Wound Healing Infection Research Articles

Metal–phenolic nanozyme based microneedle patch with antibacterial and antioxidant for infected wound healing

Chronic bacterial-infected wound healing is becoming increasingly severe, with high rates of mortality and disability, owing to bacterial film, excessive accumulation of reactive oxygen species (ROS), inflammatory, and traditional therapeutics with poor drug permeability. Herin, a functional Zn-gallic acid nanozyme (Zn-NM) with excellent antibacterial and antioxidant capacity was incorporated with a gelatin-based microneedle patch (Zn-NM@MN) to achieve transdermal and sustained release of the drug at the infected wound site. The Zn-NM displayed the concentration-dependent antibacterial capacity, and 250 μg/mL of Zn-NM simultaneously possessed excellent antibacterial (89.36 ± 0.95 % for Escherichia coli, 92.44 ± 11.03 % for Staphylococcus aureus, and 95.03 ± 1.06 % for Methicillin-resistant Staphylococcus aureus), antioxidant properties and negligible cytotoxicity. After that, Zn-NM@MN could transdermal the epidermis or biofilm to sustain the release of Zn-NM for 3 h (release of 80 % drug). Systematic tissue regeneration assessment on rats’ infected full-thickness skin wounds demonstrated an enhanced wound healing rate. Zn-NM@MN could efficiently kill the bacteria (about 85 %), alleviate oxidant stress, reduce bacterial-induced inflammation, and promote vascular regeneration. This synergetic therapy strategy will pave the way for treating complicated infection wounds.

Polyvalent copper oxide nanozymes: Co-delivery of ROS and NO for inducing cuproptosis-like bacterial death against MRSA infections

The development of highly effective and low-toxicity antimicrobials is one of the important challenges for combat methicillin-resistant Staphylococcus aureus (MRSA) infections. Recent studies have found that cuproptosis-like bacterial death is an effective strategy to overcome drug resistance. Here, we designed and synthesized a valence in situ conversion of polyvalent copper oxide nanozymes (L-CuxO NPs), which modified L-arginine (L-Arg) to enhance antibacterial ability. In vitro studies showed that L-CuxO NPs can catalyze hydrogen peroxide (H2O2) to produce hydroxyl radical (•OH), mediate NO release, and continuously consume glutathione (GSH) through change of valence state of Cu2+ and Cu+. Importantly, copper overload in bacteria restricts the tricarboxylic acid (TCA) cycle, promoting bacterial cuproptosis-like death. In vivo results confirmed that L-CuxO NPs is not only effective in alleviating acute MRSA lung infection, but also accelerates the healing of MRSA infected wounds. Accordingly, this study provides a new idea for overcoming bacterial resistance and structural innovation of novel nanozymes.

The evolving role of mast cells in wound healing: insights from recent research and diverse models.

Chronic wounds significantly burden health care systems worldwide, requiring novel strategies to ease their impact. Many physiological processes underlying wound healing are well studied but the role of mast cells remains controversial. Mast cells are innate immune cells and play an essential role in barrier function by inducing inflammation to defend the host against chemical irritants and infections, among others. Many mast cell-derived mediators have proposed roles in wound healing; however, in vivo evidence using mouse models has produced conflicting results. Recently, studies involving more complex wound models such as infected wounds, diabetic wounds and wounds healing under psychological stress suggest that mast cells play critical roles in these processes. This review briefly summarizes the existing literature regarding mast cells in normal wounds and the potential reasons for the contradictory results. Focus will be placed on examining more recent work emerging in the last 5 years that explores mast cells in more complex systems of wound healing, including infection, psychological stress and diabetes, with a discussion of how these discoveries may inspire future work in the field.

Glucose-fueled cationic nanomotors for promoting the healing of infected diabetic wounds

Hyperglycemia-promoted bacterial infection will seriously exacerbate diabetic wounds, and its current clinical treatments are suffering from the adverse effects associated with off-target, bacterial resistance, and glycemic fluctuation. Herein, we present a kind of glucose-fueled cationic nanomotors capable of remarkably enhancing antibacterial efficacy, and thus expediting diabetic wound healing. The nanomotors have positively charged surfaces, and consist of mesoporous bowl-shaped polydopamine nanoparticles grafted with quaternized polymer brushes and coupled with glucose oxidase (GOx) and catalase (CAT). Stemming from the GOx-CAT cascade reaction in diabetic wound microenvironment, they can perform robust chemotactic motion towards both high glucose regions, where bacteria proliferation predominantly occurs, and elevated H2O2 levels, which bacterial metabolism produced. This enables the nanomotors to facilitate targeted migration towards bacteria-rich regions and simultaneous downregulation of glycemic levels, as well as to significantly enhance the electrostatic interaction between antibacterial components and bacteria. Consequently, the nanomotors exhibit amplified contact-killing effects of their attached cationic molecules, leading to an almost 10-fold enhancement in antibacterial efficacy compared to previous counterparts. The in vivo experiments approved that the nanomotors demonstrated the accelerated healing of infected diabetic wounds by S. aureus and biosafety. The results herein provide an insight into the clinical treatment of infected diabetic wounds.

