Editing

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Any product that is meant to be put, rubbed, sprinkled, sprayed, or applied on the human body or any part of it can be classified as cosmetic.¹ The use of synthetic products has been banned because of their carcinogenic properties by the Food and Drug supervisory committee.² According to ³, the most pervasive chemicals presented in cosmetics are in the form of preservatives, for example, paraben and sodium benzoate, also fragrances like limolene. Prolonged use of these chemicals has negative consequences on people’s health and well-being. ³ According to the USFDA’s tests in 2009 and 2011, several popular lipstick products contained lead (Pb) concentrations that exceeded the limit for safe cosmetics.⁴ (Campaign for Safe Cosmetics 2007) Synthetic UV filters used in sunscreens can cause bioaccumulation in several species and also they cause bleaching of coral, and hydrogen peroxide production in fishes thus harming marine life. This led Hawaii to prohibit certain sunscreens, including oxybenzone. Following suit, the Western Pacific nation of Palau, along with Key West, Florida, and the US Virgin Islands, also implemented similar bans.⁵ Additionally, heavy metals are sometimes used as coloring agents or preservatives in cosmetics; however, they are toxic and can negatively impact the nervous system, kidneys, and other organs. ¹ Due to the negative and harmful impacts of chemicals in cosmetics , and their environmental damages,⁴ there is a growing interest in natural and sustainable ingredients in cosmetics. People go in search of safe and natural ingredients to avoid side effects.⁶

Macroscopic algae, commonly known as seaweeds, play a crucial role in marine ecosystems. They typically thrive in shallow coastal waters and require a substratum to grow and develop. They lack true roots, leaves and stems. ⁷ Marine algae, categorized as red, brown, and green algae, have potential bioactive compounds with a wide range of applications, especially in skincare products.⁶.Brown, red, and green macroalgae contribute approximately 59%, 40%, and less than 1%, respectively, to the total global macroalgae cultivation.⁸

Recent discussions have highlighted the incorporation of algal substances in cosmeceuticals, focusing on their potential benefits for anti-aging, skin whitening, and skin cancer prevention. ⁹

Figure 1 : List of Seaweed Applications in Cosmetic Industry     Click here to view Figure

Bioactive compounds obtained from marine algae, including phenolic compounds, polysaccharides, pigments, PUFAs, sterols, proteins, peptides, and mycosporine-like amino acids (MAAs), possess diverse biological activities, making them valuable active ingredients in cosmetic formulations.⁶ This review comprises the use of seaweed-derived bioactive molecules in cosmetics and their novel formulation methods to increase their stability and efficacy.

Types of seaweed Bioactive compounds

Seaweeds generate a range of compounds through primary metabolism, known as primary metabolites, crucial for their growth, survival, and proliferation. These primary metabolites fall into various classes, including lipids, proteins, and carbohydrates. Beyond primary metabolites, algae also amass essential minerals vital for seaweed life, enhancing both its nutritional and pharmacological significance.¹⁰ In the realm of seaweed components, a diverse array of substances showcases remarkable properties in antiaging and antioxidant effects. Sulphated polysaccharides, peptides, carotenoids, fatty acids, and phytohormones stand out for their beneficial impacts on skin health. Additionally, mycosporine-like amino acids and flavonoids are noted for their antiphotoaging activity. Specifically, flavonoids, including phlorotannins, act as lipolytic agents derived from macroalgae, concurrently exhibiting inhibitory effects on melanogenesis, contributing to their multifaceted role in skin care.¹¹ Polysaccharides play a crucial role in various industrial applications, with sulfated types like fucoindans, carrageenans, and ulvan, as well as non-sulfated ones like alginates and agars, being the most widely recognized. Although certain polysaccharides like laminarin, xylans, porphyrans, argassan, and floridean are less abundant and not yet widely utilized in industry, ongoing research is exploring their potential applications for future use.¹⁰. Agar, a seaweed-derived ingredient, plays a vital role in numerous cosmetic products. Widely utilized as an emulsifier and stabilizer, agar ensures the consistent quality of creams. Its ability to control moisture content enhances the effectiveness of products such as hand lotions, deodorants, foundation, exfoliants, cleansers, and more. From shaving creams to anti-aging treatments, agar contributes to the stability and texture of formulations. Its versatility extends across various skincare and beauty items, highlighting its crucial role in the cosmetic industry.¹²

Table 1 : List of Seaweed and its Bioactive Compounds

Extraction methods and Processing techniques

There are different kinds of extraction methods depending upon the target compounds. Red algae extraction traditionally relies on energy-intensive HAE. ¹⁴ proposed HPAE as a greener, more efficient alternative. Employing pressure, HPAE achieves higher yields and lower extraction times compared to HAE, all at lower temperatures. This environmental advantage, coupled with its potential for green solvents, makes HPAE a compelling paradigm shift in red algae extraction, unlocking its bioactive potential sustainably. Advanced techniques such as Supercritical Fluid Extraction, Subcritical Water Extraction, Ultrasound-Assisted Extraction, and Microwave-Assisted Extraction are preferred over conventional methods due to their enhanced efficiency, selectivity, and sustainability.¹⁵ The conventional extraction method proved superior in phenolic and phlorotannin content, ultrasound-assisted extraction enhanced anti-elastase activity.¹⁶ Optimizing process parameters for each method is essential for obtaining extracts with the targeted bioactive compounds. ¹⁵

Antioxidant and  Radical Scavenging activity

The industries are moving towards natural antioxidants to replace synthetic antioxidants.¹⁷  Several authors analyzed the potential antioxidant activity of MAAs in vitro. In a study by Chen et.al.,¹⁸, an efficient degradation method for Sargassum fusiforme polysaccharides (PSF) using ascorbic acid and H2O2 was optimized. Under optimal conditions, degraded polysaccharides (DPSF) exhibited a DPPH radical scavenging rate of 75.22%. Notably, DPSF displayed superior antioxidant activities and tyrosinase inhibition compared to the original polysaccharide, indicating its potential for enhanced functionality.. It also has been proved that Ecklonia maxima exhibit promising potential in mitigating oxidative stress, diminishing melanogenesis, and thwarting photodamage in both invitro and invivo. ¹⁹

The study by Wang et al., found that SF-F4 was effective in increasing the viability of PM-treated HaCaT cells by inhibiting apoptosis and scavenging intracellular ROS.²⁰ Additionally, In another study, PBP (Padina boryana ethanol precipitate) extracted from the marine brown alga Padina boryana was evaluated as a potent natural antioxidant. The research delved into the chemical composition of PBP, emphasizing the contributions of sulfate content, fucose, and galactose to its bioactive properties. The study highlighted PBP’s exceptional potential in safeguarding against ROS-induced cell damage and mitigating oxidative stress in zebrafish. Notably, the upregulation of Nrf2 and subsequent elevation of CAT and SOD protein levels shed light on the underlying mechanisms of PBP’s antioxidative effect.²¹ A study by Kang et al. ²² examined the antioxidant and whitening activities of a fermented extract comprising Undaria pinnatifida, Saccharina japonica, and Gloiopeltis furcate. The physiological effects of combined seaweed extracts were analyzed using Lactobacillus sakei strains derived from kimchi as lactic acid bacteria. Notably, the antioxidant potential, assessed through DPPH and ABTS radical scavenging assays, demonstrated that the inhibitory effects of the combined seaweed extracts surpassed those of the positive control, indicating enhanced antioxidant activity.²² Thus seaweed extracts can be used as an anti-oxidant in cosmetics.

Collagenase inhibition and Anti-Aging benefits

Seaweed extracts present a compelling avenue in the pursuit of natural anti-aging strategies. Polysaccharides, particularly fucoidans and their sulfated brethren, exhibit remarkable activity in dampening collagenase and elastase activity, thereby protecting the intricate tapestry of the skin matrix and mitigating wrinkle formation ⁶. This protective shield extends beyond polysaccharides, as recent investigations by ¹⁶ revealed a treasure trove of novel anti-aging metabolites within the enigmatic Sargassum horridum. Notably, Diosgenin, a steroidal saponin, alongside a quartet of previously unreported phenolic compounds, collectively address skin elasticity loss and photoaging triggered by reactive oxygen species, paving the way for the development of cutting-edge, seaweed-based anti-aging formulations.

Skin whitening

Many people in Asia desire a fair and flawless complexion and often seek skin whitening treatments to achieve it. In the process of melanin synthesis, tyrosinase is a crucial enzyme that plays two significant reactions. First, it hydroxylates l-tyrosine to 3,4-dihydroxy-l-phenylalanine, which further gets oxidized to dopaquinone. Then, dopaquinone is further converted to melanin. Sun exposure can increase the synthesis of both tyrosinase and melanosomes, which can lead to an increase in melanin production, resulting in darker skin.²³ A study demonstrated that Sulfated polysaccharides from Celluclast-assisted extract of Hizikia fusiforme (HFPS) displayed anti-melanogenic effects by down-regulating tyrosinase and TRP-1 and -2, thereby inhibiting melanin synthesis. These findings suggest the potential of HFPS in both pharmaceutical and cosmeceutical industries for skin whitening.²⁴ . An in vitro study evaluating tyrosinase inhibition demonstrated that S. siliquosum exhibits superior inhibitory activity, boasting an IC50 value of 65.0 μg GAE ml−1. This surpasses the efficacy of the well-known skin-lightening ingredient kojic acid, which recorded an IC50 value of 109.32 μg GAE ml−1. These findings highlight the potential of S. siliquosum as a promising candidate for skin-lightening applications, potentially outperforming established compounds in terms of tyrosinase inhibition.²⁵ Another study aimed to investigate the anti-melanogenesis effect of Sargassum polycystum extracts by conducting various assays using B16F10 murine melanoma cells. SPHF inhibited melanogenesis by inhibiting cellular tyrosinase activity and may be useful for treating hyperpigmentation.²⁶

Hydration and Moisturization

Moisturizer agents help to maintain skin appearance and elasticity, improving its barrier role against harmful environmental factor.²⁷ Seaweeds, known for their rich amino acid content, including arginine, and a plethora of vitamins (A, B, C, D, and E), offer notable moisturizing benefits for the skin, contributing to its elasticity and acting as a barrier against environmental factors.²⁸ Polysaccharides like alginate, agar, carrageenan, and fucoidans derived from specific algal species play a crucial role in regulating water distribution in the skin,²⁸ with studies demonstrating impressive moisturizing rates, particularly in marine green algae such as Enteromorpha prolifera and Enteromorpha linza.²⁹ Fucose, present in glycoproteins, not only provides moisturization but also exhibits anti-aging effects ³⁰. Seaweed polysaccharides, such as those extracted from Saccharina japonica, have been found to surpass hyaluronic acid in moisturizing properties, suggesting their potential as valuable cosmetic ingredients ³¹. The lipid composition of seaweeds, including polyunsaturated fatty acids (PUFA) like γ-linolenic acid, arachidonic acid, eicosapentaenoic acid, and docosahexaenoic acid, along with sterols and phospholipids, enhances the skin barrier, offering protection.³² Agarobiose , a seaweed-derived component, serves as a moisturizer for both skin and hair, highlighting the multifaceted benefits of seaweed-based ingredients in skincare formulations. ³³

Acne aggravation, eczema, physical texture arise if the skin is not properly moisturized²⁸ Seaweed fulvescens (SF) is a green alga rich in chlorophyll . Treatment at 200 mg/mouse demonstrated inhibitory effects on AD ( Atopic dermatitis) symptoms, leading to improved dorsal skin conditions, reduced inflammation, smaller lymph nodes, and lower levels of proinflammatory cytokines. In HaCaT keratinocytes, SF (10, 25, and 50 μg/mL) dose-dependently suppressed the production of proinflammatory cytokines and reduced the phosphorylation of signal transducer and activator of transcription . These findings highlight the potential therapeutic benefits of SF in the treatment of Atopic deramatitis positioning it as a promising alternative.³⁴ In a groundbreaking study investigating the modulation of reactive skin microbiota, the positive impact of Halymenia durvillei (HD), an active ingredient rich in polysaccharides, was evident over a concise 28-day period in 30 volunteers with reactive, sensitive skin. Notably, the analysis of skin microbiota showcased a predominance of beneficial Actinobacteria, Proteobacteria, Firmicutes, and Bacteroidetes, with no adverse changes observed post-HD treatment. The study underscored the remarkable maintenance of microbial communities in reactive skin, emphasizing the potential benefits of HD for promoting skin health. Additionally, clinical improvements aligned with a decrease in Corynebacterium kroppenstedtii, a marker associated with redness. Furthermore, an ex vivo assessment revealed the rapid and significant reduction in neuroinflammation parameters after just 6 days of HD extract application. These encouraging results highlight the promising role of HD polysaccharides in managing and enhancing the well-being of reactive and sensitive skin, shedding light on their positive impact on skin microbiota and neuroinflammation.³⁵

Anti-Microbial Effects

The antimicrobial properties of seaweeds have been known since ancient times and well-documented in recent years and thus they are of interest for potential use in cosmetic products. ³⁶. In a study conducted by  ³⁷   F2 and F7 fractions of Fucus spiralis exhibited the highest inhibition against C. acnes and M. furfur, respectively, leading to approximately 50–60% reduction in the growth of these microorganisms. Incorporating Sargassum polycystum ethanol extract as a natural preservative in sunscreen offers a compelling alternative to synthetic counterparts. Comparative studies reveal its antimicrobial efficacy, matching methylparaben’s 8-week preservation. The formulated cream demonstrates stability up to one year, balanced pH, absence of foul odor, an IC50 antioxidant activity of 105.42, and an SPF value of 2.00. These results highlight the potential of S. polycystum in cosmetics, aligning with the demand for effective, natural, and sustainable skincare solutions.³⁸

Sargassum vulgare alginate, extracted post-ethanol pretreatment, showed superior antimicrobial efficacy compared to herbal preservative 705. Notably effective against Pseudomonas aeruginosa and Staphylococcus aureus within a shorter timeframe, suggesting its potential for cosmetics applications.³⁹

UV Protection and Photoprotection

Mycosporine like aminoacids are compounds that has UV-absorbing properties to protect from UV induced damage. MAAs are small, water-soluble compounds with a molecular weight typically below 400 Da. These colorless molecules exhibit high stability under various environmental conditions. Structurally, they consist of either an aminocyclohexenone or aminocyclohexenimine ring, featuring nitrogen-containing substituents.⁵ MAA are water soluble and their absorption maxima is between 265nm and 362nm. These molecules are excellent UV-absorbing compounds with low toxicity, especially high stability and good antioxidant activity.⁴⁰ Their concentration fluctuates throughout the year due to the effect of insolation.

Conventional sunscreens which use organic or inorganic UV filters has the capability to produce ROS on the skin .But MAA when used as UV filters in sunscreen has antioxidant properties which can retaliate the ros accumulation caused by the conventional UV filters. Porphyra-334 isolated from Porphyra yezoensis showed a protective effect on human skin fibroblasts exposed to UV-A radiation, increasing cell viability up to 88%. It also inhibited the accumulation of ROS in human skin fibroblasts damaged by UVA-induced oxidant stress in a dose-dependent manner, with similar results seen at 40 µM. ⁴⁰ PBP demonstrated significant photoprotective effects in an anti-photodamage test, showcasing its ability to protect skin cells from UVB-induced damage. Specifically, PBP inhibited apoptosis and reduced intracellular reactive oxygen species levels in human epidermal keratinocytes (HaCaT cells) following UVB irradiation. The reduction in reactive oxygen species levels was dose-dependent, with 25 μg/mL, 50 μg/mL, and 100 μg/mL of PBP showing varying degrees of efficacy. Additionally, PBP exhibited notable protective actions on human dermal fibroblasts, including the suppression of oxidative damage, inhibition of collagen degradation, and attenuation of inflammatory responses. These findings highlight PBP as a promising candidate for photoprotective applications in skincare formulations.¹⁹

Anti-inflammatory Effects on skin

A recent study by Wang et al., ²⁰  aimed to investigate the potential of SF-F4, a fucoidan extracted from S. fusiforme, to prevent skin damage caused by Particulate matter (PM)  exposure. The study revealed that SF-F4 effectively enhanced the viability of PM-treated HaCaT cells by preventing apoptosis and reducing intracellular ROS levels. Furthermore, SF-F4 regulated the expression of MMPs and pro-inflammatory molecules in PM-stimulated HDF cells, leading to an increase in pro-collagen content. These findings suggest that SF-F4 has significant potential in preventing skin damage caused by PM exposure and can be a valuable ingredient in pharmaceutical and cosmetic products. ²⁰ In another study by Shih et.al ⁴¹, researchers explored the potential of low-molecular-weight fucoidan (LMF) as a supplement for treating atopic dermatitis . (AD) LMF  was prepared from Sargassum hemiphyllum which is known for its anti-inflammatory properties. The results revealed that the group supplemented with LMF experienced significant relief in AD symptoms. Notably, the frequency of using steroid ointments and oral antihistamines decreased in the LMF group, suggesting reduced inflammation.⁴¹ Additionally, Wang et al. ²⁷ investigated the antioxidant and anti-wrinkle effects of sulfated polysaccharides from Celluclast-assisted extract of Hizikia fusiforme . (HFPS) Their findings suggested that HFPS could be a promising candidate for cosmeceutical applications due to its significant anti-inflammatory effects, including the inhibition of nitric oxide generation, reduction of pro-inflammatory cytokines, and suppression of iNOS and COX-2 expression in stimulated macrophages.²⁷

Formulation Strategies in Cosmetic Science

Seaweed components have been effectively incorporated into various physical forms and are commercially available in products such as soaps, shampoos, sprays, hydrogels, and creams.⁴² Their effectiveness and stability can be enhanced using appropriate carrier systems or vesicles, such as liposomes, nano/microparticles, emulsions, and hydrogels, which are designed to deliver active agents in commercial formulations for improved performance.⁴³

Nanosystems have been used to encapsulate the seaweed bioactive molecules thereby  enhancing the  stability and efficacy of  the cosmetic  formualtions . Nanosystems  can help in sustained release of the product , permanence on the skin for a long time , minimizing the active ingredient hence avoiding the toxic effects caused by higher concentrations.⁴³ . Hu et al.⁴⁴ developed a straightforward approach to fabricate hydrophilic-hydrophobic core-shell microparticles utilizing seaweed-derived polymers. These microparticles hold promise for applications in safeguarding unstable compounds and enabling the controlled release of drugs or bioactive ingredients in cosmetic formulations Seaweed polymers provide significant advantages in terms of biocompatibility and biodegradability.³⁹

Challenges and Future Directions

Consumers often lack awareness that natural-based cosmetic products comprise a intricate blend of both natural raw materials and chemical compounds, potentially leading to adverse effects on human health ⁴⁵ . Utilizing algae as a cosmeceutical ingredient presents certain challenges, including concerns related to (i) biomass culturing techniques, (ii) metabolite extraction methods, and (iii) ensuring quality assurance and compliance with regulations . ⁴⁵ Seaweeds have a tendency to accumulate heavy metals like cadmium (Cd), coper (Cu), manganese (Mn), niquel (Ni), lead (Pb), zinc (Zn), mercury (Hg) from water. So, the applications of seaweed in cosmetics  should be accompanied by chemical  analysis to evaluate the  safety of the raw materials. ³⁰

Conclusion

An interest on natural products is increasing in cosmetic industry. In that sense, seaweed is a natural source of many bioactive molecules like fucoidans , alginates , careeganan , MAA which are a great replacement for conventional or chemical active ingredients used in the formulations. Along  with functional bioactives ,seaweed lipids and polysaccharides   can also be used as nanocarriers for the bioactive molecules which can enhance the stability and efficacy of the product. Future prospects to look for are enhancing the production of certain bioactive molecules , optimizing the extraction process to increase the yield , utilizing the stabilizing capacity of the seaweed lipids and using it as carriers , more clinical studies need  to be carried out to determine and assure the safety and improve the quality of the product.

Acknowledgement

The author would like to thank Annamalai University for granting the Ph.D. research work.

Funding Sources

This research was funded by the RUSA 2.0 Research & Innovation – Health and Environment Scheme under the project entitled ‘To study the climate on antibiotic resistant pattern of bacterial pathogens’, approved under Field 5: Marine Ecosystem Assessment . (Order No. DRD/RUSA 2.0/R&I/Project Proposal/Field 5/2021, dated 31.01.2022) We acknowledge the support of the Rashtriya Uchchatar Shiksha Abhiyan (RUSA) and the Higher Education Department.

Conflict of Interest

The authors do not have any conflict of interest.

Data Availability Statement

This statement does not apply to this article.

Ethics Statement

This research did not involve human participants, animal subjects, or any material that requires ethical approval.

Informed Consent Statement

This study did not involve human participants, and therefore, informed consent was not required. 

Clinical Trial Registration

This research does not involve any clinical trials.

Permission to reproduce material from other sources

Not Applicable 

Author Contributions

Gayathri Sivakumar: Conceptualization, Literature Review, Writing – Original Draft, Visualization.

Sivasubramani Kandasamy: Supervision, Writing – Review & Editing, Project Administration, Resources.

