Wound Microenvironment Research Articles

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.

Novel thermal-regulation and temperature-responsive controlled-release fibermats with porous sheath-core structures

Active regulation of the wound microenvironment as well as on-demand drug delivery have proven to be effective ways to accelerate wound healing, and the development of advanced textiles offers feasibility. A novel fibermat with thermal-regulation and temperature-responsive controlled-release properties was fabricated by using coaxial electrospinning, consisting of porous sheath-core polylactic acid (PLA)/ polyethylene glycol (PEG) fibers. The pore generation in the PLA shell of the PLA/PEG fiber was achieved through a phase separation method. The fiber diameter and pore size of the obtained fiber could be controlled by adjusting the flow rate of shell solution during electrospinning. Increasing the flow rate of shell solution form 0.8 mm·min-1 to 1.4 mm·min-1 resulted in an increase in fiber diameter from 4.8 μm to 15.3 μm, and an increase in pore size from 59.4 nm to 305.6 nm. Due to the high enthalpy and coverage of the PEG core, the PLA/PEG fibermats exhibited excellent thermal-regulation performance, with a surface temperature that was 2.5 °C- 5.6 °C lower than that of pure PLA fibermat during heating process. On-demand release was achieved by directly controlling the core layer of the coaxial PLA/PEG fiber through temperature. The hydrophobicity of the PLA shell initially reduced burst release, followed by increased release as the temperature reached the phase transition temperature of PEG. Furthermore, this advanced fibermat demonstrated desirable moisture permeability, appropriate mechanical properties and good thermal stability, indicating broad potential applications in medical and health fields.

Immunomodulation of Macrophages in Diabetic Wound Individuals by Structurally Diverse Bioactive Phytochemicals

Diabetes-related ulcers and slow-healing wounds pose a significant health risk to individuals due to their uncertain causes. Mortality rates for diabetes foot ulcers (DFUs) range from 10% after 16 months to 24% after five years. The use of bioactive phytochemicals can play a key role in healing wounds in a predictable time. Recent literature has demonstrated that various natural substances, including flavonoids, saponins, phenolic compounds, and polysaccharides, play key roles at different stages of the wound-healing process through diverse mechanisms. These studies have categorized the compounds according to their characteristics, bioactivities, and modes of action. In this study, we evaluated the role of natural compounds derived from plant sources that have been shown to play a crucial role in immunomodulation. Macrophages are closely involved in immunomodulation within the wound microenvironment and are key players in efferocytosis, inflammation resolution, and tissue regeneration, all of which contribute to successful wound healing. Phytochemicals and their derivatives have shown capabilities in immune regulation, including macrophage migration, nitric oxide synthase inhibition, lymphocyte and T-cell stimulation, cytokine activation, natural killer cell enhancement, and the regulation of NF-κβ, TNF-α, and apoptosis. In this review, we have studied the role of phytochemicals in immunomodulation for the resolution of diabetic wound inflammation.

CD9 promotes TβR2–TβR1 association driving the transition of human dermal fibroblasts to myofibroblast under hypoxia

BackgroundDuring wound healing, fibroblast to myofibroblast transition is required for wound contraction and remodeling. While hypoxia is an important biophysical factor in wound microenvironment, the exact regulatory mechanism underlying hypoxia and fibroblast-to-myofibroblast transition remains unclear. We previously found that tetraspanin CD9 plays an important role in oxygen sensing and wound healing. Herein, we investigated the effects of physiological hypoxia on fibroblast-to-myofibroblast transition and the biological function and mechanism of CD9 in it.MethodsHuman skin fibroblasts (HSF) and mouse dermis wounds model were established under physiological hypoxia (2% O2). The cell viability and contractility of HSF under hypoxia were evaluated by CCK8 and collagen gel retraction, respectively. The expression and distribution of fibroblast-to-myofibroblast transition markers and CD9 in HSF were detected by Western blotting and immunofluorescence. CD9 slicing and overexpressing HSFs were constructed to determine the role of CD9 by small interfering RNA and recombinant adenovirus vector. The association of TβR2 and TβR1 was measured by immunoprecipitation to explore the regulatory mechanism. Additionally, further validation was conducted on mouse dermis wounds model through histological analysis.ResultsEnhanced fibroblast-to-myofibroblast transition and upregulated CD9 expression was observed under hypoxia in vitro and in vivo. Besides, reversal of fibroblast-to-myofibroblast transition under hypoxia was observed when silencing CD9, suggesting that CD9 played a key role in this hypoxia-induced transition. Moreover, hypoxia increased fibroblast-to-myofibroblast transition by activating TGF-β1/Smad2/3 signaling, especially increased interaction of TβR2 and TβR1. Ultimately, CD9 was determined to directly affect TβR1–TβR2 association in hypoxic fibroblast.ConclusionCollectively, these findings suggest that CD9 promotes TβR2–TβR1 association, thus driving the transition of human dermal fibroblasts to myofibroblast under hypoxia.