Antibacterial and antioxidant supramolecular nanocomposite hydrogel dressings with angiogenesis and photo-thermal effect for multidrug‐resistant bacterial infected wound healing

Antioxidant, antibacterial and removable supramolecular nanocomposite hydrogel dressings with self-healing and tissue adhesiveness is highly anticipated for bacterial infected skin wound healing. A series of supramolecular nanocomposite hydrogels via quadruple hydrogen bond and coordination bonds (catechol-Cu) was developed based on ureidopyrimidinone-modified poly(glycerol sebacate)–co-poly(ethylene glycol)-g-catechol (PEGSDU) and mesoporous copper sulfide nanoparticle (CuS NPs) as antioxidative, antibacterial, removable and tissue-adhesive dressing to treat the MRSA-infected wound. The hydrogels exhibited good self-healing, free radical scavenging, photothermal sterilization performance, adhesiveness, and temperature-sensitive removability. In addition, the Cu2+ was able to be slowly released from the hydrogel, which can promote the proliferation and migration of fibroblasts. Hydrogels containing CuS NPs showed better wound repair efficiency than commercial TegadermTM films and hydrogels without CuS NPs in MRSA-infected skin wound, which was attributed to the good antibacterial property, improved collagen deposition, reduced inflammatory response, and improved new vessels formation, thereby significantly promoting the repair of wounds. In summary, this antioxidative and antibacterial supramolecular nanocomposite hydrogel dressing provide a new type of multi-functional dressing for the treatment of infected skin wounds.

NIR stimulated epigallocatechin gallate loaded polydopamine with enhanced antibacterial and ROS scavenging abilities for improved infectious wound healing

Infectious wound healing is complicated with and limited by infection and oxidative stress at the wound site. In recent years, various evidences suggest that nanozymes with multiple enzymatic activities have enabled the development of novel strategies for infectious wound healing. In this study, epigallocatechin gallate loaded polydopamine (P@E) was developed to act as a potent reactive oxygen species (ROS) scavenger for scavenging ROS, alleviating inflammatory responses, and promoting infectious wound healing. Combining with near infrared (NIR) irradiation, P@E presented excellent antibacterial ability of Escherichia coli (E. coli, 93.6%) and methicillin-resistant Staphylococcus aureus (MRSA, 87.6%). Specifically, P@E+NIR exhibited the most potent antioxidant, anti-inflammatory and cell proliferation behaviors through down-regulating intracellular ROS levels (81.9% and 94.3% for NIH3T3 and RAW264.7 respectively) and inducible nitric oxide synthase (iNOS) expression level (55.7%), and up-regulating the expression levels of arginase-1 (Arg-1, 71.4%), heat shock protein 70 (HSP70, 48.6%) and platelet endothelial cell adhesion molecule (CD31, 35.3%) compared to control group. Meanwhile, it also efficiently induced M2 directional polarization of lipopolysaccharide induced murine macrophages to achieve anti-inflammation, indicated by the down-regulation of CD86 (86.2%), and up-regulation of CD206 (85.6%). Significantly, it was also observed that P@E+NIR presented the excellent behaviors of inhibiting wound infection, alleviating wound inflammation, as well as promoting skin tissue repairing. Altogether, it has developed the strategy of using P@E combining with NIR irradiation for the synergistic enhanced healing of infectious skin wound, which can serve as a promising therapeutic strategy for its clinical treatment.

Engineering Injectable Coassembled Hydrogel by Photothermal Driven Chitosan-Stabilized MoS2 Nanosheets for Infected Wound Healing.