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Seizures represent a sudden and transient neurological event characterized by abnormal, excessive electrical activity within a population of central nervous system neurons. This rapid, synchronous firing can arise from various etiologies, including epileptic conditions, systemic metabolic disturbances (e.g., hypoglycaemia, hypercalcemia), intracranial infections, severe systemic infections, or as an adverse effect of certain medications. Epilepsy encompasses a spectrum of chronic neurological disorders defined by the recurrent occurrence of seizures, often accompanied by alterations in consciousness and/or characteristic motor manifestations (convulsions). Autonomic dysfunction may also be observed in some cases.¹

Epilepsy is a prevalent and significant neurological disorder, affecting approximately 1% of the global population. A considerable subset of these individuals, around one-third, experience refractory epilepsy, characterized by seizures that are not adequately managed with standard antiepileptic drugs. Notably, the onset of epilepsy occurs in about 75% of cases during childhood, underscoring the increased vulnerability of the developing brain to epileptic events. However, in developed nations, the incidence of childhood epilepsy has declined over the past thirty years, while there has been a corresponding rise in the prevalence among the elderly population.^2-5

Epilepsy is a neurological disorder impacting individuals across different age groups, characterized by abnormal electrical activity in parts or throughout the brain. This condition manifests as seizures, which can occur spontaneously and recurrently. Seizures may result from a variety of factors, including stroke, brain tumours, head trauma, or central nervous system infections. It is estimated that approximately fifty million people worldwide are currently living with epilepsy, and this disorder accounts for about one percent of the global disease burden, with higher prevalence in low- and middle-income countries. Despite the availability of numerous pharmacological treatments for epilepsy, a significant number of patients remain unresponsive or develop resistance to antiepileptic drugs, making pharmacoresistance a major clinical challenge in the treatment of epilepsy.^6-9

Epilepsy is a prevalent, severe, chronic, and potentially life-shortening neurological disorder marked by a continuous predisposition to seizures. It impacts over 60 million people worldwide and is a significant contributor to seizure-related mortality, comorbidities, disabilities, and healthcare costs. Various treatment strategies have been employed to manage epilepsy, and over 30 drugs have received approval from the US FDA. However, approximately 25% of individuals with epilepsy remain resistant to these treatments. Approximately 90% of people in low- and middle-income countries lack access to modern epilepsy medications. In these regions, traditional remedies such as plant extracts have been employed to treat various conditions, including epilepsy. These medicinal plants offer significant therapeutic potential due to their valuable phytochemical content with a range of biomedical applications. Given that epilepsy is a multifactorial disorder, multitarget approaches using plant extracts or isolated phytochemicals are necessary, as they can address multiple pathways simultaneously. Various plant extracts and phytochemicals have demonstrated efficacy in treating epilepsy in different animal models by interacting with diverse receptors, enzymes, and metabolic pathways. These natural compounds hold potential for future use in human epilepsy treatment. However, more research is required to elucidate their precise mechanisms of action, assess toxicity, and determine appropriate dosages to minimize side effects.^10-17

Types of epilepsy.^{1, 18-24}

Primarily generalized seizures

Generalized tonic-clonic seizures (Grand mal)

Tonic seizures

Atonic seizures

Absence seizures (Petit mal)

Myoclonic seizures

Partial seizures (Focal or local seizures)

Simple partial seizures

Complex partial seizures

Secondary generalized seizures

Primarily generalized seizures

Generalized tonic-clonic seizures (Grand mal epilepsy)

These are also called as grand mal epilepsy / major epilepsy. One of the serious type of convulsions which consist of maximum tonic spasms of all body muscles then instantaneous jerking moments followed by central depression. Tonic-clonic seizures cause instant loss in awareness (consciousness) without any signs then tonic and then clonic convulsions. Drowsiness, sleep and headache are the minor symptoms of the attack. Convulsive moments also include frothing, tongue biting and urinary incontinence. When the term “epileptic fit” is used informally, it usually refers to this kind of seizure.

Tonic seizures

Tonic seizures, also known as opisthotonus, cause significant autonomic symptoms and unconsciousness. Without the clonic phase, it resembles tonic-clonic seizures.

Atonic seizures

Head slumping, falling, and/or loss of postural tone are the hallmarks of atonic seizures. Loss of muscular tone during atonic seizures results in the person falling to the ground. These are commonly referred to as “drop attacks,” although they should be differentiated from similar-looking events that can happen in people with cataplexy or narcolepsy.

Absence seizures (Petit mal)

An interruption of consciousness known as an absence seizure, occurs when the person having the seizure appears vacant and unresponsive for a brief moment (often up to 30 seconds). There may be a slight twitching of muscles.

Typical absence seizures

Atypical absence seizures

Myoclonic seizures

These are clonic jerks associated with short bursting of many spikes in electroencephalogram. Muscles or muscle groups may jerk during myoclonic seizures, which are characterized by a very short (<0.1 second) contraction of the muscles.

Partial seizures

These are also called as focal seizures / local seizures. There are two types of partial seizures i.e. simple partial seizures and complex partial seizures.

Simple partial seizures

There are several ways that simple partial seizures might appear without impairing consciousness. Depending on the precise cortical area generating the aberrant discharge, they may include unique and localized sensory problems (Jacksonian sensory epilepsy), convulsions restricted to a single limb or muscle group (Jacksonian motor epilepsy), and other limited signs and symptoms. Simple seizures may result in sensory abnormalities or other symptoms, but they do not disrupt consciousness.

Complex partial seizures

These attacks cause disorientation and awareness impairment. In addition to a variety of clinical symptoms, such as odd worldwide EEG activity during the seizure, they frequently show signs of localized abnormalities in the front temporal lobe during the inter-seizure phase. Complex seizures cause varied degrees of awareness disruption. This does not imply that the individual having this kind of seizure will faint or become unconscious.

Secondary generalized seizures

These seizures begin as simple partial seizures, which progress to become generalized seizures. Todd’s paralysis, a postictal neurological impairment, may occur after such a seizure. Jacksonian seizures occur when the motor phenomena spread in an orderly fashion.

Status epilepticus (SE)

SE is characterized by repeated seizures of any kind or prolonged seizures (lasting more than five to ten minutes) with no recovery in between bouts. When tonic-clonic seizures develop into the SE, which might be lethal, a medical emergency arises. The aforementioned explanation pertains to a category called “seizures.” However, the classification of “epilepsies” also has to consider the prognosis, responsiveness to treatment, age of onset, hereditary variables, EEG abnormalities, concomitant neurologic deficits, and imaging results. When choosing a medication and assessing prognosis, it may be more beneficial to define a particular epilepsy syndrome rather than focusing solely on seizure characteristics. Approximately 10% of people with actual seizures have several EEG examinations that show no abnormalities. Therefore, in a person with a definitive clinical picture, there is no way to rule out a seizure illness with a normal EEG.

Classification of anti-epileptic drugs (AEDs).^18,25-27

Classification according to chemical Nature

Barbiturate: Phenobarbitone.

Hydantoins: Phenytoin, Mephenytoin, Phenyl ethyl hydantoin.

Oxazolidinediones: Trimethadone, Paramethadione.

Phenacemide: Phenacemide, Phenyl ethyl acetyl urea.

Benzodiazepines: Nitrazepam, Clonazepam.

Iminostilbenes: Carbamazepine.

Miscellaneous: Sodium Valproate (Valproic acid)

Classification according to Mode of Action:

Modulation of Ion Channels: Phenytoin, Carbamazepine, Lamotrigine, Oxcarbazepine, Ethosuximide, Zonisamide.

Potentiation of γ-amino Butyric Acid: Phenobarbital, Benzodiazepines, Vigabatrin, Tiagabine.

Drugs with multiple mechanism of action: Sodium Valproate, Gabapentin, Felbamate, Topiramate.

Drugs with unknown mechanism of action: Levetiracetam. 

Pharmacological targets of AEDs.^18,28-35

Figure 1: Pharmacological targets of the AEDsClick here to view Figure

Voltage dependent ion channels are targets for many antiepileptic drugs. These are Na⁺, Ca⁺ and K⁺.

Ion Channels

Na+ channels:

These ion channels regulate movement of cations across internal to external cell membranes. The multi-subunit structure of neuronal sodium channel creates voltage-gated, Na+-selective pore across the plasma membrane. Conductance via the intrinsic pore is controlled by conformational changes in the protein structure in response of variations in the membranous potential. Primary structural element of the sodium channels of nervous system is subunit-A. Another two subunits like subunit-β1 and subunit-β2, form associations with α-subunit in the mammalian brain. Subunit- β1 and subunit-β2 are not necessary for the sodium channel activity. The majority of Na+ channels are in a closed, resting state at typical membrane potentials. Ion flux is facilitated by the channel’s activation upon depolarization. The Na+ channel then goes into an inactivated state from which it is difficult to restart. The channel quickly returns to a permanent state upon repolarization of the neuronal membrane, from which it can react to further depolarizations.

Ca2+

These are analogue of voltage gated sodium channels. The analogue of α1-subunit of sodium channel is subunit- α of Ca2+ channel. This adds voltage dependency and creates the Ca2 + sensitive channel pore. Ca2+ channels are classified as either low threshold or high threshold based on their membrane potential. It is believed that the minimal-threshold T-type calcium channel, which mostly found in relay neurons in the thalamocortical region, is responsible for the rhythmic 3-Hz spike-and-wave discharge that is characteristic of generalized absence seizures. Based on their pharmacological characteristics, high-threshold Ca2+ channels divided in type P-, R-, Q-, N-, and type L-. All channels are found on cell bodies, nerve terminals and dendrites throughout CNS. Regulation of neurotransmitter release at synapse has been linked to the P-, Q- and N-type. In order to contribute to anti-epileptic medications, many AEDs work by inhibiting voltage-sensitive Ca2+ channels.

K + channels

In the large complexes of protein, K+ channels are in the form of tetramer, and their monomers share structural and genetic similarities with the Ca+ and Na+ channels via the α1 and α1 subunits, respectively. The excitation involves K+ channels. They are in charge of the Na+ channel’s plasma membrane repolarization. Direct voltage-dependent K+ channel activation inhibits action potential firing and hyperpolarizes the neuronal membrane. K+ channel blockers cause seizures, while potassium channel facilitator shows antiepileptic benefits in several experimental animal models. AED development in the future may focus on acceleration of potassium ion channel currents.

γ- Aminobutyric acid-mediated inhibition

GABA, central nervous system’s main inhibitory neurotransmitter.

It is well known that seizures are triggered by impairment of GABA function, while stimulation has an anticonvulsant effect. The enzyme glutamic acid decarboxylase is responsible for the synthesis of GABA in GABAergic neurons. GABA is crucial for regulating both the excitatory output from the cortex and glutamate-mediated excitatory activity within it. GABA receptors come in two subtypes, known as GABAA and GABAB, as well as the recently identified GABAC. Fast neurotransmission is facilitated by GABAA receptors, which are mostly found on postsynaptic membranes. The ligand-gated ion channel superfamily includes it and causes neuronal hyperpolarization in response of binding of GABA by conductance of chloride ions. K+ conductance rises when G-protein coupled receptors like GABAB gets activated. The transmitter gated channel, which forms the pore from which Cl^– ions comes in the neuron of post synapse, after GABAA receptor occupation, is made up of five membrane-spanning subunits, including the GABAA receptor. Four different transmembrane spanning domains make up each of the five subunits. α, β, γ, δ, and ρ are the thesis subunits that make up the ionophore; each of these subunits, except for δ, has several isomeric forms; 3 y subunits (γ1-γ3), 4 β subunits (β1~β4), and 6 subunits (αl-α6) are present. one subunit of δ, and 2 subunits of ρ i.e.  (ρl-ρ2), It appears to be situated in the retina. GABA’s synaptic activity is stopped by GABA transporters found on glial cells and presynaptic nerve terminals. There are about 4 known GABA transporter proteins. These proteins include GAT-1, GAT-2, GAT-3 and BGT-1. Sodium and chloride gradients of transmembrane are necessary for the functions of GABA transporters.

Glutamate (GLUT) Mediated Receptor

The main excitatory neurotransmitter in the neurological system is glutamate. Animal experiences epileptic seizures when they receive glutamate’s focal injections.    Several epilepsy disorders and animal seizure models exhibit aberrant glut receptor characteristics or over-activation of glutamatergic transmission. Numerous receptors are affected by glutamate’s pharmacological actions. Glutamate is changed into glutamine by glial cells using the enzyme glutamine synthetase. making glial glutamate uptake crucial. The cycle is then completed when glutamine is delivered to glutamatergic neurons. Ionotropic receptors of glutamate, like GABA receptors, made up of the several subunit combinations which forms arrays of tetrameric and pentameric forms. Three separate subtypes are distinguished among them: N-methyl-D-aspartate (NMDA), α-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid (AMPA) and kainite. These subtypes produce ion channels that are ligand-gated that permit the passage of sodium also contain Ca2+ ions based on the subtype and subunit composition. Glycine’s role as a co-agonist further distinguishes NMDA receptor. Glutamate receptor subtypes AMPA and kainate are linked to rapid excitatory transmission of neurons. While another subtype i.e. NMDA which is dormant at the resting membrane potential which activated by extended depolarization.

Screening models of epilepsy.^36,37

In vivo methods

Maximal electroshock induced convulsions in mice.^38-40

The main aim of the electroshock method in animals used is to identify substances that are useful in treating grand mal epilepsy. Anti-epileptic medications and other centrally active medications block the electric impulses that cause tonic hindlimb extensions.

Procedure

NMRI mice (Male) weighing 18–30 gram are utilized in groups of 6–10. The test begins thirty minutes after i.p. injection / sixty minutes Upon oral administration of vehicle or test chemical. The stimuli are delivered using a device that has electrodes in the cornea or ears (Woodbury and Davenport 1952). The device determines the stimulus’s intensity; for example, 12 mA and 50 Hz for 0.2 s have been employed. All mice given the vehicle exhibit the distinctive extensor tonus under these circumstances.

Evaluation

For two minutes, the animals are closely watched. The lack of the hindleg extensor tonic convulsion is the condition for affirmation. The percentage of seizures that are inhibited in comparison to controls is computed. Probit analysis is used to get ED50 values and the 95% Confidence interval using different doses.

Pentylenetetrazol (Metrazol) induced convulsions.^39-44

The main aim of this test is to assess anti-epileptic medications. Anxiolytic medications can likewise to stop or counteract convulsions brought on by Metrazol.

Procedure

Both sexes of mice having weight between 18 and 22 grams are employed. Groups of ten mice receive oral, intraperitoneal, or sc. injections of the test substance or reference medication. A control group of ten mice is used. 30 minutes following intraperitoneal injection, 15 minutes following sc. injection, or 60 minutes following oral delivery Metrazol (60 mg/kg) is administered subcutaneously. Every animal is kept in a separate plastic cage for a one-hour observation period. Tonic-clonic convulsions and seizures are documented. The control group’s animals must exhibit convulsions in at least 80% of cases.

Evaluation

The percentage of afflicted animals in the control group is used to determine the quantity of animals in the treated groups that are protected. It is possible to compute ED50 values. Additionally, it is possible to measure the interval between the metrazol injection and the onset of seizures. The onset delay is computed compared to the control group.

Strychnine-induced convulsions.^45,46

Strychnine causes convulsions via interfering with glycine-mediated postsynaptic inhibition. Strychnine works as a competitive, selective antagonist to prevent glycine from inhibiting any glycine receptor. Glycine is crucial inhibitory transmitter to interneurons and moto-neurons in spinal cord. Glycine also mediates postsynaptic inhibition which is strychnine-sensitive in the higher centres of central nervous system. It has been demonstrated that compounds that counteract strychnine’s effects have anxiolytic effects.

Procedure

10 mice of each sex with wt. about 18 to 22 grams, which utilized in groups. Test substance / std one (such as 5 mg per kg of diazepam) is administered orally to them. After an hour, the mice get an i.p.  injection of strychnine nitrate at dose 2 mg per kg. A period of 11 is used to record the duration till death and tonic extensor convulsions. Eighty percent of the controls experience convulsions when given this dosage of strychnine.

Evaluation

Using several doses and a 100% control percentage, ED50 values are computed. The duration between treatment and strychnine injection for time response curves ranges from 30 to 120 minutes.

Isoniazid-induced convulsions.^47,48

Patients with seizure disorders may experience convulsions as a result of taking isoniazid. The substance is thought to be an inhibitor of GABA production. Mice experience clonic-tonic convulsions, which anxiolytic medications counteract.

Procedure

The test substance or std one (e.g., diazepam 10 mg per kg intraperitoneally) will administer orally or i.p. to ten either sex mice having weight between 18-22 grams. The controls only receive the vehicle. Mice get a s.c. injection of 300 mg per kg INH thirty min after intraperitoneal or sixty min after postoperative therapy. Death, tonic seizures, and clonic seizures are noted throughout the course of the following 120 minutes.

Evaluation

The control group is assumed to have a 100% chance of seizures or death. The percentage of controls is used to determine how much these effects are suppressed in the treated groups. Values for ED50 are computed.

Picrotoxin-induced convulsions.⁴⁹

CNS-active substances are further assessed by inducing convulsions with picrotoxin. It is believed that picrotoxin is a GABA-antagonist that alters GABA receptor complex’s Cl^– ion channel activity.

Procedure

Test substance or std one (example ten mg per kg diazepam intraperitoneal) is administered orally or intraperitoneally (i.p.) to 10 mice of either sex having wt. between 18-22 grams. Animals get an injection of 3.5 mg per kg subcutaneous picrotoxin 30 minutes after intraperitoneal treatment or sixty minutes after oral delivery. Over the next 30 minutes, they are monitored for the symptoms listed: tonic seizures, clonic seizures and death. Seizures starting time and death time are noted.

Evaluation

The animals are given the medication 30, 60, or 120 minutes before picrotoxin in order to obtain time-response curves. % inhibition in relation to the vehicle control is how protection is expressed. The peak time of pharmacological activity is defined as the time period having the highest percent inhibition. When calculating ED values, the control group’s seizure percentage is set at 100%.

Bicuculline test in rats.⁵⁰

Bicuculline, a GAGA-antagonist, can cause seizures, while well-known anti-epileptic medications counteract them.

Procedure

Bicuculline at a dose of one milligram per kilogram is administered intravenous to Sprague-Dawley rats (females). Within 30 seconds of injection, all treated rats experience a tonic convulsion at this dosage. The test chemicals are taken orally one or two hours prior to the injection of bicuculline. It is possible to get dose-response curves.

Evaluation

The % of animals that are secured is assessed. Probit analysis is used to determine ED-values and 95% confidence limits.

4-aminopyridine-induced seizures in mice.⁵¹

4-aminopyridine, antagonist of K⁺ channel, causing convulsions in both humans and animals. The medication passes through the blood-brain barrier with ease and is thought to increase both evoked and spontaneous neurotransmitter release, which in turn causes seizure activity. 4-aminopyridine facilitates both inhibitory and excitatory transmission at synapse; nevertheless, non-NMDA type excitatory amino acid receptors are primarily responsible for the drug’s epileptiform action. When 4-aminopyridine is given parenterally to mice, it causes clonic-tonic seizures and death.

Procedure

NIH Swiss mice (Male) with weight 25 to 30 grams are let to become accustomed to open access to water and food for a twenty-four hrs duration pre-testing. 15 minutes before the intraperitoneal injection of 4-aminopyridine at dose of 13.3 milligram per kilogram. Test medicines are given in different doses. Only typical behavioural symptoms, including hyperreactivity, shaking, intermittent hindlimb/forelimb clonus then tonic seizures, opisthotonus, hindlimb extension and death are seen in controls treated with 4-aminopyridine. At the LD97, the average latency to death is roughly ten minutes. Each dose is given to eight mice group.

Evaluation

ED50 values are determined using the proportion of safeguarded animals for every dosage. Broad-spectrum anticonvulsants like phenobarbital and valproate, as well as phenytoin-related anticonvulsants like carbamazepine, are helpful. In contrast, GABA-enhancers like diazepam, CA2+ channel antagonists like nimodipine and a few of NMDA antagonists are not.

Epilepsy induced by focal lesions.⁵²

epilepsy may be caused in animals by intrahippocampal injections of toxic substances or certain brain injuries. Rats given intrahippocampal kainic acid injections had their long-term effects examined by Cavalheiro (1982).

Procedure

A stereotactic setup is used to anesthetize adult Wistar rats (Male) using a chloral hydrate/nembutal mixture. A burr hole into the calvarium is used to implant a 0.3 mm cannula for injections. Hippocampal injection coordinates are derived from a stereo-tactic map, such as Albe-Fessard (1971). In a volume of 0.2 µl, kainic acid is dissolved in the fake serum and administered into different doses (0.1-3.0 µg) over a 3-minute period. For recording, 100 µm bipolar twisted electrodes are stereotaxically placed and cemented to the skull with dental acrylic cement. Two sites for depth recording are the dorsal hippocampus and the amygdala ipsilateral to the injected side. Jeweler’s screws are used to orient surface electrodes across the occipital brain. For grounding, an additional screw serves as an indifferent electrode in the frontal sinus.  EEG polygraph records the signals.

Evaluation

Both with and without pharmacological treatment, while the acute phase and the chronic phase (up to two months).  EEG recordings and convulsive seizure observations and made.

Genetic animal models of epilepsy.³⁶

Dogs, rats, and mice are among the animal species that exhibit convulsions and spontaneous repeated jerky moments. Double-mutant spontaneous epileptic rats that have both absence-like and tonic seizures were described by Serikawa and Yamada (1986).

Procedure

The tremor heterozygous rat (m/+) and the zitter homozygous rat (zi/zi) from a Sprague-Dawley colony are mated to produce spontaneous epileptic rats. Every week, two hours of the spontaneous epileptic rats’ behaviour are captured on videotapes. When no outside stimuli are present, the incidence of tonic seizures and frantic hopping is seen. The hippocampus and left frontal cortex are permanently implanted with monopolar stainless-steel electrodes and silver ball-tipped electrodes under anaesthesia. The frontal skull is exposed to an indifferent electrode. The EEG measures the duration of each seizure and the frequency of tonic convulsions and absence-like seizures. To produce regular tonic convulsions, a gentle tactile stimulus is applied to the animal’s back every 2.5 minutes. I.P. or oral administration of compounds.

Evaluation

Every five minutes before and after the drug injection, the number of seizures and their durations are recorded, and the sum of the convulsions’ durations (number into duration) computed. Values both prior to and following drug administration are compared as a percentage.