Mouse Wound Models and Preparation of Single-Cell Suspensions.

The management of acute full-thickness skin injuries presents a considerable challenge in clinical practice. The complexity of wound healing, involving diverse cell populations and the intricate wound microenvironment, complicates the assessment of treatment effects and their interactions using traditional experimental approaches. Single-cell transcriptome sequencing technology has revolutionized the understanding of cellular functions and mechanisms during wound healing. However, the variability in methodologies for creating mouse wound models introduces confounding factors that can impact experimental results. Furthermore, the process of dissociating mouse skin tissues presents significant technical hurdles, highlighting the critical need for high-quality single-cell suspensions for accurate transcriptome sequencing. Therefore, this study aims to optimize the construction of mouse wound models to accurately assess changes in wound closure while eliminating variables that could influence the subsequent sequencing result. Additionally, the preparation of single-cell suspensions was performed using both enzyme preparation protocols and test kit protocols to optimize cost and effectiveness.

Assessment and Monitoring of the Wound Micro-Environment in Chronic Wounds Using Standardized Wound Swabbing for Individualized Diagnostics and Targeted Interventions.

Background/Objectives: Patient-specific diagnostic and therapeutic approaches are important in the care of people with chronic wounds. The heterogeneity of underlying disease profiles and the diversity of the wound micro-environment make generalized approaches difficult. While high-throughput molecular diagnostic methods are increasingly widespread and available, the analysis of objective biomolecular disease patterns has not found its way into everyday wound management. The aim of this study is to evaluate the use of wound swab samples for the analysis of biomarkers and disease patterns in people with chronic wounds. Methods: A sample cohort from the multicenter "Wound-BIOME" project was analyzed. The project aims to comprehensively investigate the local micro-environment of chronic wounds of various entities, healing tendencies and regeneration stages at the biomolecular level. A sample collection and handling protocol suitable for everyday use was tested and evaluated regarding feasibility for multiplex immunoassay, proteomics, small RNA sequencing (miRNA) and metagenome analyses (microbiomics). Results: It could be shown that standard wound swabs are well-suited for the analysis of the complex wound micro-environment using various high-throughput methods. Despite the sample heterogeneity, the quality was adequate to analyze biomolecular patterns. Conclusions: Initial analyses of protein signatures, microbial wound communities and miRNA patterns show promising results for future individualized diagnostics and targeted interventions.

Multi-drug loaded electrospinning nanofibers promote healing with less scar formation by remodeling wound microenvironment via modulating TGF-β1/Smads/EN1 signal

Excessive scar formation following severe burn injury, deep trauma or surgical incision is a critical complication and still is a thorny clinical troublesome, which are mainly characterized by the disordered microenvironment during wound healing. A specialty wound dressing remodeling wound microenvironment is urgently desired. Herein, we fabricate a multi-drug loaded nanofiber delivery system for promoting healing and mitigating scar formation by modulating TGF-β1/Smads/EN1 signal, which encapsulates free rhubarb anthraquinones (RAs) and honey in polyvinylpyrrolidone core layer, and borneol in cellulose acetate shell layer by coaxial electrospinning. The nanofibers exhibited a linear morphology with smooth surface, nanometer in diameter, high porosity, ideal water vapor transmission rate and wettability, a biphasic release profile of RAs with the enhanced dissolution under the synergism of honey and borneol. Furthermore, this nanofiber wound dressing possesses the desired biocompatibility and pro-migration effects on HSF cells and HUVEC, long-term effects against Methicillin-resistant Staphylococcus aureus (MRSA) in vitro, remarkably, promoted healing of the MRSA-infected postsurgical wound and severe burn injury characterized by enhanced closure rates and more hair follicles, and mitigated scar formation characterized by less dermal thickness and more organized collagen arrangement by decreasing the expressions of TGF-β1, Smads2/3 and EN1, and increasing Smad7 expression in wound tissues. Collectively, the findings indicate that the multi-drug loaded nanofiber wound dressing can serve as a promising candidate for the application of MRSA-infected wound and severe burn injury healing.

A ROS‐Responsive Lipid Nanoparticles Release Multifunctional Hydrogel Based on Microenvironment Regulation Promotes Infected Diabetic Wound Healing

AbstractThe continuous imbalance of the diabetic wound microenvironment is an important cause of chronic nonhealing, which manifests as a vicious cycle between excessive accumulation of reactive oxygen species (ROS) and abnormal healing. Regulating the microenvironment by suppressing wound inflammation, oxidative stress, and bacterial infection is a key challenge in treating diabetic wounds. In this study, ROS‐responsive hydrogels are developed composed of silk fibroin methacrylated (SFMA), modified collagen type III (rCol3MA), and lipid nanoparticles (LNPs). The newly designed hydrogel system demonstrated stable physicochemical properties and excellent biocompatibility. Moreover, the release of antimicrobial peptide (AMP) and puerarin (PUE) demonstrated remarkable efficacy in eradicating bacteria, regulating inflammatory responses, and modulating vascular functions. This multifunctional hydrogel is a simple and efficient approach for the treatment of chronic diabetic infected wounds and holds tremendous potential for future clinical applications.