The application of enzyme-like molybdenum disulfide (MoS2) in tissue repair was confronted with stable dispersion, solubilization, and biotoxicity. Here, the injectable self-healing hydrogel was successfully designed using a step-by-step coassembly of chitosan and MoS2. Polyphenolic chitosan as a "structural stabilizer" of MoS2 nanosheets reconstructed well-dispersed MoS2@CSH nanosheets, which improved the biocompatibility of traditional MoS2, and strengthened its photothermal conversion and enzyme-like activities, guaranteeing highly efficient radical scavenging and antimicrobial properties. Furthermore, the polyphenol chitosan was employed again as a "molecular cross-linking agent" to form the injectable NIR-responsive MoS2@CSH hydrogel by accelerating hydrogen-bond interaction among chitosan and the multicross-linking reaction among polyphenols. The rapid self-healing ability was conducive to wound closure and dynamic adaptability. An experimental study on infected wound healing demonstrated that MoS2@CSH hydrogel could substantially eradicate bacteria and accelerate the angiogenesis of infected wounds. The photothermal-driven coassembly of MoS2 and polycation provided an alternative strategy for infected wound healing.

Multifunctional hydrogel based on polyvinyl alcohol/chitosan/metal polyphenols for facilitating acute and infected wound healing

Multifunctional hydrogel based on polyvinyl alcohol/chitosan/metal polyphenols for facilitating acute and infected wound healing

Recombinant Keratin-Chitosan Cryogel Decorated with Gallic Acid-Reduced Silver Nanoparticles for Wound Healing.

Wound healing is a complex physiological process that can be roughly divided into four stages: hemostasis, inflammation, proliferation, and remodeling. Conventional wound dressings often fail to meet the diverse needs of these healing stages due to their limited functionality. Cryogels, however, possess several attractive properties, such as large, interconnected pores, good mechanical strength, and ease of modification, making them suitable for developing advanced dressings with multiple functions. In this study, we developed a multifunctional cryogel dressing, with biocompatible polysaccharides as the main component, designed to provide a breathable, moist, and antibacterial microenvironment for chronic infected wounds, thereby promoting wound healing. Recombinant keratin 31 (RK31) was combined with chitosan (CS) to produce a CS/RK31 cryogel, referred to as CK. Gallic acid-reduced silver nanoparticles (GA/Ag NPs) were incorporated as the active antibacterial component to create the CS/K31@GA/Ag cryogel, known as CKGA. The cryogel was characterized using scanning electron microscopy (SEM) and a universal testing machine, and its biocompatibility was assessed in vitro. The dynamic hemostatic performance of the cryogel was evaluated with a rat tail amputation bleeding model. Additionally, the antibacterial effects of the cryogel against Staphylococcus aureus and Escherichia coli were tested using agar diffusion assays and turbidimetry. The antioxidant capacity of the CKGA cryogel was also measured in vitro. Finally, the cryogel's ability to promote wound healing was tested in an SD rat model of infected wounds. Characterization results showed that the CKGA cryogel features an interpenetrating porous network structure and exhibits excellent mechanical properties, with a swelling rate of up to 1800%. Both in vitro and in vivo experiments confirmed that the cryogel has good biocompatibility, effectively absorbs exudates, and rapidly stops bleeding. The addition of GA/Ag NPs provided significant antibacterial effects, achieving an inhibition rate of over 99.9% against both S. aureus and E. coli. Furthermore, CKGA cryogels demonstrated a strong scavenging capacity for ROS in a dose-dependent manner. Studies using the SD rat infected wound model showed that the cryogel effectively inhibited bacterial proliferation on wound surfaces, reduced local tissue inflammation, and promoted the healing of infected wounds. The multifunctional cryogel, with its rapid hemostatic, antibacterial, and antioxidant properties, as well as its ability to promote cell proliferation, could be widely used as a wound dressing for the healing of bacterial infections.

Injectable Oxygen-Carrying Microsphere Hydrogel for Dynamic Regulation of Redox Microenvironment of Wounds.

The delayed healing of infected wounds can be attributed to the increased production of reactive oxygen species (ROS) and consequent damages to vascellum and tissue, resulting in a hypoxic wound environment that further exacerbates inflammation. Current clinical treatments including hyperbaric oxygen therapy and antibiotic treatment fail to provide sustained oxygenation and drug-free resistance to infection. To propose a dynamic oxygen regulation strategy, this study develops a composite hydrogel with ROS-scavenging system and oxygen-releasing microspheres in the wound dressing. The hydrogel itself reduces cellular damage by removing ROS derived from immune cells. Simultaneously, the sustained release of oxygen from microspheres improves cell survival and migration in hypoxic environments, promoting angiogenesis and collagen regeneration. The combination of ROS scavenging and oxygenation enables the wound dressing to achieve drug-free anti-infection through activating immune modulation, inhibiting the secretion of pro-inflammatory cytokines interleukin-6, and promoting tissue regeneration in both acute and infected wounds of rat skins. Thus, the composite hydrogel dressing proposed in this work shows great potential for dynamic redox regulation of infected wounds and accelerates wound healing without drugs.