Kindled rat seizure model.⁵³

Goddard (1969) were the first to describe kindling, which is caused by repeated electrical stimulation of specific brain areas that causes sub-convulsions. Local after-discharge is initially linked to modest behavioural symptoms; however, generalized convulsions are likely to arise as electrical activity spreads with sustained stimulation. Despite the incomplete understanding of the pathophysiology of kindling seizures, it is a valuable way to examining effectiveness of antiepileptic medications.

Procedure

The Sprague-Dawley rats utilized are adult females weighing between 270 and 400 g. Pellegrino (1979) used the following coordinates to implant an electrode into the right side of rats’ amygdala: horizontal, 2.5, lateral, -4.7; and frontal, 7.0. Before beginning Brain electrical stimulation, a minimum one week must pass. once each rat’s discharge threshold has been established. Increased stimuli following discharges are used to measure seizure stage, duration, and amplitude of behavioural seizures.

Phytochemicals responsible for Anticonvulsant Activity.^54-60

Flavonoids

These polyphenolic compounds, found in fruits, vegetables, and herbs, have demonstrated antioxidant, anti-inflammatory, and neuroprotective properties. Flavonoids like quercetin, kaempferol, and luteolin have shown promise in reducing seizure frequency and protecting against neuronal damage.

Terpenoids

These are a large and diverse group of naturally occurring compounds found in essential oils of various plants. Linalool, for instance, found in lavender, has been studied for its sedative and anti-seizure properties.

Alkaloids

Plant-derived alkaloids such as pilocarpine (from Pilocarpus microphyllus) and corydine (from Corydalis species) have been found to exhibit anticonvulsant activity in animal models.

Phenolic Acids

Compounds like rosmarinic acid from rosemary have been researched for their antioxidant and anti-inflammatory effects, which could contribute to their anticonvulsant potential.

Cannabinoids

These are compounds found in the cannabis plant, such as CBD (cannabidiol), which has shown anticonvulsant effects, particularly in drug-resistant forms of epilepsy like Dravet syndrome.

Conclusion

Epilepsy continues to be a major worldwide health issue that impacts millions of people and presents complex challenges in its management. Despite advances in pharmacotherapy. substantial proportion of patients experience drug-resistant epilepsy, emphasizing the urgent need for novel and effective treatment strategies. The pathophysiology of epilepsy, rooted in aberrant electrical activity of the brain, involves dysregulation of ion channels and neurotransmitter systems such as GABA and glutamate pathways. These insights have guided the development of antiepileptic drugs (AEDs), which primarily target these mechanisms. However, the limitations of current AEDs, including side effects, high costs, and limited accessibility in low-income regions, necessitate the exploration of alternative therapeutic options.

Phytochemicals, derived from natural sources, offer a promising multitarget approach to epilepsy treatment. Preclinical studies highlight their ability to alter different pathways that are engaged in seizure genesis and propagation. However, further research is needed to clarify their mechanisms of action, determine optimal dosages, and ensure safety in human populations. With 80% of epilepsy cases concentrated in resource-limited settings, phytochemicals may provide cost-effective and accessible solutions. Moving forward, integrating phytochemical research into epilepsy management could transform therapeutic paradigms and improve patient outcomes, particularly in underserved populations, clearing the path for a more comprehensive inclusive method to epilepsy care.

Acknowledgement

The authors gratefully acknowledge the support and resources provided by Rajgad Dnyanpeeth’s College of Pharmacy, Bhor, which were instrumental in the completion of this review article.

Funding Sources

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Conflict of Interest

The authors declare that they have no conflict of interest.

Data Availability Statement

This statement does not apply to this article.

Ethics Statement

This research did not involve human participants, animal subjects, or any material that requires ethical approval.

Informed Consent Statement

This study did not involve human participants, and therefore, informed consent was not required.

Clinical Trial Registration

This research does not involve any clinical trials.

Permission to reproduce material from other sources

Not Applicable

Author contributions

Swapnali Jyotiba Bhagat: Conceptualization, Literature Review, Review & Editing.

Vishal Sudam Adak: Visualization

Shrikant Ramchandra Borate: Visualization

Rajkumar Virbhadrappa Shete: Supervision

Deepak Vitthalrao Fajage: Data Curation, Review & Editing

Shivkumar Manoharrao Sontakke: Project Administration

Jayshri Anna Tambe: Project Administration

Prajakta Anil Kakade: Project Administration

Reference

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Psoriasis is a long-standing autoimmune skin disease with predominant T-cell mediated inflammation leading to formation of erythematous plaques with silvery scale.¹ This disorder affects approximately 2–4% of the global population of world population, and although the incidences are not likely to differ considerably between different populations, geographical and ethnic disparities are observed.² For instance, Psoriasis is more frequently observed in Europeans and much less often in Asians and Africans. Apart from expressing itself as an unwelcome skin disease, psoriasis has a devastating effect on patients’ psychological and social wellbeing, quality of life. Sufferers often experience feelings of stigma, embarrassment and isolation which compounded the mental health problems posed by the disease which is for life. Also, psoriasis requires constant monitoring as it triggers time to time flare-up and time to time remission periods.³

However, many studies reveal that psoriasis is developed from various interactions of genetic background, environment and immune systems.⁴ Occasionally, there are genetic differences, including alleles in the major histocompatibility complex, more specifically, the HLA-Cw6 gene is strongly related to psoriasis.⁵ This indicates that predisposition to psoriasis, as signalled by these genes, is not sufficient to initiate the condition. Infection, stress, trauma, or some medications are examples of factors that provoke the condition and activate the disease process. These triggers mobilize the immune system, initiating inflammatory processes that in effect fuel the disease.⁶

New data in immunology have shown that certain cytokines play a central role in inflammatory responses involved in psoriasis.⁷ Pro-inflammatory cytokines, tumor necrosis factor-alpha (TNF-α), interleukin-17 (IL-17), and interleukin-23 (IL-23) have now been identified as central to the development of the disease.⁸ These cytokines sustain an inflammatory cycle by provoking Th17 cells and dendritic cells to promote keratinocyte hyperproliferation besides abnormally differentiating.⁹ This leads to the hallmark features of psoriasis: rash characterized by thick skin, scaled, and red in color. The molecular and cellular investigations into immunopathogenesis of psoriasis have not only advanced the scientific knowledge about the disease but have also opened new possibilities of its treatment. Other targeted therapies include TNF-α inhibitors such as infliximab and etanercept have significantly improved the clinical outcomes among patient with psoriasis; IL-17/ IL-23 antagonists: eculizumab and Ustekinumab have similar benefits among patient with psoriasis.¹⁰

However, these therapeutic continue to be faced by various challenges. Traditional organ-targeting agents and biologic mostly are ineffectual or lack long-term remission in all the patients. This implies that some therapies present negative effect such as immunosuppression, raised infection rate, and high cost hence not affordable by a large population of persons in the globe. For instance, the treatment of psoriasis is lifelong; the management of side effects of medications, as well as compliance to therapy regimens, is therefore a challenge. These considerations point to unmet medical needs that call for novel therapeutic interventions that can be more effective, safer and more widely available for the sickest of patients.¹¹

The systemic inflammation resulting from the psoriasis is also acknowledged to be related to co-morbid conditions of PsA, CVD, metabolic syndrome and depression. These co existing diseases aggravate the disease and add to the clinical and management challenges, underlining the need for a holistic and integrated model of health care delivery. New investigational drugs shed light on future research and treatment for psoriasis, but effective means to translate disease control to more patient-oriented outcome comparably remains a a complex clinical challenge in the management of the condition.^12,13

Current Treatment Landscape

The current options of managing Psoriasis are diverse since they focus on addressing the numerous violations of pathophysiology and signs in the course of the disease. Topical treatments including corticosteroids and Vitamin D derivatives like calcipotriol continue to be the recommended form of treatment for mild to moderate psoriasis. These agents have anti-inflammatory activity and can modality the proliferation of keratinocytes, mainly because of their poor penetrative profile for psoriatic plaques and the side effects including skin atrophy, the use of these agents is limited to long-term therapy. For more severe conditions, there is the use of photo therapy, including narrow band ultraviolet B and psoralen plus ultraviolet A are used where they can effectively relieve diseased symptoms through regulating the death of numerical keratinocyte cells and also modulating immunologic activity. Nevertheless, daily clinic visits and potential detrimental effects from long term skin exposure utilize the applicability of this technique.^14–16

Methotrexate as a systemic agent and cyclosporine and acitretin as other potent systemic agents are employed commonly for moderate to severe psoriasis.¹⁷ Although, these 689/. Biologic agents have become the areas of interest in managing psoriasis because of its focus on addressing particular immune processes. Biologics, including TNF-α inhibitors (etanercept, infliximab) and IL-17 inhibitors (eculizumab, afelimomab) and IL-23 inhibitors (guselkumab, tildrakizumab) yields high efficacy and long-term plaque control. However, due to expensive nature, risk of immunosuppression, and the requirement of parenteral administration they have limitations to use and compliance.¹⁸

Novel sensation, JAK inhibitors such as tofacitinib and TYK2 inhibitors like deucravacitinib have emerged as promising alternatives to biologics due to their oral administration and targeted cytokine inhibition. However, they may still cause systemic side effects and require appropriate monitoring and target specific cytokines.¹⁹ In addition, new strategies like RNA linked treatments, gene therapy and nanocarriers like Transethosomes are being introduced so that the drugs can be delivered effectively and achieve improved therapeutic effects with less side effects. However, current issues like high expense of biologics, lack of access in LMICs, and the long-term nature and flare of psoriasis indicate that IL-17 inhibition has room for growth. More so, as the knowledge on psoriasis advances, there will always be promise of incorporating newer and better therapies and drug delivery systems into practice to make the care more efficient and patient centered.²⁰

Transethosomes

Transethosomes are an improved vesicular formulation for the delivery of drugs and have been conceived as a very novel development in treatment of dermatological disorders especially psoriasis. These lipids based, highly flexible liposomes are developed to counter the problems posed by the SC, the outermost layer of the skin to deliver the drug to the deeper layer of the skin. Transethosomes are formed by phospholipids, ethanol and surfactants; this formulation improves both the flexibility and skin permeability when compared to ethosomes and transferosomes. Due to this formulation, transethosomes can carry from small molecules to biologics and other therapeutic agents effectively.²¹

Transethosomal SVs’ deformability can be regarded as the primary characteristic of this formulation, with ethanol and surfactants contributing to the effect. Ethanol weakens the intercellular lipid bilayer in the SC, which in turn enhances the permeability of the spaces and allows the LCS vesicles to travel through small channels.²² Surfactants increase the vesicular fluidity in a way that helps them change their shape and penetrate as far as the hyperkeratotic psoriatic plaques. There is much interest in this property because of the excessive thickening and inflammation of the skin layers in psoriasis that present a major challenge for the penetration of traditional topical treatments. They also afford the encapsulated drug from inhibition due to factors such as light, oxygen and enzymatic action. This is important in that the drugs will get to the target site in their intact and bioavailable form. Furthermore, the lipid bilayer of transethosomes exhibits the reservoir effect that allows the controlled and prolonged release of the drug. This means that it reduces the frequency of application and enhances patient compliance which are important considerations in managing chronic illnesses such as psoriasis.²³

Transethosomes can add both hydrophilic and lipophilic drug substances and thus are appropriate for the majority of potential therapeutic agents. Topical methotrexate, calcipotriol and corticosteroids in transethosomal formulations are proven to be very promising in treating psoriasis.²⁴ These formulations improve topical distribution of the drugs to the affected keratinocytes in psoriasis plaques and reduce side effects such as hepatotoxicity and skin atrophy. However, there is potential interest in using transethosomes for delivery of new generation therapeutic agents such as biologics, immunomodulators and natural compounds like curcumin, and resveratrol, which have ant-inflammatory and antioxidant activities. The use of transethosomes is not confined to psoriasis only but has scope in other dermatophytic disorders and in delivery of some systemic drugs through transdermal paths. But their synthesis is associated with issues like formulation site definition, stability of formulations when stored in appropriate containers, and how it can be scaled up for mass production. Still, these drawbacks have not deterred research because improvements to transethosomal technology are being made periodically and the therapeutic applications of this technology are expanding.²⁵

Mechanisms of Transethosomes in Psoriasis

Transethosomes are a newly synthesised drug delivery system that effectively counters some of the main problems of psoriasis treatment since they can deliver the drug to the psoriatic plaque. Due to the presence of phospholipids, ethanol, and surfactants, preparations of the corresponding group can easily cross the stratum corneum, a major barrier to most topical drugs. These highly deformable vesicles take advantage of the interactive possibilities of ethanol and surfactants to increase in flexibility and skin permeability. Ethanol makes the lipids in the stratum corneum less compact and permeable, while surfactants reduce density of vesicles thereby making it easier for transethosomes to pass through the tight intercellular spaces and even through thickened psoriatic skin.²⁶

Once transethosomes arrive at the targeted site, their lipid bilayer serves as carriers for the release of the entrapped drugs on a time-release basis. This localized delivery affords maximum drug concentrations at or near the site of inflammation and minimal systemic drug exposure and hence less dosage frequency is required. Furthermore, transethosomes help in shielding encapsulated drugs from different degradative processes such as oxidation and enzymatic activity thus resulting in increased drug stability and bioavailability. This feature is particularly beneficial in handling of delicate therapeutic compounds such as biologics, natural bioactive, and small-molecule inhibitors all of which have found useful application in the treatment of psoriasis.²⁷

Transethosomes have a particularly higher value in controlling the cytokines that are main inflammatory aspects of psoriasis such as TNF-α, IL-17, and IL-23. The immune response-related targets can be reached at the site of skin affection due to the ability of transethosomes to deliver therapeutic agents in targeted and non-invasive manner. Moreover, capability to encapsulate both hydrophilic and lipophilic drugs makes these carriers even more universal, in particular, as for drug delivery systems, with the possibility of co-delivery of two drugs acting through different mechanisms. For instance, methotrexate and corticosteroids used individually can be put together in a single puff and used to treat both inflammation and keratinocyte hyperproliferation.²⁸

In psoriasis, the thickness of the hornified layer, the SC, which is furthermore lipid changed, is an enormous problem for conventional treatments. Transethosomes can easily overcome this barrier because of the deformability and the lipid like nature of the structure that enables them to penetrate deep into the psoriatic plaques. This capability in conjunction with long circulating times and capability to encapsulate a variety of drugs makes treatment regimens easier and patient compliance is enhanced. With unique advantages in overcoming major deficiencies of conventional therapeutics for Psoriasis, transethosomes demonstrate the prospects of topical, effective and comfortable approach to patients.²⁹

Advantages of Transethosomes

Transethosomes have several advantageous features that make them a fairly promising drug delivery system, for example in the treatment of psoriasis. These advantages are due to the structural, compositional, and functional features of these nanocarriers that allow the complication of the limitations of the old-fashioned delivery systems.

Enhanced Skin Penetration

Free-form transethosomes are formulated with the highly flexible liposomes that are capable to transport active molecules even in the high-density keratinized part of the epidermis found in psoriatic plaques. The transport advantage comes from their flexible bilayer structure that is physically loaded with ethanol and surfactants that enable the NPs to slip through the histological intercellular spaces for efficient drug delivery to the target tissues. This improved access guarantees that the therapeutic agents get to their site of action than the common preparations.³⁰

Improved Drug Bioavailability

The transethosomal system, which includes both hydrophilic and lipophilic drugs, can greatly enhance the efficacy of a large number of pharmaceuticals. The vesicles provide shelter to the encapsulated drugs from the effects resulting from external conditions such as light, oxygen and enzymes. This protective effect means that more of the active drug floods the target site thereby improving the therapeutic effectiveness.³¹

Localized Delivery and Reduced Systemic Toxicity

The transethosomal approach delivers drug in a localized form and targets the affected area of skin, in this case, psoriasis. This proven approach is least chances of becoming systemic, cutting out potential systemic side effects probably known with methotrexate, corticosteroids and immunomodulators. For instance, methotrexate –loaded transethosomes exhibited good plaque reduction with lowest cumulative exposure to the systemic circulation, thereby is safer for long-term treatments.³²

Sustained Drug Release

Encapsulation of drugs is achieved within the lipid bilayer of transethosomes thereby allowing the process will be continuous over a very long time. This sustained release will give a longer period for the drug to work and consequently have few applications but better patients’ compliance. This feature becomes most helpful when dealing with diseases such as psoriasis where constant drug levels are most important due to the persistence of the disease.³³

Versatility in Drug Loading

Transethosomes can contain wide range of molecules such as small molecular weight drugs, biologics, natural substances, and even nucleic acids. Because of this flexibility, they may be used in any form of treatment, including anti-inflammatory agents and immunomodulators, siRNA, and phytochemicals. Their ability to delivery methotrexate together with calcipotriol as a combination therapy widens their therapeutic value in eliciting an effect on different pathways involved in disease pathophysiology.³⁴

Reduced Skin Irritation

Transethosomes do not only increase vesicle deformability through the incorporation of ethanol and surfactants but also reduce skin sensitivity through the reduction of free drug molecules in contact with skin. This property has made transethosomal formulations more acceptable to the patients especially those having sensitive or inflamed skin such as in psoriasis cases.³⁵

Ease of Application and Patient Compliance

The topical formulations are mainly presented as gels and creams and the transethosomal formulations are easy to apply topically. The fact that they are able to deliver a constant stream of drug makes repeated application not necessary especially for patients that need long term treatment. This convenience improves on patient compliance a very important aspect in the management of chronic illnesses.³⁶

Improved Stability of Therapeutic Agents

The structure of transethosomes involves a lipid bilayer layer that protects the encapsulated drugs from some factors such as light, oxygen and enzymatic action. This enhanced stability increases the shelf life of the formulation, where by the drug does not spoil before the intended application time.³⁷

Potential for Multi-Target Therapy

Transethosomes are excellent in multi-drug delivery systems in which more than one therapeutic agent can be incorporated, and each of them possesses a different mode of action. This is especially useful in chronic diseases such as psoriasis, where an optimal anti-inflammatory, immunosuppressive, and keratinocyte modulation drug regimen is superior to individual treatment.³⁸

Adaptability Across Routes of Administration

While the transethosomes can be primarily applied on the skin surface they can be useful for other routes of administration as well, including transdermal, nasal and oral. This versatility opens up other therapy areas for the platforms increasing the value and versatility of the drug delivery systems.³⁹

Superior Performance Compared to Other Formulations

Transethosomes exhibit multiple advantages over other topical drug delivery systems, such as liposomes, nanoemulsions, and traditional hydrogels. Table 1 provides a comparative overview demonstrating the superior attributes of transethosomal formulations relative to other topical systems.

Table 1: Comparative Performance of Topical Formulations of transethosomes in Psoriasis Management

Challenges in the Development of Transethosomes

As with other novel drug delivery systems, the prospects for transforming transethosomes into a highly effective treatment for psoriasis are accompanied by numerous issues that need to be solved in order to fully demonstrate their therapeutic advantages and substantial market value. These challenges affect many facets of development, such as formulation science, manufacturing techniques, stability, regulatory issues and patient availability. Challenges regarding the formulation are shown in Table 2.