Dynamic polysaccharide/platelet-rich plasma hydrogels with synergistic antibacterial activities for accelerating infected wound healing

Platelet-rich plasma (PRP) has been recognized as an effective therapy in regenerative medicine and surgery, which can reduce the risk of antibiotic abuse and promote the healing of infected wounds. Recent advances in PRP-based treatments have focused on the controlled release of growth factors in PRP with biocompatible hydrogels and antimicrobial promotion by introducing hydrogel components or antibiotics, while the inherent antimicrobial activity of PRP is mostly neglected or sacrificed. Here, we demonstrate the combination of an antimicrobial polysaccharide, carboxymethyl chitosan, and PRP to construct an antimicrobial hydrogel via dynamic bonding with oxidized chondroitin sulfate. Significant inhibitory effects against Staphylococcus aureus and Escherichia coli (95 % of inhibition rate) are achieved through the synergistic contributions of the polysaccharide and PRP. Additionally, the resulting hydrogel promotes the migration of NIH-3T3 fibroblasts and collagen deposition by approximately 1.7 and 1.8 times, respectively, thereby accelerating the healing process of infected wounds. This work may bring new perspectives for potent applications of PRP-based hydrogel dressings for antibiotic-free management of infected wounds.

Caffeic acid-mediated photodynamic multifunctional hyaluronic acid-gallic acid hydrogels with instant and enduring bactericidal potency accelerate bacterial infected wound healing

The emergence of drug-resistant bacteria poses significant challenges in wound treatment. Antimicrobial photodynamic therapy has emerged as an effective approach to eliminating bacteria by inducing oxidative stress without causing drug resistance. Here, we developed a natural hyaluronic acid (HA)-gallic acid (GA) conjugation-based hydrogel combined with herbal photosensitizer-caffeic acid (CA), which exhibits self-healing ability, shape adaptability, biodegradability, and robust tissue adhesion. Under exposure to 400 nm light, caffeic acid acts as a photosensitizer, generating reactive oxygen species and oxidative damage to bacterial cell membranes. Furthermore, the presence of GA and CA displayed a continuous inhibitory effect on bacterial growth, along with antioxidant properties that promote wound healing even after the cessation of light exposure. The antibacterial mechanism of the HA-GA/CA hydrogel against MRSA, S. aureus, and E. coli was investigated through various assays measuring ATP levels, Zeta potential, hydroxyl radicals (·OH) generated by light irradiation, and biofilm clearance rate. Additionally, hydrogel's application in treating MRSA-infected wounds in mice under light irradiation demonstrated rapid wound-healing effects and biocompatibility. Overall, HA-GA/CA hydrogel provides a sustainable, antibiotic-free alternative for treating MRSA-infected wounds.

Curcumin-Loaded Nanocomposite Hydrogel Dressings for Promoting Infected Wound Healing and Tissue Regeneration

Curcumin-Loaded Nanocomposite Hydrogel Dressings for Promoting Infected Wound Healing and Tissue Regeneration

Gellan gum-based multifunctional hydrogel with enduring sterilization and ROS scavenging for infected wound healing

The progression of severe skin injury healing can be easily impeded by bacterial infections and the resultant overproduction of reactive oxygen species (ROS) within the wound microenvironment. In this study, we developed a multifunctional antibacterial hydrogel by integrating gallium ion-tannic acid and polydopamine particles into gellan gum via a facile heat-cooling process. By harnessing the synergistic effects of polydopamine for short-term photothermal therapy and gallium ion for long-term chemotherapy, the hydrogel obtained shows outstanding antibacterial activities. Sustained release of gallium ion and tannic acid ensures a prolonged sterilization along with ROS-scavenging benefits. Moreover, this hydrogel demonstrates superior cytocompatibility, hemostatic properties, as well as capabilities including promoting cell migration, and adsorption to bacterial cells and toxin. The therapeutic efficacy of the hydrogel was validated using a mouse model of MRSA-induced cutaneous infections. Overall, this work introduces a straightforward yet highly efficient multifunctional hydrogel platform that combines synergetic antibacterial actions, ROS scavenging, and hemostasis to enhance the healing of bacteria-associated wounds.