Formulation Complexity

Transethosomes are elaborate structures that consist of multi-component comprising the phospholipid layer, ethanol, surfactant, and therapeutic API. This again means that the various components need to be designed in a manner that enhances stability, drug loading, with other characteristics of a controlled release system. Nonetheless, any change in the composition can distort the shape of the vesicles, cluster them, cause leakage, or minimize their effectiveness. Furthermore, adapting these formulations to encompass all the classes of therapeutic agents starting from small molecules to biologics makes the development process even more challenging. For instance, the hydrophilic and lipophilic medications pose different challenges with regards to the vesicular structure so that they can be formulated appropriately.⁴⁵

Scalability and Manufacturing Consistency

Transethosomal formulations have been studied in vitro and in small scale production; however, their large-scale synthesis in an industrial manner is not easy. Techniques like high pressure homogenization, ultrasonication or methods like thin-film hydration can be a challenge to standardize while maintaining cost constraint and quality control in large scale production so while the batches are being standardized errors should not rise. Keeping production repeats consistent in multiple production emailing is a topic of paramount concern for regulatory requirements and market acceptance. Furthermore, the pieces of equipment needed to facilitate some of these processes, such as encapsulation and emulsion, are costly and not easily accessible thus making the transethosomal formulations less available for wider use in clinical diagnosis.⁴⁶

Stability and Shelf Life

A stability factor is among the basics of transferring and developing formulations in a transethosomal manner. A principal product, ethanol, is volatile and may disappear hence destabilizing the formation of vesicles. The next required ingredient, phospholipids, is sensitive to oxidation and hydrolysis that can lead to vesicle breakdown and deco! From this perspective, both liposomes and reverse micronized formulations remain comparable. These stability problems are known to worsen with other factors that include temperature, humidity and light among others. In response to those challenges, methods like ‘lyophilization’ (or ‘freeze-drying’), use of ‘cryoprotectants’, and incorporating anti-oxidants need to be formulated. However, these additional processes are actual additions to the production process and can contribute to its complication and, respectively, to the increase in its costs.⁴⁷

Skin Penetration and Retention

The trans epidermal delivery of therapeutic agents to psoriatic plaques is also considered a major challenge. The epidermal pathology in psoriasis entails hyperkeratosis, which may affect the permeability of the skin to the penetration of the PTVs even when made more fluid by embedding in stereoisomers. More work is required to standardize and increase the effectiveness and penetration of the drug to the deeper skin layers by optimizing the size and surfaces charge of the P-VEs, their elasticity, as well as efficiency of drug entrapment into the P-VEs. In addition, achieving long-lasting reservoir at the site of action is considered a critical requisite for exponential therapeutic benefits. This include making the formulation to be non-digested quickly while at the same being available at the site of action.⁴⁸

Regulatory and Safety Concerns

It has become possible to transmit active substances through the use of ethanol and surfactants within transethosomal formulations using the current available formula, and these may lead to skin irritation, allergic reactions, or other extended toxicity. Although these components improve the capacity of flexibility and permeation of transethosomes, repeated use of these components as topical products in irritation-sensitive regions like psoriatic lesion calls for thorough safety studies. For transethosomal formulations, the standard and rigorous approval procedures are typical since transethosomal formulations are novel with unique characteristics: safety, efficacy and critical quality cannot be fully assessed by conventional measurements, so numerous preclinical and clinical data must be provided. The absence of uniform regulatory requirements to such sophisticated delivery systems adds to it the difficulties of the approval process.⁵⁰

Cost and Accessibility

The cost of the raw materials such as phospholipids and ethanol used in the preparation of the transethosomal formulations, the complexity of the machinery used, and the rigorous preparation methods has the effect of making transethosomal formulations significantly more costly than traditional treatments. These costs help define who has access to them and although cheaper compared to private sector technology solutions, may not be cheap enough to allow for its general use by patient population in low-income countries or for most patients in the developed world. Reducing the production cost and aiming at the possibility of utilizing cheaper source materials be other major solutions that will be very vital in achieving this accessibility. Moreover, clear differentiation of the new systems’ efficacy over other systems and other therapeutic approaches will also be important to explain the higher costs of these new and more complex formulations.⁴⁹

Patient Compliance and Acceptance

Between them, both technologies are appreciated for their ability to bring patient compliance, which is a basic factor in any kind of drug delivery system. Although transethosomal formulations are designed to ensure improved therapeutic efficacy, their application by patients also deserves consideration. This suggests that things like the ease of application, the how often one has to apply the product, and how unpleasant the feel of the product is, do play a role in level of compliance with the treatment. Processing solutions have to be developed with specific consideration of the targeted consumer, taking into account the convenience and tolerability of the formulations⁵⁰

Table 2: Stability Challenges in Transethosomal Gels with Potential Solutions

Overview of Clinical Studies

There are clinical studies done on transethosomal formulations for psoriasis which have pointed that this type of treatment is going to change the concept of treatment for this system, due to its targeted delivery and vesicular design, has shown promise in enhancing drug localization and minimizing adverse effects in preliminary studies. For example, methotrexate incorporated in transethosomes has shown increased localization of the drug-loaded system within the psoriatic plaque, resulting in decreased inflammation and severity of the plaques at lower overall doses of the drug. Consequently, this targeted delivery minimizes toxicity at the organismic level, and transethosomal methotrexate can, therefore, be used in long-term management. Likewise, transethosomal calcipotriol has enhanced biopharmaceutical properties and therapeutic efficacy over standard formulations, supporting faster reversion of the lesion-containing keratinocytes and sustaining restoration at a longer interval. Topical preparations including betamethasone, incorporated in transethosomal gels have also demonstrated increased anti-inflammatory activities with reduced dosages consequently reducing skin atrophy and hormonal imbalance. Tacrolimus and cyclosporine – two immunomodulators, which are employed for treatment-resistant psoriasis – were proven to be effective when delivered through transethosomes. These formulations have high efficacy of lesion clearance due to their action on psoriatic plaques and low general immunosuppression and the related hazards.  In recent years, clinical reviews have focused on the covalent therapy of methotrexate with calcipotriol in transethosomal systems because of their summate action. In addition, some initial explorations of biologics and herbal extracts incorporated in transethosomes have shown the possibility of new and easy to use therapies. These studies indicate the benefits of transethosomes but more large, long-term investigations are required to determine their clinical use and applicability in a variety of population subgroups.^54–56

Specific Drugs in Transethosomal Formulations

Cyards of transethosomal formulations have been utilized to improve the therapeutic efficiency of several drugs applied in the treatment of psoriasis while reducing their system toxicity. Oral methotrexate is one of the most effective systemic treatments for psoriasis, but its use is accompanied by substantial systemic side effects. Methotrexate does not exert its anti-psoriatic effect when administered orally but when formulated in transethosomal delivery system, it causes localized action at psoriatic plaques minimizing inflammation with a correspondingly diminished systemic exposure and toxicity. This targeted approach procures the advantage of improving patients’ safety and expanding its clinical applicability. Calcipotriol, a vitamin D derivative, an important regulatory intracellular mediator of the keratinocyte proliferation/differentiation process. However, the use of calcipotriol in the treatment of psoriasis is still limited because the drug penetrates poorly into psoriatic plaques. Transethosomal gels are preferred over other traditional systems because they increase the bioavailability of the delivery system with increased penetration depth thereby increasing its benefits to the patient. In the same way, the transethosomal encapsulation positively impacts corticosteroids such as betamethasone which are famous due to their high anti-inflammatory activity. These formulations allow targeted application, which in essence helps to decrease inflammation in the area of psoriatic lesions and, at the same time, minimize such adverse effects of long-term corticosteroid therapy as local skin atrophy and distant effects.

In the treatment of refractory psoriasis, immunomodulators such as tacrolimus and cyclosporine are used often. The use of these topical drugs in transethosomal formulations also promotes penetration of the drugs directly to the psoriatic plaques thereby augmenting therapeutic effects while reducing immunosuppressive action and hence accompanying side effects. This targeted delivery mechanism means these drugs are more appropriate for long term use in severe psoriasis conditions. New molecular entities, novel formulation containing biologics and or herbs and plant extracts and combinations such as methotrexate and calcipotriol have also demonstrated promising outcomes in experimental models. These new formulations do more than add to the many treatment options for psoriasis; they also open the gates for an individual and simultaneous approach to the condition. These technologies therefore showcase transethosomal formulations as a revolutionary improvement to the treatment of Psoriasis, as the drugs highlighted preceding presents a marked enhance in delivery and efficacy over the normally utilised treatments.^57–59 Key Studies on Transethosomal Gels in Psoriasis are shown in Table 3.

Table 3: Key Studies on Transethosomal Gels in Psoriasis

Future Directions

The future of the transethosomes in the psoriasis therapy is bright by the prospects made in formulation techniques, therapeutic substances and developing technologies. One of the most promising trends implies the creation of sophisticated multi-component transethosomal systems, for example, containing ligands selectively binding to targets characteristic of psoriasis, including IL-17 or TNF-α. Many of these targeted systems will improve the efficacy of drug delivery while at the same time reducing side effects. Further, the topical multi-drug delivery systems conjugated with anti-inflammatory agents covalently with biologics or APP’s small-molecule inhibitors could also minimize the complexity of this pathophysiology in psoriasis. Based on genomics and proteomics, the concept of personalized medicine will also progress the development of transethosomal formulations specific for patient characteristics to produce the best therapeutic outcomes with least side effects.^65–67

The new compound that can be used therapeutically also creates new avenues for transethosomal delivery. The administration of small interfering RNA (siRNA) strategy that targets inflammatory cytokine like IL-23 is a new promising strategy to treat inflammation at the gene level. Like with other curcumin or resveratrol based phytochemicals and natural products, encapsulation in transethosome brings a natural low toxicity formulation for the control of chronic inflammation of psoriasis ⁶⁸. Moreover, approaches to the drug delivery through the skin other than topical, e.g. via microneedle system or by using systemic applications might significantly increase the drug uptake and efficacy particularly in cases where the large surface of affected skin is covered or psoriatic arthritis (PA) is involved.⁶⁹

Therefore, further long term studies into the efficacy and safety of transethosomal nanocarriers would be important as part of strategies to successfully incorporate the transethosomes into clinical practice.⁷⁰ Consequently, chronic use and age-related studies for paediatric as well as elderly patients will confirm their efficacy among different population subgroups. The improvements in stability, drug loading capacity and release kinetics resulting from nanotechnology and hybrid systems will be reinforced by the integration of emergent technologies.⁷¹ Application of artificial intelligence and machine learning may have future for the prediction of the best formulation parameters and the individual therapeutic regimens in patients, 3D technologies may help to create personalized transethosomal delivery systems.⁷²

Thus, both, regulatory and commercial factors these are critical to the future of transethosomes. Accomplishing consistent characterization of transethosomes and researching efficient, low-cost large-scale production methods will provide an appropriate backdrop for the receipt of favorable regulatory responses for the commercialization of transethosomes.^73,74 This will alert the overall healthcare populace and especially the customers, which are the healthcare professionals and patients about the advantage of installing and implementing these enhanced systems in their working environment. Moreover, it would be advisable to integrate transethosome based therapies with other complementary therapies, including phototherapy or qualitative life style approaches, which could add extra value to the treatment of psoriasis. These advancements combined make transethosomes as a prompt solution to treat patient with psoriasis.⁷⁵

Novel Therapeutic Agents

The incorporation of new therapeutic agents into transethosomes is an innovation in the therapeutic management of psoriasis since transethosomes are vesicular carriers with unique characteristics that enable improved outcomes and selectivity ⁷⁶. A major development in this field is the Small interfering RNA or siRNA that is produced to target condition related genes including IL-17, IL-23 or TNF- α. Through neutralization of these important activators of psoriatic inflammation, transethosomal delivery of siRNA provides a targeted and localized knockdowns ensuring no undesired system-wide effects, which gives a tremendous advance in molecular targeting of the disease.⁷⁷

More therapeutic agents that can be encapsulated with transethosomal system include biologics, which include monoclonal antibodies, and fusion proteins.⁷⁸ Commonly these large molecules have poor depth of skin penetration across the layer of dead cells called the stratum corneum particularly when used in topical systems. Transethosomal encapsulation of biologics is useful not only for enhancing penetration into psoriatic tissues but also for delivering the drug selectively to the site of inflammation to reduce toxicity problems associated with systemic bioavailability.⁷⁹ Likewise, Smothered small-molecule inhibitors of intracellular signaling pathways like JAK-STAT or NF-κB activation can also be incorporated into transethosomes with optimal set­tings for sustained release and maintained therapeutic concentration at the inflamed loci.⁸⁰

Anti-inflammatory and antioxidant bioactive and phytochemicals such as curcumin, resveratrol and quercetin, meet numerous issues related to solubility and accessibility.⁸¹ Because these agents can be stabilised and perhaps enhance their therapeutic profile when incorporated with the transethosomes, they are considered suitable for the treatment of psoriasis with fewer side effects than synthetic drugs. Also, peptide-based drugs and nucleic acids-based treatment like antisense oligonucleotides, which are bioavailability restricted due to extensive enzymatic degradation can be delivered efficiently by transethosomes. This protective encapsulation guarantees these innovative therapies act at the target site as a deliverer of therapeutic concentrations.⁸²

Conclusion

Therefore, psoriasis is a proved to be a tough chronic inflammatory disease which still poses immense difficulties to manage and treat due the constrains of current treatment options. Recent lipid-based vesicular carriers, transethosomes, provide an encouraging solution, since they increase drug permeation, stability, and the delivery of the therapeutic products to the site of the psoriatic plaques, rather than the overall body surface, reducing side effects and boosting patient compliance. Bioavailability, stability and the ability to deliver various therapeutic agents such as biologics, small molecules or phytochemicals in drug delivery systems make them apt to transform psoriasis therapy. However, formulation issues, scale-up, and regulatory approval remain the main barriers to the development of transethosomal systems; On the other hand, ever-growing concerns on nanotechnology, personalized therapy, and pharmacogenomics are the increased possibilities of improving transethosomal systems. To develop these new delivery systems further, more extensive clinical trials, more robust stability profiles, and cheaper manufacturing processes should be investigated by future research to bring these new systems into the centre of clinical practice and to make a long overdue, positive impact on the quality of life of sufferers of psoriasis around the globe.

Acknowledgement

The authors are truly thankful to PRES’s College of Pharmacy (For Women), Chincholi, Nashik, for giving them the support and access they needed to prepare the manuscript. We are thankful to our mentors and colleagues for their ideas and support throughout the creation of this work. I am grateful to the library and IT staff for helping me retrieve literature and format my documents.

Funding Sources

The author(s) received no financial support for the research, authorship, and/or publication of this article.

Conflict of Interest

The authors do not have any conflict of interest.

Data Availability Statement

This statement does not apply to this article.

Ethics Statement

This research did not involve human participants, animal subjects, or any material that requires ethical approval.

Informed Consent Statement

This study did not involve human participants, and therefore, informed consent was not required.

Clinical Trial Registration

This research does not involve any clinical trials.

Permission to reproduce material from other sources

Not Applicable 

Author Contributions

Pooja Gajanan Akoshkar – Conceptualization , Project administration, Methodology, Writing original draft

Rahul Dnyaneshwar Khaire – Analysis, Visualization

Vikas Dhamu Kunde– Supervision, Reviewing

Pratiksha Suresh Katkade– Reviewing, Visualization

Gauravi Sunilrao Kherde – Analysis, Data Collection

Komal Sunil Taru – Reviewing

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In 1999, Anastas introduced the term “Green Analytical Chemistry,” describing it as the creation of analytical techniques aimed at reducing the production of hazardous waste. He emphasized the importance of environmental responsibility and pollution reduction in the design and evaluation of new analytical techniques.

Green Analytical Chemistry (GAC) is centered on creating eco-friendly analytical techniques by reducing the use of hazardous reagents, minimizing waste, and promoting sustainable chemical analysis. It originates from the broader field of “green chemistry,” which seeks to mitigate the environmental impact of chemical processes. GAC emphasizes several key strategies: replacing toxic reagents with safer alternatives, miniaturizing procedures to minimize the volume of chemicals used, and automating processes to enhance efficiency and reduce human exposure to hazardous substances.¹These strategies significantly reduce the consumption of reagents and the generation of waste, thus mitigating the environmental impact. Additionally, GAC promotes the integration of on-line decontamination or passivation techniques, which involve treating waste as it is generated to neutralize harmful substances before they can cause environmental harm. This approach not only makes analytical methods greener but also supports the broader goal of sustainable chemistry by ensuring that the entire lifecycle of an analytical process, from sample preparation to waste management, is environmentally responsible. GAC thus represents a crucial advancement in the effort to make chemical analysis more sustainable and less harmful to the planet² To align with sustainability standards, GAC offers extensive resources on green awareness, with a growing focus on green assessments over the past decade. This has included the development of various metrics to quantify the sustainability of methods. These tools not only guide scientists but also motivate them to adopt more environmentally responsible practices. The use of chemicals, energy use, sample volumes, extraction durations, and waste generation are among the important variables assessed. A number of reviews have recently looked at the suggested green metrics.^3,4

Green chemistry is a fundamental approach in chemistry that seeks to harmonize chemical processes with sustainable development principles. It encompasses various strategies aimed at reducing the environmental impact of chemical reactions and products. In the context of analytical chemistry, this discipline plays a vital role in promoting sustainable development, provided that analytical procedures are not only high-quality characterized by sensitivity, precision, and accuracy but also adhere to environmental sustainability principles. The present assay provides a succinct synopsis of the development of green chemistry and its multifarious impacts, encompassing its impact on analytical Chemistry, academic research, and education. The discussion also highlights the distinction between “sustainability” and “greenness” in this context. As a result, we provide at least foundational insights into sustainability within analytical chemistry. It is our hope that these reflections will inspire further advancements in both green analytical chemistry and sustainable analytical chemistry.⁵In recent years, a significant focus in analytical chemistry has been on advancing green analysis and adopting environmentally friendly approaches. This includes selecting safer procedures, solvents, and techniques, as well as employing analytical methods that produce less hazardous waste. Many chemical processes traditionally rely on solvents for purposes such as dissolution, extraction, purification, and as carriers or mobile phases, in addition to their role in enhancing spectral properties. Similarly, the use of chemical indicators, oxidants, and color development reagents is fundamental to numerous analytical procedures.

This review explores the adoption of green and eco-friendly methods, reagents, solvents, and techniques in the analytical determination of various analytes.⁶ Green chemistry refers to the implementation of strategies and techniques that minimize or eliminate the use of hazardous materials in production, products, byproducts, solvents, and reagents, thereby reducing risks to the environment and public health.^7,8 The design of materials or chemical processes is the main focus of green chemistry, with a special emphasis on four of the twelve important principles.⁹ All sciences, including chemistry and chemical engineering, are advancing rapidly, as evidenced by the increasing number of scientific publications and citations across various fields. Analytical chemistry is no exception, with modern methods offering unprecedented capabilities. Current technological and methodological advancements enable the detection of analytes at progressively lower concentrations, the separation of increasingly complex mixtures, and the achievement of higher precision and accuracy, all while requiring smaller sample sizes and improving analysis speed and simplicity. In addition to these developments, “green analytical chemistry “a term used to describe the emphasis on reducing new techniques’ negative environmental effects while boosting their safety is a major trend in analytical chemistry.^10,11

Green chemistry also concentrates on developing synthetic procedures and goods that reduce or do away with the usage of dangerous materials. Environmentally friendly chemistry includes all phases of the chemical life cycle, such as the development, application, and elimination of chemicals. Green chemicals are recycled or break down into innocuous residues. Hazardous materials in the surroundings can damage flora and fauna because of things like ozone layer depletion and global temperature change. exhaustion and the creation of haze.^12,13 The goal of green analytical chemistry (GAC) is to provide analytical techniques that are advantageous for analysts and the environment.^14,15Using the GAC technique has a number of benefits, such as using less toxic chemicals and reagents, using energy-efficient equipment, and producing less trash. One quick and very efficient chromatographic method HPLC. Smaller size and more accurate columns are features of HPLC systems, which can be very important in laboratory environments.^16,17 HPLC can handle any soluble substance, irrespective of its volatility. Choosing appropriate eco-friendly solvents can be facilitated by various solvent selection guidelines provided by different organizations.^18,19

The methodology involves systematically analyzing existing literature to highlight advancements in integrating Green Analytical Chemistry (GAC) principles with Quality by Design (QbD) approaches. The review begins by defining the role of GAC in minimizing environmental impact through the use of eco-friendly solvents, miniaturization techniques, and waste-reduction strategies. It then explores QbD principles, emphasizing Analytical Target Profile (ATP), risk assessment (Ishikawa diagrams), and optimization using Design of Experiments (DoE) to ensure method robustness and sustainability. The article also evaluates different greenness assessment tools, such as GAPI, NEMI, Analytical Eco-Scale, and AGREE, providing comparative insights into their effectiveness in quantifying the environmental impact of analytical methods. Additionally, case studies from various industries (pharmaceutical, food, and environmental analysis) are reviewed to demonstrate the practical implementation of these concepts. The methodology concludes by identifying challenges, future research directions, and the need for regulatory alignment, offering a comprehensive perspective on the evolution of green and QbD-based analytical method development.

Quality by design

Analytical chemistry is primarily used by pharmacists and chemical scientists in the industrial sector as an instrument for measurement to monitor the chemical reactions and ensure that technology and products meet high standards. Process Analytical Technology (PAT) focuses on monitoring and controlling analytical chemistry processes to support problem-solving and product composition analysis. The goal is to develop and oversee procedures that result in better final products a strategy known as QbD. The drug industry sets the standard for applying quality by design, to process development, and this has a direct bearing on the PAT methodologies and analytical procedures used. ²⁰

The primary objective of Green Chemistry is to reduce or eliminate the harmful effects of various organic solvents on both the environment and human health. Analytical methods typically balance environmental considerations with safety. By integrating sustainable chemistry with Quality by Design (QbD) principles, it is possible to develop techniques that are both dependable and environmentally friendly.²¹

Green Chemistry was founded with the goal of minimizing or doing away with the harmful impact that organic solvents have on the environment and human health.²² Conventional analytical methods frequently sacrifice environmental impact and safety. GAC principles can be combined with quality by design to build durable and environmentally friendly techniques. The importance and advantages of integrating QbD and GAC in analytical methods are emphasized in this review. In order to obtain optimal performance, QbD offers a methodical and systematic approach to method development in pharmaceutical analysis^.23 It centres on a comprehensive assessment and investigation of various approaches. Green sample preparation, waste minimization, the reduction in size of tools for analysis, and efficient waste treatment techniques are important facets of Green Analytical Chemistry.²⁴ Analytical methods’ environmental friendliness is evaluated using tools like solvent selection recommendations, the AMVI methodology, and the HPLC-EAT. A vital statistical tool in QbD, experiment design, is essential to this procedure. To sum up, the integration of Green Analytical Chemistry with QbD yields the advantages of both strong methods and sustainable environmental practices. Economic benefits have also been shown by applying similar ideas to High Performance liquid chromatography and other bioanalytical techniques. But more research and development are required. Before a product is finished, quality and robustness are ensured through the use of QbD, which is becoming more and more significant.^.25

Green Analytical Chemistry

The Green Analytical Chemistry journal aims to spotlight eco-friendly approaches that are often underrepresented in other analytical chemistry publications but are growing in significance. The principal aim of the magazine is to curtail or eradicate the utilization of detrimental agents and mitigate the production of waste. Also seeks to promote the use of screening methods for simple qualitative analysis, designed to bypass the extensive sample processing typically required for comprehensive quantitative analysis in large laboratories, replacing these with on-site and in-vivo technologies.