WITHDRAWN: Natural outer membrane vesicles from Serratia marcescens for inherent and biocompatible anti-microbial photodynamic therapy

It is a common pursuit to create an efficient photodynamic therapy (PDT), as a potential alternative to antibiotics. In this research, we first discovered and acquired the natural outer membrane vesicles (OMVs) from Serratia marcescens for effective photodynamic inactivation of both bacteria and fungi. The native OMVs was prepared from the culture media of S. marcescens, which inherently contained a photosensitizer named prodigiosin (PG). The negatively-charged, green-emitting OMVs exhibited improved water-solubility and increased singlet oxygen generation, as compared to free PG, resulting in notably enhanced antibacterial PDT effect towards microbes with appealing biosafety in vitro. Upon binding to the microbial cells through hydrophobic interaction, OMVs enriched PG in the microbic cells, destroying cell membrane structure and DNA and causing protein leakage under light irradiation, leading to microbial cell death. More importantly, OMVs could notably enhance infected wound healing and antibacterial effectiveness with satisfactory biocompatibility in mice. Collectively, the naturally-occurring PG-containing OMVs offers a novel modality for effective antimicrobial PDT, benefiting the treatment of microbial infection in clinic.

Self-assembled near-infrared-photothermal antibacterial Hericium erinaceus β-glucan/tannic acid/Fe (III) hydrogel for accelerating infected wound healing

Bacterial infection severely hinders skin wound healing, highlighting the critical application value of developing antibacterial and anti-inflammatory hydrogel dressings. In this work, we focused on β-glucan from Hericium erinaceus as the research object, and proposed a solvent-induced combined temperature manipulation technique to trigger multilevel self-assembly of β-glucan. Furthermore, we incorporated green synthesized near-infrared photosensitizer tannic acid/iron (III) complex into the system. A hydrogel with exceptional antibacterial properties, capable of responding to near-infrared photothermal stimuli while exhibiting remarkable stiffness and structural consistency, was successfully synthesized. Under near-infrared radiation, HEBG/TA/Fe hydrogels produced local hyperthermia and exhibited excellent antibacterial activity against bacteria-infected wounds. Moreover, the HEBG/TA/Fe hydrogel demonstrates its ability to regulate cytokines by effectively inhibiting the production of inflammatory mediators TNF-α and IL-6, while simultaneously enhancing the expression of cell proliferation factor KI-67 and markers associated with angiogenesis such as CD31 and α-SMA. Notably, the results of tissue staining revealed that NIR + HEBG/TA/Fe5 hydrogel could effectively promoting granulation and vascularization, improving collagen deposition in infected wounds thereby accelerating the healing process. These findings indicate that mixed hydrogels exhibit potential as viable options for the treatment of bacterial infections.

Lactobacillus rhamnosus GG and Bifidobacterium animalis subsp. lactis BB-12 promote infected wound healing via regulation of the wound microenvironment.

Infected wounds can result in complex clinical complications and delayed healing, presenting a significant global public health challenge. This study explored the effects of topical application of two probiotics, Lactobacillus rhamnosus GG (LGG) and Bifidobacterium animalis subsp. lactis BB-12, on the microenvironment of infected wounds and their impact on wound healing. LGG and BB-12 were applied separately and topically on the Staphylococcus aureus (S. aureus)-infected skin wounds of the rat model on a daily basis. Both probiotics significantly accelerated wound healing, demonstrated by enhanced granulation tissue formation and increased collagen deposition, with BB-12 showing superior efficacy. LGG and BB-12 both effectively inhibited neutrophil infiltration and decreased the expression of pro-inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Notably, BB-12 markedly reduced IL-6 levels, while LGG significantly lowered TNF-α, transforming growth factor-β (TGF-β) and vascular endothelial growth factor (VEGF). Additionally, both probiotics promoted macrophage polarization towards the anti-inflammatory M2 phenotype. Microbiota analysis revealed that LGG and BB-12 significantly decreased the abundance of pathogenic bacteria (e.g. Staphylococcus and Proteus) and increased the proportion of beneficial bacteria (e.g. Corynebacterium). Particularly, BB-12 was more effective in reducing Staphylococcus abundance, whereas LGG excelled in promoting Corynebacterium growth. These findings suggest the ability of LGG and BB-12 to modulate the wound microenvironment, enhance wound healing and provide valuable insights for the management of infected wounds.

Multifunctional NIR-II nanoplatform for disrupting biofilm and promoting infected wound healing

Healing wounds presents a significant challenge due to bacterial biofilm infections and the inherent drug resistance of these biofilms. This report introduces a multifunctional nanoplatform (NPs) designed to combat wound biofilm infections using NIR-II photothermal therapy. The NPs are self-assembled from amphiphilic polymers (AP) to encapsulate photothermal polymers (PT) through classic electrostatic interactions. Importantly, these NPs are electrically neutral, which enhances their ability to penetrate biofilms effectively. Once inside the biofilm, the NPs achieve complete thermal ablation of the biofilm under NIR-II laser irradiation. Additionally, when exposed to laser and the GSH microenvironment, the NPs exhibit strong photothermal effects and self-degradation capabilities. In vitro tests confirm that the NPs have excellent antibacterial and anti-biofilm properties against methicillin-resistant Staphylococcus aureus (MRSA). In vivo studies demonstrate that the NPs can efficiently clear wound biofilm infections and promote wound healing. Notably, the NPs show superior photothermal effects under NIR-II laser irradiation compared to NIR-I lasers. In summary, the developed NPs serve as an integrated diagnostic and therapeutic nano-antimicrobial agent, offering promising applications for biofilm wound infections and wound healing.

A multifunctional hydrogel dressing loaded with antibiotics for healing of infected wound

Wound bacterial infections can significantly delay the healing process and even lead to fetal sepsis. There is a need for multifunctional dressings that possess antibacterial property, tissue adhesive property, self-healing capability, and biocompatibility to effectively treat bacteria-infected wound. In this study, we report a dual dynamically crosslinked hydrogel, OHA-PBA/PVA/Gen, which incorporates the antibiotic gentamicin (Gen) as a dynamic crosslinker. The hydrogel is formed through the formation of Schiff base bonds between phenylboronic acid-grafted oxidized hyaluronic acid (OHA-PBA) and Gen, as well as boronic acid ester bonds between OHA-PBA and polyvinyl alcohol (PVA). This unique composition imparts tissue adhesiveness, injectability and self-healing property to the hydrogel. The hydrogel also exhibits pH-responsive antibiotic release behavior due to the acid-responsive dissociation of Schiff base bonds. As a result, it demonstrates strong antibacterial activity against both Gram-positive bacteria S. aureus and Gram-negative bacteria E. coli through contact killing and diffusion killing mechanisms. Importantly, the OHA-PBA/PVA/Gen hydrogel avoids incorporation of toxic small molecular crosslinking agents, and all the components of the hydrogel are biocompatible, ensuring its biosafety. In a S. aureus-infected wound mouse model, this hydrogel effectively eradicated bacteria and promoted angiogenesis, leading to significantly accelerated wound healing. These results highlight the potential of the dual dynamically crosslinking hydrogel OHA-PBA/PVA/Gen as a multifunctional wound dressing for the treatment of bacteria-infected wound.

Optimization In-vitro Evaluation and Scratch Wound Healing Assay of Hexacosane Topical Gel for Wound Healing.

The compounds hexocosane, a sterol hydrocarbon identified in the dichloromethane extract and isolated from marine sponges of the genus Agelas. Hexocosane possesses anti-inflammatory and antioxidant activities that reduce inflammation and oxidative stress, they promote cell proliferation and angiogenesis, facilitating tissue regeneration and repair. This multifaceted approach makes this secondary metabolite a promising candidate for developing advanced wound care therapies that effectively address both infection control and wound healing. On the basis of viscosity, in-vitro release, and skin retention, optimal gel formulations were developed with hexocosane (1%) to achieve the best results. A response surface methodology Box-Behnken design was applied on three factors and three levels [carbopol 940 (1, 1.25 and 1.5%), hydroxypropyl methylcellulose (HPMC) (1,1.5 and 1.2%), and propylene glycol (0–1–1.5% and 1–2%, respectively] using three factors and three levels. In order to assess the spreadability and viscosity, a glass slide and a Brookfield viscometer were used. An assay was conducted to determine how topical gel affects wound healing. The optimization study of the gel formulation, involving variations in adhesive polymer (Carbopol 940), release retarding polymer (HPMC K4M), and penetration enhancer (Surfactant), has identified three noteworthy formulations (F4, F8 & F14). Each formulation is characterized by specific attributes such as viscosity, in-vitro drug release, and skin retention. All samples were found to elicit significant wound healing efficacy as evidenced from the representative photomicrographs. The maximum efficacy was elicited by formulation (F4) with 100 mg drug at the time duration of 36 hours.