Green is the colour linked to money and chlorophyll. Environmental activists have been fighting hard in recent years to promote sustainability, and “green” product marketing has become popular. The adoption and application of green chemistry principles has become imperative for chemists in all domains of the chemical sciences, encompassing fundamental and practical research, manufacturing, and instruction.²⁶

The primary objective is to progress green analytical technologies’ foundational and applied aspects. The magazine will include both cutting-edge green analytical chemistry techniques and modified conventional methods, with a focus on technology appropriate for screening applications.^27,28 These techniques’ effects on the environment will be thoroughly evaluated in light of factors like reagent toxicity, waste generation, energy usage, and user safety. Additionally, the journal will provide a thorough evaluation of each method’s balance between environmental sustainability and functionality, taking into account both analytical and practical considerations. Published research will aid in advancing green regulatory approaches in analytical chemistry, ultimately minimizing the environmental impact of human activities. The content will also support university educators in integrating Green Analytical Chemistry into their curricula.^29,30

Greening Chromatography

Numerous articles discuss how chromatography can be made more environmentally friendly by utilizing smaller separation devices. or green solvents for e.g. Water likewise, “older” separation methods like gas chromatography (GC) and capillary electrophoresis (CE) are actually highly environmentally friendly methods. From a green chemistry perspective, these methods are advantageous when they are applicable and workable. However, CE and GC are not suitable for all compounds or complex sample matrices. Miniaturization offers a viable solution, with significant advancements being made in microfluidics and lab-on-a-chip technologies.^31,32

Greening Detection

In analytical chemistry detection, many greening efforts have focused on in situ analytical techniques, particularly spectroscopic methods.³³ From a green chemistry perspective, in situ analysis is beneficial as it enables real-time process monitoring and direct observation of hazardous or unsafe chemical synthesis. Additionally, it eliminates extra handling steps, which often lead to increased chemical and energy consumption. An example of such a technique is Raman spectroscopy, which has been integrated with microfluidic separation processes.³⁴ Due to its ability to penetrate thick sapphire glass windows, Raman spectroscopy is well-suited for in situ analysis in high-pressure fluid systems.^35,36 For instance, research has demonstrated that Raman spectroscopy can be used to determine solubility in SC-CO2. The application of Raman spectroscopy in processes, as an in-situ monitoring detector, for surface examination, and for in vivo biological system study demonstrated significant promise.

Green Development

To address society’s need for extensive information on the environment, products, and processes, much of the research in Analytical Chemistry relies on highly computerized methods. These methods incorporate various sample treatment and component separation techniques or utilize advanced detection technologies. One fundamental principle of green chemistry is the conservation of energy and materials through the integration of high-throughput systems. Another key aspect is the use of innovative materials, such as nanoparticles, microfilms, and micro-assays, which can significantly reduce waste and minimize the sample size required for analysis. The application of computing and communication technologies in chemical data analysis, particularly in sensor research and data management, streamlines the analytical process by reducing steps and energy consumption, aligning with green chemistry principles. However, despite these advancements, green chemistry remains underrepresented in analytical chemistry literature. The systematic incorporation of green chemistry principles into the selection of analytical methods for chemical data acquisition is still not well-established. Given the numerous books and review articles published on this subject in recent years, an increase in the development and refinement of analytical methods that adhere to green chemistry principles can be expected.^37,38

Sample preparation and separation science are the areas of analytical chemistry that are most commonly discussed in relation to green chemistry since they use the greatest number of solvents and other reagents. As a result, recycling, replacing hazardous solvents, and using solvents sparingly are given top attention. The “3R” method stands for reduction, replacement, and recycling.³⁹ Recycling, swapping out dangerous solvents, and utilizing solvents sparingly are therefore given great priority. The “3R” approach—Reduction, Replacement, and Recycling has driven the broader adoption of alternative solvents and the modernization of instrumental techniques. Additionally, it significantly impacts costs in large-scale analytical control laboratories.

Here are some examples:

R1-Reduction

The properties of solid supports and separation media change at high temperatures, increasing effectiveness and reducing the requirement for solvents.

Apart from requiring certain temperature-resistant packing materials, this technique has drawbacks, such as the possibility of thermally labile chemicals degrading on the column and solubility issues with hydrophobic molecules. Utilizing shorter columns, higher pressures, and narrow-bore columns, which have smaller particles, significantly lowers solvent use and waste production⁴⁰

R2-Replacement

In preparative scale HPLC, supercritical CO₂ is comparatively frequently utilized for the separation of physiologically active chemicals. Finding a substitute for the environmentally damaging acetonitrile is also a focus of intense research, with ethanol showing some encouraging results. Certain organic solvents are available, such as lactate esters derived from biomass feedstock. The employment of approved procedures, which contain acetonitrile as a required component, is mandated by several regulatory agencies, which poses a drawback to the novel eluents. Although ionic liquids are currently very popular solvents, they are unlikely to be useful in chromatographic procedures; instead, they may be useful in sample preparation techniques like extraction.⁴¹

R3-Recycling

To cut down on waste production, the lab should, whenever feasible, acquire the required equipment and recycle its solvents. Sadly, the energy requirements and high cost of this make it anything but a fully green solution.

A “4S” strategy was put up by M. Koel and M. Kaljurand for green analytical chemistry.⁴²

S1-Specific Methods

Capillary electrophoresis (CE), which replaces HPLC in identical circumstances, is the best illustration of adopting alternative approaches to solve similar problems. Capillary electrophoretic techniques offer greater success than HPLC in various applications due to their flexibility, high-efficiency separations, reduced solvent usage, and faster analysis times. With CE, you can utilize a variety of detector types, chromatographic separation processes, and an infinite selection of separation medium (buffers). Systems for collecting CE samples can be used. Flow analysis is a well-known technique for addressing certain analytical issues. It can be used to develop batch processes that are unreliable and have a significant potential to reduce waste production and agent use. Using techniques that enable direct probe analysis with little sample preparation is another way to go with this strategy. In this context, several laser-based and optical techniques ought to be taken into account.⁴³ Furthermore, spectroscopic techniques enable great selectivity and sensitivity to be maintained with straightforward, portable equipment, enabling in-situ or in-vivo real-time process monitoring and analysis.

S2-Smaller Dimension

The amount of solvents and chemicals used in the measurement (detection) steps is significantly reduced when measurements are carried out on a tiny silicon platform where customized micro systems incorporate several chemical handling processes. These compact devices can efficiently integrate analyte electrophoretic separation. One key advantage of miniaturization is its suitability for applications in bioscience and nanoscience, where only small sample sizes are available—such as individual cells, nanoparticles, or even single molecules.⁴⁴ Additionally, miniaturization contributes to energy conservation. However, microchip systems require small-scale reactors, pumps, and valves, often facing connectivity challenges. To address these issues, the Lab-on-a-Valve (LOV) concept has emerged as an innovative solution for pressure-driven sampling on a miniature scale.⁴⁵ This system effectively manages chemical vapor formation using programmable flow, enables non-chromatographic speciation, and supports renewable micro-solid phase extraction. Moreover, it facilitates on-chip optical and electrochemical detection, including autosensing techniques. Manipulating droplets of liquid samples is another way to solve interface problems in microfluidics. Single droplets are comparable to micro-sample vessels that can be used as reactors, extraction vessels, transportation devices for further analysis, and flexible input devices in other techniques such as CE. One such instance is the transportation of the separated analytes within the CE capillary to the CE column, where they are gradually fractionated into droplets. It takes less than fifteen minutes to do this process.⁴⁶

S3-Similar Methods

There are occasions when it is possible to gather enough data on chemicals without pre-treating the samples, or when a device without any moving components can handle the entire procedure. For “labs-on-a-chip” prototypes, paper can be used as an inexpensive, flexible, renewable, and biodegradable substrate. This material is easily obtained and performs well in a range of chemical and biological tests. Because the material is permeable. After use, this “device” is easily disposed of.⁴⁷

There is currently a growing selection of lateral flow devices accessible; for example, the typical pregnancy test combines immunoassay testing with porous materials or membranes. Modern mobile phones can capture images of response spots on paper and transmit them to a personal computer via the internet for further analysis. Using basic free software, a calibration curve can be generated, allowing for the completion of quantitative analysis.^48,49

S4-Statistics

Mathematical manipulation of the test results could provide information about the Analyte Signals; that is, the output data could be used to mathematically deduce a signal related to a particular analyte. Chemometrics is a branch of science that analyses chemical measurement data to extract chemical information. It is the study of chemical science that applies mathematical and statistical methods to design or choose the best measurement procedures and experiment. A substitute for chemical data processing could be chemometrics, which allows the application of far less complicated measuring techniques that usually do not need sample pre-treatment and lead to faster analysis times. Typically, multivariate statistical methods are applied in this situation. This is an example of how near infrared spectroscopy was applied in conjunction with chemometrics as a fast way to determine if the specimen’s matrices were changed by exposure to mine tailings pollution or not compared to what would be expected from wild populations. It was possible to ascertain in this case how much the harvest season, the person’s place of origin, and the level of soil pollution affected their chemical profiles.⁵⁰ Another example uses the NIR system to simulate the concentration of hexahydro-1,3,5-trinitro-1,3-S-triazine in a propellant factory by combining it with quantitative chemometrics. A fibre-optic probe from the spectrometer was instantly inserted into the continuously agitated samples in order to record in situ spectra. The recommended NIR technique eliminated the need for chemical reagents and the waste produced during the whole prediction procedure. The results show how quickly, thoroughly, and effectively the near-infrared technology can eliminate potentially dangerous residues from standard analytical procedures. Additionally, it might be used to keep an eye on the amount of RDX used in the manufacture of solid propellants.⁵¹ The chemometric approach proves highly beneficial in combined methods like LC and IR due to its ability to enhance the signal-to-noise ratio, adjust background interference, eliminate unwanted signals and residual mobile phases, and resolve overlapping chromatographic peaks, even under challenging gradient conditions with low chromatographic resolution. ^52,53 Additionally, significant cost savings can be achieved by utilizing large datasets for analysis and visualization. Exploratory data analysis aids in drawing conclusions without requiring an exhaustive chemical examination of each analyte. In this context, measurement results from chromatographic, electrophoretic, or spectroscopic techniques can be compared to a fingerprint—a unique profile representing the complex molecular composition of the sample. ^54,55 Over time, the detailed information within a chemically complex sample can be further explored to identify individual components and uncover underlying data. ^56,57 Fingerprint analysis is widely used for managing spectroscopic and chromatographic data, proving to be a powerful and efficient tool for quality control and assessing the authenticity of herbal products.^58,59

Principles of green chemistry

Chart 1: Green chemistry Click here to view Chart

The principles of Green Chemistry have been widely embraced across industries, government policies, education, and technological advancements globally.⁶⁰ A central objective of the circular economy is to create a balance between economic growth, resource sustainability, and environmental protection. Green chemistry plays a crucial role in driving this transition by promoting sustainable practices within the chemical industry. Furthermore, it has a significant impact on the development of innovative approaches in various branches of chemistry, including analytical chemistry, where the principles of Green Analytical Chemistry (GAC) are applied. One of the major challenges lies in establishing a suitable method for evaluating the environmental impact of an analytical procedure, considering multiple factors. Several published strategies have been introduced to assess the ecological footprint of different analytical techniques. ⁶¹

The twelve principles of green chemistry were developed by EPA scientists Paul Anastas and John Warner and elaborated upon in their 1998 book, Green Chemistry: Theory and Practice. These principles focus on reducing or eliminating hazardous substances in the synthesis, production, and application of chemical products to minimize risks to both the environment and public health. While it is challenging to implement all twelve principles simultaneously in the design of a green chemistry process, efforts are made to incorporate as many as possible throughout different stages of synthesis.⁶²

Prevention

Dealing with waste after it has been produced is less sustainable and less effective than preventing waste development. trash prevention, which is not only better for the environment and human health but also more economical than handling and discarding trash once it is created, is the primary tenet of green chemistry.⁶³

The relevance of this theory becomes evident when one considers that the United States produces over twelve billion tons of solid waste every year, of which approximately 300 million tons are dangerous waste. The chemical sector accounts for 70% of this hazardous waste; toxic organic waste, particularly methanol and xylenes, is a significant contribution. Waste disposal costs account for 2.2 percent of the US GDP, and they are constantly increasing.

Harmful to both people and the environment, toxic organic waste is mostly produced during specific phases of chemical synthesis, which are commonly known as “dirty reactions.” These reactions involve the use of toxic reactants and solvents and are characterized by harsh conditions that lead to the formation of numerous toxic byproducts. Common reactions that produce such waste include halogenation, oxidation, alkylation, nitration, and sulfonation, all of which are widely used across various industrial sectors.⁶⁴

While the chemical industry and other manufacturers have historically overlooked waste prevention, the core focus of green chemistry is to minimize waste generation. However, completely eliminating waste is practically impossible, as no raw material can be entirely utilized. When waste is discarded, it represents a permanent loss of valuable resources in the production-consumption cycle. Therefore, any effort to reintegrate these materials into the cycle is economically beneficial. It’s critical to determine first whether the production of trash might be prevented, and, if otherwise, to come up with strategies to optimize the amount of trash generated during production that is reused or otherwise put to good use.

Managing or mitigating the effects of hazardous, toxic, explosive, bio-accumulative, and waste chemicals is considered more favorable than attempting to control or prevent their synthesis altogether.⁶⁵

Atomic economy

Design or conduct the chemical process in a manner that ensures the final product contains a significant amount of the reactants, minimizing the loss of raw materials.

In 1990, Barry Trost introduced the concept of synthetic efficiency known as Atom Economy (AE), also referred to as Atom Efficiency. The old Boots process technique of synthesis had a low economic efficiency, using only around 40% of the raw components. This was a major problem. A novel “green” ibuprofen manufacturing technique was discovered in the 1990s that required only 3 steps. With a yield of up to 99%, this novel technique produced almost perfect transformation of every intermediary ingredient into the final result, while enabling material regeneration and reuse throughout the process. Consequently, the procedure virtually minimized trash formation, exemplifying “green synthesis.⁶⁶

To determine the percentage of atomic efficiency, use the following formula: % of atomic efficiency = (Mr of the desired product/Mr of all reactants) x 100 ⁶⁷

Less Hazardous Synthesis

Is a cornerstone of green chemistry, aiming to minimize the inherent risks connected with chemical procedure. It advocates for the design and execution of chemical reactions that produce minimal or no hazardous substances, prioritizing safety throughout the lifecycle of the chemical process, from raw material selection to the handling of by-products and waste. Traditional chemical syntheses often involve toxic reagents, intermediates, or by-products that pose risks to human health and the eco-system. This risk can manifest as harmfulness, flammability, explosiveness, or ecological damage. The principle of less hazardous synthesis challenges chemists to rethink these processes, focusing on minimizing or eliminating such dangers.

One way to achieve less hazardous synthesis is by choosing safer reagents and solvents that are less toxic or corrosive. For instance, water or ethanol can be used as solvents instead of more harmful organic solvents like benzene or chloroform. Additionally, the development of milder response circumstances like lower heat and pressures can reduce the likelihood of generating hazardous by-products or requiring harsh chemicals.⁶⁸

Another approach is the use of alternative, greener catalysts that are less toxic and more selective, which can minimize unwanted side reactions that produce harmful substances. Moreover, Hazardous waste production can also be decreased by engineering reactions that are more efficient in terms of atom economy, meaning that a larger proportion of the starting elements are incorporated into the finished product. Ultimately, fewer hazardous chemical syntheses contribute to a safer working environment, lower regulatory and disposal costs, and a reduced environmental footprint. This principle underscores the broader goals of green chemistry to make chemical manufacturing not only more sustainable but also safer for all stakeholders involved.

This idea encourages the creation of synthetic processes that put the usage and production of materials with little or no toxicity to the environment and human health first. Biological enzymes can be used as a substitute for dangerous chemicals in industrial processes to increase efficiency and save costs.⁶⁹

EX.- Conceptually simple, Asahi Kasei’s new polycarbonate (PC) production method substitutes CO₂ for hazardous carbonyl dichloride (COCl₂). Additionally, this method does away with the need for dichloromethane (CH₃Cl₂) as a solvent. To make polycarbonate and ethylene glycol (C₂H₆O₂), the total reaction involves ethylene oxide (C₂H₄O), CO₂, and bisphenol-A (C₁₅H₁₆O₂).⁷⁰

Designing Safer Chemicals

One of the hardest problems in creating safer products and processes is reducing toxicity without sacrificing a product’s usefulness or efficacy. Toxicology and environmental science concepts must be understood in addition to a thorough understanding of chemistry to achieve this balance. Due to their efficacy, Highly Reactive substances are frequently used by chemists for molecular transformations. These substances may, however, also interact with unanticipated biological targets and have negative consequences on both the environment and human health. Without a thorough grasp of the relationship between a chemical’s structure and its hazards, even the most skilled chemist may lack the necessary tools to address this challenge.

Innovative approaches to chemical characterization are necessary for success in toxicology, as it is acknowledged that hazard is a defect in molecular design that needs to be fixed from the beginning. One important chemical attribute that must be identified, evaluated, and controlled as part of an all-encompassing, systems-based approach to chemical design is the inherent danger of elements and compounds.

Now is the ideal time to start working together as toxicologists and chemists to develop a truly holistic, multidisciplinary strategy that will train the future generation of scientists to make safer compounds with the help of cutting-edge curriculum innovations. Toxicology is developing quickly, using molecular biology to understand the mechanisms underlying toxicity. The basis for formulating design guidelines that chemists might use to direct their efforts in producing safer chemicals is provided by an understanding of these mechanisms. As we enter a new chapter, we are prepared to light the way for a world that is more secure, more nutritious.⁷¹

Safer Solvents and Auxiliaries

Auxiliary compounds (solutes, separating agents, etc.) should be used sparingly or not at all when possible and shouldn’t be hazardous. Chromatographic separations, which need the use of massive volumes of solvents, raise environmental problems due to pollution. Numerous conventional organic solvents are hazardous, combustible, and caustic, and recovering them usually necessitates costly, heavy on energy distillation that results in large losses. This emphasizes the necessity of creating solvents that are more environmentally friendly.

The principle of “Safer Solvents and Auxiliaries” suggests the synthesis procedure should reduce, and Auxiliary chemicals should be avoided wherever possible. If their use is necessary, these substances should be non-hazardous. The long-term viability of the process, employee safety, security during the process, and environmental protection should all be given top priority when choosing appropriate substitutes for traditional organic solvents in accordance with the principles of green chemistry. The best solvents should be easily handled, quickly recyclable, have minimal volatility, as well as stability both chemically and physically.72 Based on their applicability, conventional solvents can be categorized as suitable, usable, or undesirable [Table 1].

Table 1: Solvent selection is based on their intended uses

Replacing traditional organic solvents with recyclable alternatives, including ionic fluids, is currently a promising strategy.⁷³ These are the salts that, when left at room temp., stay liquid. Ionic liquids provide safer chemical processes since they have a low vapor pressure and do not evaporate easily like volatile organic chemicals do.⁷⁴

Design for Energy Efficiency

It is imperative to acknowledge the environmental and economic ramifications of energy requirements in chemical processes and to take steps to mitigate them. Whenever possible, synthetic operations should be carried out under room pressure and temperature. Several energy-saving techniques were developed in response to the 1973 oil crisis to maximize the effectiveness of each kilojoule utilized in manufacturing. Minimizing energy consumption requires adherence to the Principle of Energy Efficiency, commonly known as Design for Energy Efficiency.⁷⁵

For example, Growers of tomatoes benefit from a greenhouse that uses leftover vapor from another nearby chemical processing facility to produce ammonia. The CO₂ levels in these greenhouses are kept below 50%, which promotes plant growth. To further encourage the growth of tomato seeds, carbon dioxide that has been collected from greenhouse gases can be utilized as a bi-activator⁷⁶

Use Renewable Feedstock

Whenever possible, renewable raw materials should be used instead of non-renewable ones, both technically and financially. Using renewable feedstocks whenever possible is emphasized by the 7th principle of green chemistry. For example, it is usually more advantageous to use materials that are renewable rather than a variety of waste-producing plastics. The trend of creating biodegradable plastics is a result of this. In the food business, biodegradable packaging is becoming more and more significant due to various variables like worldwide agricultural and energy resource demand, political developments, and changes in legislation.⁷⁷

Utilizing renewable raw materials lowers the release of carbon dioxide and improves energy usage in the bioplastics manufacturing process. For instance, the Coca Cola makes bottles with blends of polyethylene that contain thirty percent, and Nature Works, an American firm, makes bottles made of lactic acid-containing polymer The fermentation of dextrose, which is normally made from maize starch, yields PLA; about 2.5 kg of corn are required to make 1 kg of lactic acid containing polymer.⁷⁸

Reduce derivatives

Minimizing or avoiding unnecessary derivatization is essential, including the use of blocking groups, protection and deprotection steps, or temporary modifications to physical or chemical processes. These steps often require additional reagents and can lead to waste generation. Reducing the usage of chemical derivatives is only core tenets of green chemistry. This principle suggests that, whenever possible, physicochemical reactions involving the preventing and releasing of groups during synthesis be avoided.⁷⁹

Catalyst

Higher atom economy processes can be made possible by the application of catalysts. Because catalysts themselves aren’t consumed by chemical reactions, they can be recycled again and don’t add to waste. They can make it possible to use reactions that, in ordinary circumstances, would not take place but also result in reduced waste.⁸⁰

To support environmental protection, the principle of catalysis advocates for the use of biodegradable catalysts. These catalysts help to reduce energy consumption, avoid organochlorine compounds, and minimize water usage or waste. Throughout this process, enzymes remain unchanged and can be used repeatedly. They do not alter the energy levels of the reactants. While biocatalysts offer advantages such as faster reaction rates and greater specificity compared to non-biological catalysts, they can be limited by heat sensitivity and stability issues.⁸¹ 

Design for Degradation

Organic pollutants, such as halogenated compounds, cannot break down and can build up in the environment. Replace these chemicals as much as possible with ones that break down more quickly in the presence of water, UV radiation, or microorganisms. ⁸²

Real-time Pollution Prevention

To ensure that they do not end up in the ecosystem, chemical goods should be designed to break down into innocuous compounds after they have served their intended purpose.

The principle of designing for degradation requires that chemical products be formulated to break down into environmentally harmless substances once their intended function is complete. Meeting this requirement can be achieved by adjusting technological parameters during process management and modifying auxiliary substances used at various stages of production. The goal is to minimize the creation of harmful by-products and maximize the recycling of waste materials for reuse in production⁸³

Keeping an eye on a chemical reaction while it’s happening can assist stop dangerous and polluting compounds from escaping owing to mishaps or unanticipated reactions. Real-time monitoring makes it possible to identify warning signals and take appropriate action to avert or control an incident before it happens.^84,85

Safer Chemistry for Accident Prevention

Working with chemicals is never completely risk-free. Nonetheless, risk can be reduced with effective hazard management. There are obvious connections between this concept and several other principles that address dangerous materials or goods. Processes should, if feasible, be structured to minimize risks in the event that eliminating exposure to hazards is not feasible.^86,87

The Comparison of green analytical chemistry Methodologies with Conventional analytical chemistry Methodologies:

During the 1990s, Paul Anastas contributed to several publications on green analytical chemistry, emphasizing its significance within the broader framework of green chemistry and highlighting the necessity of transitioning toward environmentally sustainable practices. ⁸⁸Specifically, Anastas highlighted its dual function in environmental conservation, stemming from the possibility that different analytical chemistry techniques could not only aid in the identification of potentially harmful environmental elements but also contribute to the accumulation of pollutants and other ecological issues.⁸⁹ It is crucial to work toward making analytical procedures more environmentally friendly while still taking into account their sensitivity, accuracy, and precision.

Steps in the Development of an Ecological Mind-set in Analytical Laboratories:

Table 2: The Development of Environmental Awareness in Analytical Laboratories.

The rise in analysis activity on unfavorable environmental elements and environmental samples resulted in an increase in analytical wastes and a noticeable shift in the attitude of laboratories regarding the effects of their residues. The evolution of environmental awareness in analytical laboratories is evident (Table No. 2), driven by concerns over the negative impact of increasing reagent use and waste generation. This shift highlights a strong connection between ecological consciousness and the advancements in chemistry at the end of the 20th century. In this context, the concept of Green Analytical Chemistry (GAC) has provided significant opportunities from both academic and industrial perspectives.⁹⁰

Goal Of Green Analytical Chemistry

Reduction of use of chemical elements

Proper waste management

Prevents pollution

Decrease in energy consumption

Avoid the release of toxic solvent

Minimize hazardous materials

Efficient utilization of raw materials

Quality by Design and Analytical Chemistry

In the business world, Analytical Chemistry is primarily used by chemists and chemical engineers as a measurement tool to track chemical technological processes, which is the primary method of technology and product quality control. Process Analytical Technology (PAT) incorporates analytical chemistry’s process monitoring and control features, primarily for troubleshooting and analyzing product composition. Its objective is to develop and maintain technological processes that ensure the production of high-quality goods. This approach aligns with the concept of Quality by Design (QbD). The pharmaceutical industry has been at the forefront of utilizing QbD to enhance technical processes, influencing both analytical methodologies and PAT. Literature suggests that combining the QbD approach with Risk Analysis and Design of Experiments aids in developing reliable chemical processes. These processes incorporate appropriate analytical monitoring and control measures to ensure product quality. ^90,91

A fishbone diagram can also be used to illustrate sustainable production, which calls for the integration of green elements into every stage of the process. [ Figure 1].

Figure 1: Ishikawa diagram for examining the impact of potential variables on the LC method based on pilot studies and prior understanding Click here to view Figure

The required resources for process sustainability and high-quality goods are shown in the figure. In this instance, all inputs materials, energy, and waste have a significant bearing on green chemistry and should be considered when creating procedures and end products to guarantee that they are suitable for the purposes for which they are designed.

These days, QbD is the primary focus of pharmaceutical development, and as a result, analytical chemistry is being thoroughly researched. Web of Science reports that the keywords “Analytical Quality by Design” are published in over 100 papers annually. It appears that the quality-by-design principle is increasingly being included into contemporary methods for process development in chemistry and pharmacy. First and foremost, for that, the evaluation of the intended product’s performance profile.⁹²

Analytical Reagents

Customers look to analytical chemistry to (1) solve problems relating to compound purity or process and product safety and (2) provide enough quantities of pertinent elements and molecules. Chemical information about the object under study, both qualitative and quantitative, should be obtained with adequate accuracy and precision. A measurement process’s output must provide information that allows a user to make technically sound judgments for a specified goal

It is essential to acknowledge and utilize the diverse range of analytical techniques, protocols, and tools available, aligning them with the specific objectives of the analysis. According to ICH Guidelines Q8(R2) on Quality by Design, this approach involves a systematic development process that starts with well-defined objectives, focusing on product and process understanding, as well as process control, all grounded in scientific principles and quality risk management. You should refer to this material while you construct the quality-by-design analytical method. The excerpt highlights how crucial it is to choose the analysis method and the analysis procedure’s parameters of performance to achieve the study’s particular objectives. Following the Recommended Practice entails a change from a Quality by Testing strategy, in which batches that are released into the market or the final product are tested, to a more fully understood and efficient production process that enables adjustments to operational parameters, enhances measurement reliability, and offers regulatory flexibility. This strategy is in line with Green Chemistry’s tenets, which support real-time analysis to stop pollution. The concepts of Green Chemistry are naturally supported by waste management and chemical toxicity reduction, even though they are not usually addressed in quality control systems.

A comprehensive review published by T. Tome et al.⁹³which references more than 70 research, emphasizes the expanding use of Quality by Design (QbD) in analytical evaluation procedures. The phrase “Analytical QbD” is emerging in the field of analytical chemistry as a result of this tendency. Using QbD to create process analytical systems gives more regulatory versatility throughout the course of the method’s lifespan, which is an important feature. This is accomplished by putting more emphasis on the method’s overall performance objectives than on strict instrument requirements.

While there are limited publications on integrating Green Chemistry principles with QbD, existing studies demonstrate that combining these concepts in analytical procedures can enhance environmental sustainability while maintaining the accuracy of the specific High performance liquid chromatography methods.

One prominent trend in research on analytical QbD is the replacement of acetonitrile, a typical HPLC solvent, with ethanol as a means of integrating green ideas into the design process. Similarly, in UHPLC, the selection of ethanol is based on its availability from renewable sources and reduced harmful effects. However, the use of ethanol adds new crucial elements to the separation process, necessitating meticulous method parameter optimization during the formulation of the analytical procedure as well as the experimental design phases.⁹⁴

When considering the greenness of an analytical procedure, three key aspects should be taken into account: the specimen before treatment, the effectiveness of analytical tools, also the use of suitable waste treatment techniques in addition to disposal material reductions. Making an analytical technique more environmentally friendly usually starts with selecting the appropriate solvent, and there are established selection guides available to help with this decision. Parallel selection guidelines have been created for bases and acids. These guides are valuable for evaluating supplement compounds used in analytical methods. They focus on environmental, safety, and health concerns, particularly regarding the development of dangerous contaminants or difficulties with disposal. Y. Gaber used a comparable strategy.

Background of AQbD

Figure 2: The process of analytical method development in AQbD environment. Click here to view Figure

The main steps of the AQbD process are the analytical target profile (ATP), risk assessment and examination, effectiveness, durability, and development of the method operable design region (MODR), as the AQbD workflow illustrates. A brief explanation of the particular procedure and most popular techniques for that stage are included in each box.

The goals of an analytical method are set by an Analytical Target Profile, which also specifies the precise measurements to be taken and the requirements for success, including accuracy, precision, and range, that must be fulfilled.⁹⁵ Prior to beginning the method’s creation, it should be determined, outlining its goal but omitting to name the analytical approach. Therefore, the analytical procedure must be capable of accurately quantifying all identified or specified related substances and degradation products of an API, even in the presence of excipients and other drug product components. This should be achievable from the reporting limit up to 120% of their specification limit, maintaining an accuracy within ±20.0% of the actual value. An ATP supports regulatory bodies once the technique is approved, allowing for better ongoing method improvement. After defining the ATP, an analytical technique that meets the ATP criteria should be chosen. Various analytical techniques exist, each with its working principles, but HPLC is generally regarded as the most appropriate for impurity profiling analysis. It can be difficult to use due to the numerous operational factors, but if properly adjusted for the intended usage, it can produce results that are accurate and precise.

Response Modelling and Response Surface Design

A Response Surface Design is a more sophisticated classification of the DoE. In order to optimize a method’s operational effectiveness, this stage of the optimization process identifies crucial elements and how they interact with the prior screening design. A polynomial equation of higher order or a quadratic form can be used to describe the elements’ correlation with the responses after they have been studied at multiple scales (y = b0 + b1x1 + b2x2 + b11x1 + b22x2 + b12x1x2 + residual). For example, adding quadratic components to the model allows for the identification of nonlinear correlations between factors and responses, hence enabling the model to have curvature.⁵³ When factors are studied at more than two levels, the testing area has a repeated centre point to determine the mistake caused by the experiment.⁹⁶

Figure 3: a) A three-factor, two-level full factorial design consists of eight experiments, each represented by a green circle. Click here to view Figure

 b) A three-factor, two-level fractional factorial design, derived from the full factorial design, includes four experiments, each marked with a blue circle. c) A three-factor, three-level central composite design includes 15 experiments, which incorporates one experiment at the center point.

Response-surface designs include various experimental layouts, such as the Central Composite Design, Box-Behnken Design, Doehlert Design, and three-level full factorial design. Figure 5 illustrates the distribution of experiments in a three-dimensional space based on a Central Composite Design. When factor levels are restricted, a D-optimal design may be used to cover an asymmetrical experimental domain. Compared to screening designs, these designs necessitate significantly more experiments for an equivalent number of parameters. They are used to determine the mix of elements that predicts the best response since they can forecast responses for specific combinations of factors.

LC Method Development

This review includes a detailed examination of research articles published between 2016 and 2018, focusing on the application of Analytical Quality by Design (QbD) in the development of HPLC detection methods, particularly those involving UV measurements. This topic forms the second part of the review. The HPLC technique’s adaptability allows it to be applied to a variety of scenarios with various elution and retention strategies. This review covers instances where AQbD was used to create LC procedures for figuring out an assay or contaminants in tiny artificially created molecules. The representative sample matrix was one of three drug products: API, single dose, or a combination dosage. Furthermore, examples of utilizing different computer programs for DoE, examining obtained research results, and developing design space based on both.

Development of HILIC Method

Hydrophilic Interaction Liquid Chromatography (HILIC) is a less commonly used technique compared to reversed-phase HPLC. In HILIC, the mobile phase is polar, containing at least 3% aqueous phase along with an organic solvent that is water-soluble. The moisture from the mobile phase adheres to the silicon particles in the stationary phase, creating a water layer that imparts polarity to the stationary phase. Therefore, HILIC ought to be able to retain strongly polar molecules adequately. Moreover, the HILIC separation process may utilize a number of retention mechanisms. Therefore, creation of particular HILIC approach could result in a less reliable method. Since retained pattern during HILIC separations is often poorly understood, an organized AQbD strategy for HILIC technique development should be adopted. ⁹⁷

Combination of Drug Product

These days, combination medication products that contain more than one APIs are becoming more and more beneficial. Multiple targets in one dose form undoubtedly has benefits, and also it allows for considerably simpler drug product management for the patient throughout long-term treatment. The more APIs are used in a formulation, the more complex it becomes because each API exhibits unique chemical and physical characteristics. Therefore, if it is feasible to do so, the creation of only one single analytical technique for the characterization of a combination of therapeutic substances poses a significant challenge to all analytical scientists.

Optimization of Pharmacopeial Methods

Monographs from international pharmacopoeia are typically included in a literature evaluation at the outset of method development. In addition to saving time and money that would have been used on method development, adopting a pharmacopeial technique also ensures that the method will function reliably and fulfil its intended purpose. However, many pharmacopeial techniques that date back ten years or more may be considered archaic in modern times. Improved separations and up to ten times shorter analysis run times can be obtained using novel stationary phases containing sub-2 μm particles and advanced analytical instruments. The use of systematic techniques employing DoE for additional IP method optimization is encouraged by the likelihood that the method’s resilience was already evaluated by OFAT tests.

Impurity Profiling

 A medicinal substance’s impurity profile is defined as “a description of the identified and unidentified impurities present in a new medicinal substance” by the International Conference on Harmonization (ICH). The method of analysing data to determine each impurity’s biological safety is known as impurity profiling. The purpose of impurity profiling is to keep the API stable or functional. The foundation of the pharmaceutical industry is the mass production of medications since it provides unique, high-quality active ingredients. Purity is a dynamic concept that is closely related to developments in analytical chemistry. Impurities in APIs are becoming more and more popular. Lately, both the purity profile and the impurity profile have gained importance due to different regulatory requirements. Impurities are defined as compounds of organic origin other than pharmaceuticals, synthetic components, or undesirable chemicals that are still present in the API in the pharmaceutical industry. Impurity profiles are becoming significant because of different regulatory mandates. Organic materials or undesirable compounds that are still present in an active pharmaceutical ingredient (API) are referred to as impurities in the pharmaceutical business. Both the formulation process and the aging of the API can produce impurities. The identified and unknown impurities found in a typical batch of APIs made under specific, regulated manufacturing conditions are described in the impurity profile. One of the most crucial fields of study for the examination of contemporary industrial medications is this one. Guidelines on contaminants in new medications, goods, and residual solvents have been released by the International Conference on Harmonization (ICH). The effectiveness and safety of medications can be impacted by contaminants, even in trace amounts. Nowadays, synthetic medications are the most often used drugs, from which different final formulations are created. These formulations ensure bioavailability and therapeutic action by delivering medication in a stable, safe, and palatable form.

Other Methods

The value and effectiveness of software-assisted analytical techniques are demonstrated through their ability to optimize processes. Zoldhegyi et al.⁹⁸ used software modelling to develop an automated UHPLC method for separating ten pharmaceutical compounds. The goal was to create a robust technique that achieved baseline separation and faster scanning times. Initial tests indicated the use of gradient elution in reversed-phase chromatography with a C-18 column. The gradient was set between 10% and 95% B, and several factors were optimized through the Design of Experiments (DoE), including gradient duration (5–15 minutes), column temperature (30–60 °C), and mobile phase B (a 50:50 mixture of ACN and CH3OH). This resulted in twelve experimental runs. After analyzing the data in Dry Lab 4, a model that closely matched the experimental results was developed. A 3D resolution map was created to visually represent the model, highlighting the MODR (Region of Maximum Desirability), where the Critical Method Attributes (CMA) were successfully met. At the chosen maximum working point, the shortest analysis time and an acceptable critical resolution may have been achieved. An examination of robustness was performed for the selected working point.

Greenness Assessment for Development Method

The method integrates tri-combinations for analyzing two medications, with all three factors being essential in the development of procedures. A solution cannot claim to be environmentally friendly without undergoing proper evaluation using the appropriate methodologies. Four assessment tools were utilized to evaluate the environmental impact of the method: the AGREE-Analytical Greenness Metric, the AGMS-Analytical Method Greenness Score, the GAPI-Green Analytical Procedure Index, and the NEMI-National Environmental Methods Index. Each of these tools offers a range of features, limitations, and evaluation techniques. The results of each assessment instrument will determine which design is the most ecologically friendly as well as the evaluation method to employ. Although this approach evaluation procedure used multiple technologies, all of the data were presented in an ecologically responsible manner. This is how the methodology was assessed: the entire amount must not be more than 50 ml. Because there is little loss as a result of the recycling process, the third quadrant was given a green tint. The primary NEMI picture of a method.⁹⁹

NEMI

Nemi is a popular qualitative assessment method used to evaluate green chemistry. shown in (Figure 4). It was the only instrument available for assessing GAC efforts at first. NEMI offers advantages when examining the green analytical approach, even though it is creating new instruments for the GAC assessment. A sphere-shaped symbol with four quadrants and matching hues (green and colourless) is used to symbolize NEMI. In quadrant one, the EPA is in charge of handling the Persistent Bio-accumulative Toxic substances listed on the Toxicity Regulatory Inventory list.

In contrast, the Toxicity Compounds Regulation Inventory list of PBT compounds are subject of quadrant two’s work. Since the materials employed in this procedure are not included in PBT, this quadrant has a green hue. The second quadrant of the RCRA is occupied by hazardous substances, which are primarily regulated.

Because of the compounds that this method also discovered to be on the RCRA list, the second quadrant is displayed as green. The analytical solutions pH levels must be below a particular value, and the mobile phase ethanol plus phosphate buffer pH levels must be 60:40%. The 3rd region satisfies these criteria, making it a green zone. Waste is the topic of the fourth quadrant. The method showed perfect resilience, 100%, with the robustness module in Dry Lab 4 simulated a total of 729 experiments. Subsequently, the most significant parameter combinations were verified.

GAPI

GAPI, a somewhat altered NEMI, has 11 classifications and uses red, yellow, and green to denote hazard, tolerance, and environmental friendliness. In research work, ⁸⁴procedure make the use of GAO for evaluation easy through the development of freely available software. The application, in which it has 11 simple steps to get the result, must have the method details that need to be evaluated entered.

AMGS

An alternative method for assessing health, security, and the environment is provided by the HPLC Environmental Assessment-HPLC-EAT and SHE-Safety, Health, and Environmental Assessment, which are integrated with AMGS. The AMGS was divided into three categories: environmental health and safety, solvent energy, and equipment. These three scores add up to the method’s overall score, which should be as low as feasible to take into account making the method as environmentally friendly as feasible. The ultimate outcome obtained for the suggested methodology was 1242.90. in (Figure 4). which, when entering the data needed into the calculator accessible exclusively through AES Green Computing University illustrated the designed model’s favourable environmental impact.

Figure 4: [A] NEMI, [B] GAPI, [C] AMGS. Click here to view Figure

Agree with Metrics

AGREE WITH METRICS, the new Green Assessment instrument, contains all twelve of the green analytical concepts. The importance of the unique principles scoring obtained from the authorities of every individual was emphasized. Which was the overall result, which was represented as 1. The emphasis has placed on the individual principles score derived from each person’s rights. A value that is closer to one indicates how green the procedure is. After entering the specifics of the method into the application, (Figure 5). shows the overall result. Environmental effects of the approach have been characterized as “extremely benign” also “long-term sustainable.” While analysing approach’s greenness is done through five different methods or processes, the main goal was to determine how sustainable the method was. Whatever their strategies, every methodology revealed that this approach is safe for the environment and adaptable to future green assessments with no issues.¹⁰⁰

Figure 5: The AGREE metric provides the results of the proposed green assessment method. Click here to view Figure

Conclusion and Future Prospective

Pharmaceutical development and manufacture fundamentally rely on reliable analytical techniques, with High-Performance Liquid Chromatography (HPLC) and Ultra-Performance Liquid Chromatography (UPLC) being the most commonly utilised for drug product analysis. With the escalation of formulation complexity, the development of analytical methods necessitates enhanced precision, accuracy, and robustness. The Analytical Quality by Design (AQbD) framework provides a systematic, science-driven methodology that enhances method development by fostering a comprehensive understanding of analytical processes and establishing pre-defined objectives, such as the Analytical Target Profile (ATP).

Quality by Design (QbD) improves method reliability, decreases experimental workload, lowers costs and time, and guarantees optimal performance and reproducibility. The integration of AQbD with Green Analytical Chemistry (GAC) principles enhances the creation of ecologically sustainable techniques by minimising the use of hazardous solvents and reagents while preserving analytical efficiency. To evaluate environmental effect, several green assessment methodologies are employed, including AGREE (Analytical GREEnness Metric), AGMS (Analytical Method Greenness Score), GAPI (Green Analytical Procedure Index), NEMI (National Environmental Methods Index), and the Analytical Eco-Scale. These instruments offer quantitative and visual evaluations that inform the selection and enhancement of more sustainable analytical methods.

Implementing green methodologies outside the AQbD framework may jeopardise performance and necessitate revalidation. Integrating AQbD, GAC, and HPLC technologies improves process robustness, sustainability, and regulatory compliance. This integrated approach guarantees uniform analytical excellence while advancing enduring environmental objectives. The comprehensive implementation of these approaches signifies an advanced approach in pharmaceutical analytics, harmonising with quality assurance and environmentally sustainable procedures.

Acknowledgement

The authors would like to thank the University of South Africa for providing research infrastructure.

Funding Sources

The author(s) received no financial support for the research, authorship, and/or publication of this article.

Conflict of Interest

The authors do not have any conflict of interest.

Data Availability Statement

This statement does not apply to this article.

Ethics Statement

This research did not involve human participants, animal subjects, or any material that requires ethical approval.

Informed Consent Statement

This study did not involve human participants, and therefore, informed consent was not required.

Clinical Trial Registration

This research does not involve any clinical trials.

Permission to reproduce material from other sources

Not Applicable

Author Contributions 

Akanksha Mansing Valvi: Investigation, Formal analysis, Writing, review & editing.

Shantanu Sanjay Ghodke: Writing – review & editing, Formal analysis.

Rakesh Uttamrao Shelke: Writing – original draft, Writing – review & editing, Investigation, Conceptualization.

Dinesh Dattatray Rishipathak: Writing – review & editing, Supervision, Investigation, Formal analysis.

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Abbreviation

QbD ,Quality-by-Design; HPLC, High Performance Liquid Chromatography; GAC, Green Analytical Chemistry; PAT, Process Analytical Technology; GC, gas chromatography ;CE, capillary electrophoresis;CO₂,Carbon Dioxide ;LOV , Lab-on-a-Valve; NIR, Near-Infrared;  LC, Liquid Chromatography; IR, Infrared Radiation; GAC, Green Analytical  Chemistry; US GDP, Gross Domestic Product; AE, Atom Economy; PC, polycarbonate; COCl₂, dichloride ; CH3Cl₂,dichloromethane; C₂H₆O₂, ethylene glycol ;C₂H₄O, ethylene oxide; C₁₅H₁₆O₂, bisphenol-A;ICH, International Council for Harmonisation ;UHPLC ,Ultra-high performance liquid chromatography; ATP, Analytical Target Profile; AQbD , Analytical Quality-by-Design; MODR, Method operable Design Region; API, Active Pharmaceutical Ingredient; CMAs, Critical Material Attributes ;T, USP tailing; FMEA, Failure mode and effect analysis; DoE, Design of Experiments; HILC, Hydrophilic Interaction Liquid Chromatography ;GAO, Government Accountability Office ; OFAT, One-factor-at-a-time; AGREE, Analytical Greenness Metric, AGMS; Analytical Method Greenness Score; NEMI, National Environmental Methods Index; GAPI , Green Analytical Procedure Index; EPA, Environmental Protection Agency ;PBD, Polybutylene terephthalate ;RCRA ,Resource Conservation and Recovery ;HPLC-EAT, High Performance Liquid Chromatography (HPLC) Environmental Assessment Tool;

To Mesoporous Silica Nanoparticles

(MSN) was first introduced by Kuroda et al¹.Japanese researchers and Mobil Oil Company researchers in the United States began investigating it as early as 1990. Within the 21st century, nanotechnology has developed crucial innovations in medicate conveyance. Cutting edge nanotechnology has been contemplate a collective stage for investigate in affiliation with the advancement of cutting-edge technologies. Nanomaterial has been utilized altogether within the community care division since of its highlight to grasp, convey, ensure and provide restorative specialists, especially to the focused on tissue and gives security by decreasing measurements estimate and recurrence of administrations.²

In recent years, mesoporous silica nanoparticles had made development in medical and diagnostic applications due to their characteristics like as uniformity, tunable porosity, mesoporous properties, compatibility, easy operationalization and noncontact nature, as well as multifunctional drug delivery systems^3–5. Large mesoporous materials are constructed using self-build surfactant particles as a framework to condense silica prototypes in the surrounding area. Remove the framework and then construct the object in the mesh space. This new product family is characterized by the allocation of pores within the size range of 2 and 20 nm, a large pore size (about 1 cm³ g-1), an enlarged surface area (about 1000 m² g-1) and their location. The silicon-dense group will be beneficial for the next line of work. These properties make mesoporous silica nanoparticles an ideal candidate for applications that may require adsorbed materials, such as drug delivery systems, as first proposed by the Valletâ Regà group in 2001.⁶ In addition to having chemical properties similar to bioactive glasses, bioceramics with a mesoporous structure show bioactive behaviour. Characteristics of mesoporous silica nanoparticles are shown in table 1.^7,8

A huge wide variety of research has centered on the collaboration in-between surfactants and silica groups so that you can form a particular mesoporous silica nanoparticle. It’s miles terminated that mesostructural surfactant–silica nanocompoundes volutarily gather through interconnection of the natural and inorganic additives. Further to the thermostatics of the surfactant–silica assembly, the morphologies and dimensions of the ensuing compunds are mainly depending on the kinetics of sol–gel synthesis (including the heat of reaction, material water content, and pH value of the response answer). With a cautious manage of the self-arrangement and silica condensation charge, it’s miles feasible to tailor the sizes, mesostructures and arrangement of the mesoporous silica nanoparticles. Mesoporous silica nanocompounds, whose aspects include identical mesopores, clean modification and large compatibility, have received an awful lot of current interest for or their biomedical and catalytic applications.^9,10

Researchers have effectively studied on the implementation of these carriers for stacking an assortment of cargo extending from drug components to macromolecules such as proteins ^11,12 DNA ^13,14 and RNA. ^15,16 An inclusive set of written works are convenient, and investigations is still underway in analyzing unused roads for the utilization of MSNs in medicate conveyance. A few audits relating to MSNs in moving forward the solvency of the sedate, ^17,18 as controlled/sustained medicate conveyance structure¹⁹ applications in biomedicine have been distributed.^20,21

The mesoporous silica nano-compounds have one of the ideal characteristics, particularly in the large-scale stacking of useful operators and the ensuing discharges. Owing to solid Si-O bond, silica-based mesoporous nanoparticles are more steady to outside reaction such as debasement and mechanical push in comparision to niosomes, liposomes, and dendrimers which restrain the require of any outside stabilization within the blend of mesoporous silica nanoparticles.^22,23 The surface functionalization is for the most part required to stack appropriate sort of sedate particles (hydrophobic/hydrophilic or positive/negative charged), By modifying some activities using chemical connections with other materials, such as stimuli-responsive, luminous, or capping materials, they may also have normal standards or attributes, leading to clever and multifunctional features.¹⁰

Table 1: Characteristics of Mesoporous silica nanoparticles essential for drug delivery

Importance of topical drug delivery system

There is currently an increasing need for treatments that can improve patient compliance, so cosmetic or pharmaceutical delivery is considered whenever possible.³¹ The ability to deliver bioactive molecules through the skin represents an area of interest as an substitute to the peroral or parenteral injection. This transdermal infant bypass the intestinal tract, avoiding the pre-systematic metabolism, is pain-free and allows self-management. Local delivery has the potential to eliminate the need for referral and reduce the total amount of drug required, thus minimizing adverse effects on the target.³² The plant can be used to treat skin inflammation, photoaging, microbial and fungal infections and cancer.³³

Structural features of mesoporous silica nanoparticles

Mesoporous silica particle creation may have begun in the 1970s. The first business to create MSNs from aluminosilicate gels utilizing liquid lead liquid (Mobile Crystalline Materials or Mobile Composition of Matter) MCM-41 was Mobile Research and Development Corporation in 1992. According to IUPAC, mesoporous silica nanoparticles are substances with pore sizes, ranging from 2 to 50 nm and whose structural order determines the arrangement of their pores,^34–36 MCM41 has a hexagonal crystal system with a pore size of 2.5 to 6 nm and uses a cationic surfactant as a template. MCM41 is one of the most widely distributed drug products. Various mesoporous materials were also formed by changing the initial component and reactants. They can differ in their configurations or pore sizes. While MCM48 has a cubic structure, MCM-50 has a layered structure. Types of Mesoporous silica nanoparticles are shown in table 2 and fig 1.³⁷

Alkyl polyethylene oxide (PEO) oligomeric surfactants and polyalkylene oxides are examples of non-ionic triblock copolymers. Additionally, block copolymers have been used as models. The triblock polymer employed and the mesopore symmetry SBA-11 (cubic), SBA-12 (3D hexagonal), SBA-15 (hexagonal), and SBA16 (cubic lattice structure) have been chosen. To achieve the desired symmetry in the mesoporous material, the ratio of ethylene oxide to propylene oxide was altered. SBA-15’s porous structure is often used in biological applications ³⁸ Many other MSNs are the Technical University of Delft (TUD1), the Hiroshima Mesoporous Materials33 (HMM33), Research Centre for Chemistry and Catalysis (COK-12) and their pores differ in symmetry and shape.^39,40

Figure 1: Types of mesoporous silica nanoparticles   Click here to view Figure

Table 2: Various types of MSNs.

Synthesis of MSNs

Sol-gel technique

Changed Another term for the sol-gel method is Stober’s procedure. In a controlled catalytic environment, this process involves the hydrolysis and condensation of tetraethyl orthosilicate, the precursor to silica. This approach uses both basic and acid catalysts. The alkoxide group hydrolysis is determined by the reaction circumstances and the molar ratio of silica to water molecules. pH speeds up the rate of hydrolysis. This process is aided by condensation. Interlinking results from hydrolysis and condensation. Multiple conjugations that enter the gel’s crosslinking structure result in a chain structure. In order to get components with the desired size and enhanced properties, the sol-gel process is used.²⁵

Evaporation-induced self-assembly (EISA) method

Mahoney et al had first introduced the evaporation-induced self-assembly method. ²⁷ Another process for formulation of mesoporous silica nanoparticles and patterned thin films. This process involves a number of steps, including the formation of primary nanoparticles, the diffusion of structuring agents, such as ionic or non-ionic surfactants, and inorganic pioneers in ethanol and water; quick solvent evaporation to accomplish inorganic encapsulation and film formation; the balance between the film’s water content and the ambient air; the development and stability of hybrid mesophases; and, lastly, conjugation to solidify the network structure. Many structure-directing compounds facilitate the growth of ordered mesoporous metal oxides for photovoltaics and sensing applications.²⁸ The most popular forming agents are pluronic surfactants and amphiphilic block polymers. One of the main advantages of the EISA approach over another hydrothermal procedure is the quick synthesis of mesoporous silica nanoparticles.²⁹

Microwave Assisted Synthesis

It aids in the creation of less expensive mesoporous silica nanoparticles by forming them hydrothermally, where heating forces the nucleation process. As opposed to the traditional convection heating method, it reduces the time needed for synthesis and particle size and speeds up polymerization; the rate of swelling of the material is higher than that of material made by conventional heating. This technique is primarily used in scientific research. Higher heating to the crystallization temperature, more supersaturation because precipitated gels dissolve more quickly, faster crystallization time, and higher heating are only a few of its numerous benefits over other general approaches,³⁰ continuous nucleation is a result of ongoing heating during crystal development. Using this method, Wu et al. created a thermally stable hexagonal molecular sieve of MCM-41 molecules.⁵³

Ultra-sonic Synthesis

Run et al.⁵⁴ in 2004 discovered the ultrasonic synthesis technique for formation of mesoporous silica nanoparticles. This method results in the formation of well-organized hexagonal mesostructured with a large surface area of about 1100 m²·g⁻¹, the main pore size 22-30 angstrom and the volume of pore is about 1 cm³·g⁻¹. The one main merit of this synthesis technique is time required is highly reduce to minutes.⁵⁵

Mechanism

Both hydrolyzed silica adsorbs around the micelles, according to the data, and in the case of SBA-15, the silica and surfactant react in the preliminary phase to produce a core-shell, model ⁵⁶. They can forecast the modifications that will take place concurrently with the design process by using this model. It has been discovered that silicate ions have a tendency to draw in the vicinity of the surfactant micelles during the development phase when tetramethyl orthosilicate (TMOS) hydrolyzes first on the silicate surface (approximately 40 seconds). Tiny silica clusters occur as a result of the early hydrolysis and condensation of the silica pioneers, which also reduces the quantity of surfactant surrounding the micelles and their repulsive force. Transmission electron microscopy (TEM) examinations verified that the reaction mixture had sufficiently distinct hexagonally organized silica mesopores after around 400 seconds. This is in line with the “current knob model” of mesoporous silica nanoparticles production processes that was previously put out,^57,58 Small Accelerator Xray Scattering (SAXS) machine. This process works well when tetraethylorthosilicate (TEOS) is used alone as lead without other solvents such as ethanol. TEOS is an oily monomer that exhibits phase separation under static conditions, while an emulsion-like structure is attained under pressure. Primarily, cetyltrimethylammonium bromide (CTAB) forms ellipsoidal micelles with a core having hydrophobic tails. When TEOS is added, it dissolves in the hydrophobic core, thus expanding the micelle and causing the micelle shape to change from elliptical to spherical. When TEOS is hydrolyzed, the monomers become hydrophilic and are distributed into the liquid medium. The negatively charged TEOS hydrolyzable monomers are adsorbed onto the positively charged CTAB micelles via electrostatic attraction. When TEOS in the hydrophobic core is completely depleted, the micelles become minor and minor. Since the hydrolysis and condensation processes occur concurrently, the nanoparticles continue to form until all TEOS is hydrolyzed and a silica shell forms around the nanoparticles. Mechanism of formation of nanoparticles is shown in fig. 2. Adjacent nanoparticles clusters, leading to the growth of mesoporous structures.⁵⁹

Figure 2: Mechanism of formation of Mesoporous silica Nanoparticles   Click here to view Figure

Functionalization of MSNs

Modification of reaction parameters (relative amounts of alkoxysilane, water, catalyst) and temperature, the size, pore size, and morphology of mesoporous silica nanoparticles may be adjusted as desired.

Mesostructured Ordering

The pores available in MSN will possess different diameters based on which category of surfactant is used. The longer the length of the chain of the surfactant, the larger the pores of mesoporous silica nanoparticles, and the smaller the chain length, the smaller the pores of mesoporous silica nanoparticles ^6,54,60,61. TEOS concentration affects the mesostructure arrangement of the goods. Greater amounts of TEOS show weak mesoporous structures, while smaller amounts are not sufficient to form mesoporous structures. ⁶² The amount of the surfactant CTAB was also found to have a significant effect on the microstructural properties of the material. A less surfactant concentration cannot form micelles, so the resulting nanoparticles will not have a structure, while excessive CTAB concentration can cause damage. ¹⁴ The addition of N,Ndimethylhexadecylamine (DMHA) can act as a pore modifier, thus helping to maintain the required pore size.⁶³ Size of pore have direct impact on drug release rate which seen in ibuprofen.^64,65

The mesostructured organization and pore size of the noncompounds are significantly impacted by the kind of surfactant used. Based on the alterations in the counterions shown in cetyltrimethylammonium (CTAB), the effect of templating agents was examined. Mesoporous, interdimensional silica nanoparticles were made using cetyltrimethylammonium chloride (CTAC) as a pore-forming template. The pore radius grew and the pore architecture changed from cylindrical to radiant after altering the reaction to a bigger tosylate ion (CTATOS).⁶⁶ In order to address the drawbacks of conventional mesoporous silica nanoparticles, such as their tiny pore size and low particle size, researchers have recently used novel ways to alter the performance of MSNs. Given this, Huang et al.⁶⁷ combined semi-fluorinated short chain anionic fluorocarbon surfactants, Capstone FS66 and CTAB, to create extremely monodisperse diphosphates with wide pores and dendritic morphology utilizing a unique dual templated solgel reaction nanoparticles. They carried out a qualitative investigation that demonstrated morphological alterations brought about by the addition of Capstone FS66. The particle has a dendritic channel pore structure and is larger. The picture transforms into a big dendritic structure that resembles a flower when more material is added. Yu et al. used the same concept to create dendritic MSNs with particle sizes greater than 200 nm by using Pluronic F127 as a particle growth inhibitor and imidazolium ionic liquids with varying alkyl lengths as cosurfactants. They discovered that the size of mesoporous silica nanoparticles was unaffected by either time or temperature.⁶⁸

Control of Shape

Mesoporous silica nanoparticles’ cellular absorption and biodistribution are significantly influenced by their form. Thus, it is essential to tightly regulate the form of MSNs in order to regulate their excretion and other activities.⁶⁹ Article by Huang et al.⁷⁰ The potential of spherical MSNs for medication delivery has been extensively studied up to this point. The utilization of aspheric mesoporous silica nanoparticles is uncommon, nevertheless. Nonspherical materials in the form of cubes, flakes, films, sheets, ellipsoids, and rods will be created by carefully controlling the reaction. The shape of MSNs was discovered to be influenced by the molar concentrations of TEOS, water, base catalyst, and surfactant. Cai et al. created MSNs with a variety of morphologies, including spheres, silicon rods, and micron-sized oblate silica, by varying the amounts of TEOS, NaOH/NH4OH, and CTAB ⁷¹. By adjusting the concentration of dodecanol as soft template and the synthesis temperature, various silica particles with structures ranging from circular to shell-line, football-shaped, peanut-shaped, hollow and yolk-shell-like structures can be made. Six variables were created that provide control over the size, porosity, internal void and shell structure. Their analysis showed that the addition of dodecanol produced soft samples that produced different products with different morphologies.⁷² This indicates that any alteration to the nanoparticle structure during the first phase will result in a change to the morphology of the particles.

With varied ratios, MSNs may take on a variety of forms. Spherical MSNs often produce rod-shaped MSNs. By altering the sol-gel reaction’s negative reactions, they may be obtained. Rod-shaped MSNs may be produced by varying the temperature, introducing cosolvents such heptane, increasing the catalyst concentration, and altering the molar content of the reactants. ^64,70 It is quite difficult for the cylinders to retain their shape due to the decrease in inertia force. These MSNs can be synthesized by adding surfactants⁷³ adding potassium chloride and ethanol. ⁷⁴ By adding a small amount of ammonium fluoride and heptane, ⁷⁵ a non-ionic block copolymer P104⁷⁶ is obtained using the cetyltrimethylammonium bromide-sodium dodecyl ternary surfactant system. Using a ternary surfactant system of cetyltrimethylammonium bromide–sodium dodecyl sulfate–Pluronic123.⁷⁷

Drug Loading

The adsorption properties of mesoporous silica nanoparticles primarily determine the drug carrying capacity. Mesoporous silica nanoparticles may include hydrophilic and hydrophobic bundles within their pores. Because of their greater volume, MSNs may handle loads greater than conventional supports. Nonetheless, a lot of study has been done on improved medication delivery. Enhancing mesoporous silica nanoparticles loading may be achieved by the synthesis of HMSNs. Octadecyltrimethoxysilane (OTMS), 3-aminopropyltriethoxysilane (APTES), and 3-cyanopropyltriethoxysilane (CPTES) are some of the silanes that He et al. attempt to functionalize. To improve the loading of 5-fluorouracil (5-FU) in hollow mesoporous silica nanoparticles, surface silanol groups are used. Compared to 18.34% of free hollow mesoporous silica nanoparticles, the loading capacity of amine-functionalized HMSN was improved by 28.89%. Amino modified hollow mesoporous silica nanoparticles and negatively charged 5-FU may interact electrostatically to accomplish this. via altering the operating mode, a similar tactic to the chemical transport capacity via electrostatic attraction may be used.^78,79 Because of their vacant area, hollow mesoporous silica nanoparticles have been shown to be better carriers in terms of loading than MSNs. Compared to mesoporous silica nanoparticles, hollow mesoporous silica nanoparticles has a 3–15 times greater drug loading. Additionally, the same carrier was used to carry two medicines.⁸⁰ Using a polymer gate to trap hydrophobic materials may further increase the loading capacity of MSNs.⁸¹

Drug Release

Mesoporous silica nanoparticles’ drug release profile is mostly reliant on various pores, and by altering the MSNs’ surface, medications may be tailored to suit biological requirements. The cooperation between the drug molecules and the pore’s surface groups determines how the release is controlled.⁸² It was found that activating the drug first and then loading it to the active site with the amine group played a crucial role in promoting the release of the components compared to the systems where the drug was first activated and then loaded. This may be due to loading the drug into the pores and closing the APTES against drug discharge. If the site is first activated and then loaded, the drug will be adsorbed on the mesoporous silica nanoparticles surface, which will cause an explosion. investigated the role of APTES concentration on drug release and the results showed that changes in APTES concentration played an vital role in regulating drug discharge from the pores.⁸³

Applications of Mesoporous silica nanoparticles in tropical drug delivery system

MSNs for Anti-Inflammatory applications

Different antibiotics have been hosted on MSNs and their release rates have been regulated thanks to their unique properties. There are two silica solutions: MCM-41 and SBA 15. Ibuprofen was carried by a variety of channels, and the response of its drug discharge was examined. Three phases of drug discharge were identified during the analysis of the data: first, the drug adsorbed on the MSN surface releases quickly, followed by a sustained release of the drug components in the surface’s tiny pores. Particles in MSNs have varying sample orientations. When the medication is fully integrated into the big pores of the particles, the last step of delayed discharge takes place.⁸⁴ Most of the studies on the effective delivery of anti-inflammatory drugs by MSNs have used ibuprofen as an ideal drug. Works have been done to modify drug discharge by numerous changes of the silanol group. Surface functionalization has been shown to alter and enhance drug discharge.^64,85

Mesoporous Silica Nanoparticles for Antitumour therapy.

In an effort to enhance site-specific medication delivery and avoid negative responses, more research is being done on the surface functional characteristics of MSNs. It is a multipurpose vehicle that can be customized to provide effective therapy and loaded with medications that have unique physicochemical properties. Because of the effects of enhanced permeability and retention (EPR), they may group together in tumors.⁸⁶  Increased cellular uptake of active medications is facilitated by active targeting. The right receptors found on tumor cells are selected, and particular ligands for these receptors are affixed to the surface of MSNs in accordance with the differences between normal and tumor cells. Target ligands may be loaded onto MSNs to provide targeted medication delivery. One known ligand that enhances the function of folate receptors on tumor cells is folate (FA).

The carboxyl group of FA and the amine group of aminopropyltriethoxysilane (APTES) form an amide bond, which facilitates FA incorporation on the surface of MSNs. In order to fight B16F 10 skin cancer, Ma et al.⁸⁷ attached folic acid to the surface of HMSN and used 5-aminoethylpropionic acid for photodynamic treatment. It was shown that the development exhibited strong photosytotoxicity to tumor cells. According to the same study, amine functionalization on the surface makes it easier for folate to bind covalently to the receptor site so that tumor cells may preferentially absorb doxorubicin (DOX). Studies on cellular intake and apoptosis revealed that FA-MSN-NH2-DOX had a higher ability for cellular absorption than MSN-NH2-DOX.⁵⁵

Gated Drug Release/Controlled Drug Delivery

Studies will continue to get intelligent, controlled release of the pharmaceuticals by capping the floor of the pores, even if the duration and the pore structure changed the drug launch charge. The gated launch makes it possible to transmit medications in a modified and intelligent manner to the target web page using MSNs. Only in response to certain stimuli, like as pH, temperature, enzymes, redox, and so on, do the gates of the pores open. When it comes to preventing dangerous adverse medication reactions to other organs, the gated drug launch principle may be quite effective.⁸⁷ Several reviews on this consideration have been posted, some of which we’ve mentioned within the following segment.

pH-Responsive Drug Release

pH is a general science that determines drug release due to the wide range of pH values in the human body. Prednisolone loaded MCM48 particles were coated with succinate β-poly lysine (SPL) to provide pH-sensitive discharge of the drug in the colonic region. In vitro release procedure presented sustained drug release, indicating the success of the pH-sensitive drug delivery approach. At acidic stomach pH, SPL blocks drug discharge due to its ionized form, while at colonic pH, SPL converts to an ionized form that promotes molecular diffusion through MSNs. The produced nanoparticles can be used as an supportive therapy for intestinal diseases (colon cancer and colorectal cancer).⁸⁸

Redox Responsive Drug Release

An other tactic often used to control carrier emissions is redox triggering. This gadget delivers antibodies using endogenous antibiotics. For this drug delivery, redox cleavable disulfide bonds are often used. For redox-sensitive drug delivery, Wang et al. created a disulfide-linked polyethylene glycol (PEG) that is affixed to mesoporous silica nanoparticles. Using rhodamine B (RhB) as a model medication, an in vitro release study evaluated the performance of the prepared MSNs. The release media was supplemented with glutathione (GSH) in an amount equivalent to the intracellular concentration. In the absence of GSH, it was shown that RhB discharge was negligible, demonstrating the cap’s ability to stop drug release. Furthermore, PEG surface modification gives the nanoparticles a high degree of biocompatibility.⁸⁹

Temperature Responsive Drug Release

Thermosensitive mesoporous silica nanoparticles are also increasingly investigated as a way to control drug discharge. In this case, Bathfield et al. established copolymeric nanoparticles built on PEO-b-poly(N-isopropylacrylamide) as a model conducting material for the formulation of functionalized mesoporous silica nanoparticles. The sample drug, ibuprofen, was loaded into the mesopores using a well where the drug was directly integrated into the hybrid product. Finally, the structure-modifying agents in the development were drug-loaded polymer nanoparticles. Drug delivery profiles at 20°C and 45°C showed temperature-sensitive parameters drug release at 450 °C was higher than that at 200 °C.⁹⁰

Chemical and Enzyme Responsive Drug Release

Many drugs and enzymes found in the body or prepared in bacteria have also been investigated for their ability to increase drug release from MSNs. Researchers have investigated the feasibility of glucose-based drugs in terms of drug delivery and their effectiveness in diabetes management. ⁹¹In a particular study, mesoporous silica nanoparticles with the function of the signal reporter alizarin compound ketone (ALC) were created. Glucosylated insulin is then absorbed into the pore via benzene-1,4 diboric acid (BA) -mediated esterification. Together it as a hypoglycemic agent and a pore blocker. Furthermore, rosiglitazone maleate was absorbed into the pore, forming multifunctional MSNs. Competition between ALC and BA occurs in the presence of glucose, which causes the pores to open and the medicine to be released.⁹²

Current and Future perspectives

A few medicines have been granted by the FDA for therapy and use, but these new techniques have achieved significant breakthroughs in disease treatment and hold the potential to transform traditional approaches to treatment and diagnosis. Since the potential of employing MSNs as drug delivery vehicles was found, substantial study has been performed to determine the importance of this technology in the treatment of numerous illnesses. The majority of research have focused on the use of vectors for targeted anticancer medication delivery.

Although many studies confirm the effectiveness of mesoporous silica nanoparticles (MSNs) in topical delivery, some of the data may be explained differently. Certainly, the improvements in drug penetration and efficacy seen from MSNs can be caused by surfactants, penetration enhancers or co-administered components in the formulation, among other things. Likewise, how MSNs release their payload over time can depend not only on their pore structure and coatings, but also on pH, enzymes or the skin’s moisture level which change from one setting to another. Often, results seen in experiments with cell cultures do not match what happens in human skin. When we think about these options, we understand why we need thorough study and examination to clearly identify the influence of MSNs in therapeutic practices.

However, regulations and restrictions prevent the secure and effective interpretation and legal authorization of these compounds. In contrast to other nanocarriers, the production process of mesoporous silica nanoparticles is simple and efficient. In addition, these MSNs have additional potential due to their use as various nanocarriers for drug surfaces and bodies, as well as for theranostic and imaging purposes, as well as supporting various drugs. In this context, cell research and preclinical studies have achieved remarkable results. However, there are still many challenges in successfully translating this platform into hospitals. It may be very difficult to synthesize MSNs with the same properties and quality. The development of the technology mainly depends on the production capacity, therefore, the formulation of mesoporous silica nanoparticles at the manufacturing stage may have an impact on their commercialization. To ensure product recyclability, manufacturing processes need to be better understood and managed. Furthermore, since all medications are unlikely to be loaded uniformly, the value of mesoporous silica nanoparticles might vary depending on the context, which may aid in determining the maximum dosage of mesoporous silica nanoparticles. Chen et al. employed several analytical methods, such as customized fluorescence correlation spectroscopy (FCS) in conjunction with particle size/gel permeation chromatography (GPC), to assist preserve size and long-term stability, minimizing batch-to-batch variability. Although the mortality of most inorganic nanoparticles remains a significant worry, promising studies on the effectiveness and biocompatibility of mesoporous silica nanoparticles in animal models indicate that this platform might be adapted for medical usage.⁹³

Conclusion

Advanced drug delivery systems for the skin often use MSNs, thanks to their ability to be adjusted, a large surface area and many surface uses. A review of new research demonstrates that MSNs support controlled drug release, increase the stability of drugs and help achieve targeted therapy for dermatology. This is especially useful in treats skin problems that continue for a long time since it releases medicine slowly right where it’s needed. The data I reviewed highlights that MSNs are now more widely used for anti-inflammatory, anticancer and stimulus-controlled drug delivery. Still, bringing MSN-based systems from the lab to clinical practice is currently a challenge. The problems of repeatability, safety with prolonged use, mass production and okaying by regulators are not fully solved yet. Informed by recent experiments, the conclusions here suggest an encouraging, but evolving, setting for MSN-enabled treatments. More interdisciplinary work is needed to solve current issues and make MSNs suitable for use in dermatology.

Acknowledgement

I would like the express their sincere gratitude to :Pres’s College of Pharmacy (for Women), chincholi, nashik for providing the necessary resources and support. Savitribai Phule Pune University, for its academic framework and infrastructure.I also acknowledge the contributions of the reviewers and editors for their valuable feedback and suggestions.

Funding Sources

The author(s) received no financial support for the research, authorship, and/or publication of this article.

Conflict of Interest

The authors do not have any conflict of interest.

Data Availability Statement

This statement does not apply to this article.

Ethics Statement

This research did not involve human participants, animal subjects, or any material that requires ethical approval.

Informed Consent Statement

This study did not involve human participants, and therefore, informed consent was not required.

Clinical Trial Registration

This research does not involve any clinical trials.

Permission to reproduce material from other sources

Not Applicable

Author Contributions:

Gauravi kherde: Conceptualization, Methodology, Writing – Original draft

Khaire Rahul: Analysis, Writing- Review & Editing

Kunde Vikas: Visualization, Supervision

Katkade Pratiksha: Reviewing, Editing

Pooja Akoshkar : Reviewing, Data Collection

Komal Taru: Reviewing

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The Podostemaceae generally known as river-weeds are distinctive aquatic angiosperms found in wetlands throughout the tropics and subtropics worldwide.^1-3 These plants thrive in fast-flowing, turbulent currents, firmly adhering to rock surfaces during the rainy season. As water levels recede during the dry season, they germinate, blossom, produce fruit, and eventually wither. During the rainy season, seeds dispersed by running water attach to rock surfaces, where they germinate and grow into seedlings.⁴ The subfamily Tristichoideae of Podostemaceae consists of approximately 20 species distributed across five genera globally, with Asia ^5-7 being the centre of species diversity. The five genera in this subfamily are Terniopsis, Tristicha, Indodalzellia, Indotristicha, and Dalzellia.^8-7 Out of these genera, the distribution map of Dalzellia ⁶ shows that Dalzellia ubonensis, D. ranongensis, D. kailarsenii, and D. angustissima occur in Thailand; and D. zeylanica and D. gracilis, in India and Sri Lanka, and India, respectively.

During floristic surveys conducted in the Kallar River (Latitude 9.240924^o, Longitude 76.965226^o), a tributary of the Pampa River in the Vadasserikkara Range of the Ranni Forest Division, Pathanamthitta District, Kerala, India, the authors collected several intriguing specimens of Dalzellia.

Taxonomic evaluation of the specimens, along with a review of relevant literature and molecular studies, revealed that the species collected from the aforementioned area corresponds to Dalzellia ubonensis M. Kato. Dalzellia gracilis, reported from India, is a closely related species to Dalzellia ubonensis. However, it differs from D. ubonensis in several key characteristics: the presence of roots, numerous root-borne shoots, and a crustose shoot that is irregularly shaped and nearly ribbon-like with hapters. Additionally, D. gracilis lacks the cupula at the base of the floriferous shoot and does not have “rosettes” on the upper side of the crust,^6,11 which are present in D. ubonensis. Morphological studies, supported by literature citations and robust genomic analysis, confirm that this is the first report of Dalzellia ubonensis from the state of Kerala, India. The voucher specimens were deposited at Kerala University herbarium (KUBH11308) and at the Post Graduate and Research Department of Botany, NSS College, Pandalam (NSSPDMBODHOS7) respectively. A detailed account of the species is presented below.

Taxonomic Treatment

Dalzellia ubonensis M. Kato, Acta Phytotax. Geobot. 57(1): 10 (2006).

The shoot of Dalzellia ubonensis is crustose (foliose), slightly pinkish in color, and adheres to rocky substrates like a foliose lichen, with no roots present ^10-11 (Fig.1B). The shoot measures 3–10 mm or wider and is lobed. It exhibits dorsiventral construction, with subdimorphic leaves (Fig.1A) arranged on the upper surface. The lower surface of the shoot, which attaches to the substrate, is devoid of leaves (Fig.1A).

Dorsal leaves

Arranged in branched longitudinal rows (Fig.1G), these leaves are densely fimbriate, linear-oblong, and smaller than the marginal leaves, measuring approximately 1 mm × 0.1 mm. They are rounded at the apex and oriented toward the distal end of the row.

Marginal (lateral) leaves

Narrowly deltoid, fimbriate, and rounded at the apex, these leaves are larger, reaching about 2 mm × 0.5 mm.

Rosette leaves

Scattered on the dorsal surface of the shoot, these leaves are linear, rounded at the apex, and measure approximately 2 mm × 0.2 mm.

Flowers are scattered on the shoots

Peduncle: 5–9 mm long (Fig.1C,1H).

Calyx: Membranous tepals as long as the ovary, shallowly 3-lobed, with deeper incisions at anthesis.

Stamens: Three, with di-thecous anthers, each filament 2 mm long, and equal brown (Fig.1H) lobes longer than the ovary.

Ovary: Obovoid-ellipsoid, about 2 mm long and 1 mm thick, trilocular with axile placentation; ovules number roughly 30 per locule.

Stigmas: Three (Fig.1C), papillate, 0.2–0.3 mm long.

Capsule: Stalked (10 mm long), dark brown, trigonous, measuring approximately 2 mm × 1 mm, with nine ribs. After dehiscence, the valves curve inward (Fig.1D), revealing numerous seeds, about 30 per locule (Fig.1E).

Figure 1: A- Dorsal surface of thallus with dimorphic leaves (a-marginal leaves, b-dorsal leaves); B- Ventral surface; C- Flower with a- stigma, b-anther, c- perianth; D- Dehisced fruit with valve refluxed inward;Click here to view Figure

Phenology

We observed both flowers and fruits during January (in India).

Distribution: Thailand, Vietnam, Laos, India.

Phylogenetic Studies

DNA Extraction, Amplification and Sequencing

DNA Barcoding using universal primers of MATK

DNA isolation using NucleoSpin^® Plant II Kit (Macherey-Nagel)

About 100 mg of the tissue/mycelium is homogenized using liquid nitrogen and the powdered tissue is transferred to a microcentrifuge tube. Four hundred microlitres of buffer PL1 is added and vortexed for 1 minute. Ten microlitres of RNase A solution is added and inverted to mix. The homogenate is incubated at 65^oC for 10 minutes. The lysate is transferred to a Nucleospin filter and centrifuged at 11000 x g for 2 minutes. The flow through liquid is collected and the filter is discarded. Four hundred and fifty microlitres of buffer PC is added and mixed well. The solution is transferred to a Nucleospin Plant II column, centrifuged for 1 minute and the flow through liquid is discarded. Four hundred microlitre buffer PW1 is added to the column, centrifuged at 11000 x g for 1 minute and flow though liquid is discarded.  Then 700 µl PW2 is added, centrifuged at 11000 x g and flow through liquid is discarded. Finally, 200 µl of PW2 is added and centrifuged at 11000 x g for 2 minutes to dry the silica membrane. The column is transferred to a new 1.7 ml tube and 50 µl of buffer PE is added and incubated at 65^oC for 5 minutes. The column is then centrifuged at 11000 x g for 1 minute to elute the DNA. The eluted DNA was stored at 4^oC.

Agarose Gel Electrophoresis for DNA Quality check

The quality of the DNA isolated was checked using agarose gel electrophoresis. 1µl of 6X gel-loading buffer (0.25% bromophenol blue, 30% sucrose in TE buffer pH-8.0) was added to 5µl of DNA. The samples were loaded to 0.8% agarose gel prepared in 0.5X TBE (Tris-Borate-EDTA) buffer containing 0.5 µg/ml ethidium bromide. Electrophoresis was performed with 0.5X TBE as electrophoresis buffer at 75 V until bromophenol dye front has migrated to the bottom of the gel. The gels were visualized in a UV transilluminator (Genei) and the image was captured under UV light using Gel documentation system (Bio-Rad).

PCR Analysis

Table 1: PCR Analysis

Primers used

Table 2: Primers used

The PCR amplification was carried out in a PCR thermal cycler (GeneAmp PCR System 9700, Applied Biosystems).

PCR amplification profile

MATK

98 ^oC   –              30 sec

98 ^oC   –              5 sec

45 ^oC   –              10 sec           }10 cycles

72 ^oC   –              15 sec

98 ^oC   –              5 sec

50 ^oC   –              10 sec           }30 cycles

72 ^oC   –              15 sec

72 ^oC   –             60 sec

4 ^oC   –             ∞

Agarose Gel electrophoresis of PCR products

The PCR products were checked in 1.2% agarose gels prepared in 0.5X TBE buffer containing 0.5 µg/ml ethidium bromide. 1 µl of 6X loading dye was mixed with 4 µl of PCR products and was loaded and electrophoresis was performed at 75V power supply with 0.5X TBE as electrophoresis buffer for about 1-2 hours, until the bromophenol blue front had migrated to almost the bottom of the gel. The molecular standard used was 2-log DNA ladder (NEB). The gels were visualized in a UV transilluminator (Genei) and the image was captured under UV light using Gel documentation system (Bio-Rad).

ExoSAP-IT Treatment

ExoSAP-IT (GE Healthcare) consists of two hydrolytic enzymes, Exonuclease I and Shrimp Alkaline Phosphatase (SAP), in a specially formulated buffer for the removal of unwanted primers and dNTPs from a PCR product mixture with no interference in downstream applications.

Five micro litres of PCR product is mixed with 0.5µl of ExoSAP-IT and incubated at 37^oC for 15 minutes followed by enzyme inactivation at 85^oC for 5 minutes.

Sequencing using BigDye Terminator v3.1

Sequencing reaction was done in a PCR thermal cycler (GeneAmp PCR System 9700, Applied Biosystems) using the BigDye Terminator v3.1 Cycle sequencing Kit (Applied Biosystems, USA) following manufactures protocol.

The Sequencing PCR mix consisted of the following components:

SequencingPCR amplification profile       

96^oC    –           2min

96^oC    –           30sec

50^oC    –           40sec              } 30 cycles

60 ^oC   –           4min

4 ^oC     –           ∞

Post Sequencing PCR Clean up

Mix D/W, 125mM EDTA, 3M sodium acetate pH 4.6 and ethanol were prepared and were properly mixed.

50 µl of mix was added to each well in the sequencing plate containing sequencing PCR product.

Vortex by Mixmate vortex and incubated at room temperature for 30 minutes

Spun at 3700 rpm for 30 minutes

Decanted the supernatant and added 50 µl of 70% ethanol to each well

Spun at 3700 rpm for 20 minutes.

Decanted the supernatant and repeated 70% ethanol wash

Decanted the supernatant and air dried the pellet.

The cleaned-up air dried product was sequenced in ABI 3500 DNA Analyzer (Applied Biosystems).

Sequence Analysis

The sequence quality was checked using Sequence Scanner Software v1 (Applied Biosystems). Sequence alignment and required editing of the obtained sequences were carried out using Geneious Pro v5.1¹²

Phylogenetic Analyses

The obtained DNA sequences were analyzed using the NCBI Nucleotide BLAST (BLASTn) tool (https://blast.ncbi.nlm.nih.gov/) to compare them with existing sequences in the GenBank database for species identification and similarity assessment. The sequences were submitted in FASTA format, and MEGA-BLAST, an algorithm optimized for highly similar sequences, was used for analysis. The results were filtered based on 100% query coverage and ≥98% sequence identity to ensure accurate species identification. The BLAST output provided a ranked list of matching sequences, displaying key parameters such as Max Score, E-value, Identity Percentage, and Query Coverage. The top hits were carefully examined, and sequences with high similarity consistently matched species within the genus Dalzellia (Fig.4). The strong alignment between the obtained sequences and the reference sequences in the database confirmed the taxonomic identity of the plant sample. This analysis provided molecular validation of the plant’s classification, demonstrating a high degree of confidence in its genetic identification.

Figure 4: Phylogenetic tree obtained from BLASTClick here to view Figure

Result

From the phylogenetic analysis the plant specimen collected from Kallar river (Vadasserikkara range) of Pathanamthitta district of Kerala, India was identified as belonging to Dalzellia species. Further detailed studies on morphology using the protologue made it clear that the plant specimen is Dalzellia ubonensis M. Kato (2006:10).

Discussion

The present study marks the first confirmed report of Dalzellia ubonensis M. Kato from Kerala, India, thereby extending the known geographic range of the species beyond its previously documented distribution in Southeast Asia, including Thailand, Vietnam, and Laos.¹⁴ This finding is significant in the context of Podostemaceae biogeography, as it underscores the potential for overlooked or undocumented diversity within Indian aquatic ecosystems, particularly in the Western Ghats region, a recognized biodiversity hotspot.

The taxonomic identification of D. ubonensis from the Kallar River was substantiated through a combination of morphological traits and molecular evidence. Morphologically, the collected specimens exhibited a crustose to foliose shoot system, subdimorphic leaves, and floral characteristics that are congruent with the original protologue of D. ubonensis. ¹⁴ These features contrast with those of Dalzellia gracilis, the only other species of the genus previously reported from India.⁶ In particular, the presence of foliose shoots and the distinctive floral morphology—such as the shape and arrangement of tepals and stamens—clearly differentiate the Kerala specimens from D. gracilis, which is characterized by a more gracile habit and differing reproductive structures.

Comparative molecular analysis, using the MATK gene as a barcode marker, further reinforced species-level identification. The high sequence similarity (≥98% identity) and complete query coverage in BLASTn analyses against verified D. ubonensis sequences provide strong phylogenetic support for the identification. This integrative approach aligns with recent trends in aquatic plant taxonomy, where DNA barcoding has become a valuable tool in resolving taxonomic ambiguities, particularly in morphologically plastic or cryptic species.

Ecologically, the presence of D. ubonensis in the Kallar River is consistent with the habitat preferences reported in prior studies from Southeast Asia, where the species is known to inhabit submerged rock surfaces in clear, fast-flowing rivers. ¹⁴ The specificity of this microhabitat underscores the narrow ecological amplitude of Podostemaceae members and raises concerns regarding their vulnerability to hydrological alterations. Previous studies have documented that Podostemaceae species are highly sensitive to changes in flow regime, sedimentation, and water pollution, ¹ all of which are increasingly common in riverine systems of the Western Ghats due to anthropogenic pressures.

The discovery of D. ubonensis in India has important implications for the regional flora and prompts a re-evaluation of the genus Dalzellia within the Indian subcontinent. It suggests that historical under-sampling or taxonomic oversight may have led to an underestimation of species diversity within this genus. Given that Podostemaceae are often restricted in distribution and exhibit high levels of endemism, our findings call for a more comprehensive survey of aquatic habitats across the Western Ghats and northeastern regions of India, where similar ecological conditions may support undiscovered or unreported species.

Conclusion

In conclusion, this study contributes to the growing body of literature emphasizing the importance of integrative taxonomy in documenting plant diversity. By confirming the presence of Dalzellia ubonensis in Kerala through detailed morphological and molecular evidence, it fills a critical gap in the known range of the species and highlights the Kallar River ecosystem as a significant site for aquatic plant diversity. Future research should focus on population genetics, reproductive ecology, and habitat monitoring to support effective conservation of D. ubonensis and related taxa in India.

Acknowledgement

The authors are grateful to management of NSS College, Pandalam and to the Post Graduate and Research Department of Botany for all the facilities provided.

Funding Sources

The author(s) received no financial support for the research, authorship, and/or publication of this article.

Conflict of Interest

The authors do not have any conflict of interest.

Data Availability Statement

This statement does not apply to this article.

Ethics Statement

This research did not involve human participants, animal subjects, or any material that requires ethical approval.

Informed Consent Statement

This study did not involve human participants, and therefore, informed consent was not required.

Clinical Trial Registration 

This research does not involve any clinical trials.

Permission to reproduce material from other sources

Not Applicable 

Author Contributions

Fouzia Hilal: Conceptualization, Methodology, Writing – Original Draft, Data Collection, Analysis, Writing – Review & Editing.

Jithesh Krishnan Ramakrishnan Nair: Data collection, Visualization, Supervision, Project Administration.

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