Cu2O-SnO2-PDA heterozygous nanozyme doped hydrogel mediated conglutinant microenvironment regulation for wound healing therapy

Bacterial infection significantly hinders the wound healing process. Overuse of antibiotics has led to the rise of drug resistance in bacteria, making the development of smart medical dressings that promote wound healing without antibiotics, a critical need. In this study, Cu₂O-SnO₂-PDA (PCS) nanoenzymes with Fenton-like activity and high photothermal conversion efficiency were developed. These nanoenzymes were then incorporated into a hydrogel through cross-linking of acrylamide (AM) and N-[Tris-(hydroxymethyl)methyl] acrylamide (THMA), forming a tough, highly-adhesive, and self-healing composite hydrogel (AT/PCS) with antimicrobial properties. The AT/PCS hydrogel exhibits excellent mechanical strength and adhesion, facilitating increased oxygen levels and strong adherence to the wound site. Moreover, it effectively regulates the wound microenvironment by combining synergistic chemodynamic therapy (CDT) and photothermal therapy (PTT) for antibacterial treatment. The AT/PCS hydrogel enhances collagen deposition and expedites wound healing in a rat model, largely due to its potent antibacterial properties.

Radial Egg White Hydrogel Releasing Extracellular Vesicles for Cell Fate Guidance and Accelerated Diabetic Skin Regeneration.

Topology and bioactive molecules are crucial for stimulating cellular and tissue functions. To regulate the chronic wound microenvironment, mono-assembly technology is employed to fabricate a radial egg white hydrogel loaded with lyophilized adipose tissue-extracellular vesicles (radial EWH@L-EVs). The radial architecture not only significantly modified the gene expression of functional cells, but also achieved directional and controlled release kinetics of L-EVs. Through the synergy of topographical and inherent bioactive cues, radial EWH@L-EVs effectively reduced intracellular oxidative stressand promoted the polarization of macrophages toward an anti-inflammatory phenotype during the inflammatory phase. Afterward, radial EWH@L-EVs facilitated the centripetal migration and proliferation of fibroblasts and endothelial cells as the wound transitioned to the proliferative phase. During the latter remodeling phase, radial EWH@L-EVs accelerated the regeneration of granulation tissue, angiogenesis, and collagen deposition, thereby promoting the reorganization chronic wound. Compared with the gold standard collagen scaffold, radial EWH@L-EVs actively accommodated the microenvironment via various functions throughout all stages of diabetic wound healing. This can be attributed to the orientation of topological structures and bioactive molecules, which should be considered of utmost importance in tissue engineering.

All-natural hydrogel composed of carboxymethyl chitosan and oxidized dextran for promoting wound healing by immune-microenvironment regulation

Chronic wound treatment has always been a major clinical challenge owing to complex and dynamic wound microenvironment. The design and development of effective management with antimicrobial activity, ROS-scavenging and regulation immune response is vital for tissue repair. Herein, we developed puerarin (PUE) loaded a double network hydrogel consisting of methacrylated carboxymethyl chitosan and oxidized dextran with Schiff base and photo-crosslinking reaction. The composite hydrogel presented fast self-healing and outstanding compressive performance to bear deformation. The hydrogel exhibits a cumulative drug release pattern with biphasic release, which offers great potential for accelerating tissue healing at all stages of wounding. In addition, the hydrogel has good biocompatibility, antimicrobial ability and tube-forming ability in vitro. The full-thickness skin wound model of Staphylococcus aureus infection showed that the hydrogel dressing can accelerate tissue repair. This study demonstrated that the design of combing natural biomacromolecules with traditional Chinese medicine could be severed as a promising candidate biomaterial for skin tissue recovery.

Non-releasing poly (ionic liquid) based hydrogel accelerates diabetic wound healing

Persistent bacterial colonization, abnormal inflammatory responses, and impaired angiogenesis pose significant challenges to effective wound repair, particularly in diabetic wounds. Employing exogenous bioactive substances in wound dressings is a recognized approach to dynamically respond to the wound microenvironment and accelerate the repair process. However, this strategy can lead to the development of drug resistance and induce further tissue damage. To address these challenges, we are synthesizing a novel hydrogel for diabetic wound treatment using functional poly (ionic liquid) and modified dextran. The hydrogel is characterized by its excellent tissue adhesion, exceptional self-healing capacity, and substantial compressive deformation. It exhibits broad antibacterial activity, reduces the expression of pro-inflammatory cytokines and enhances the healing in diabetic wounds. Its efficacy is superior to that of the positive control group. This innovative non-releasing hydrogel presents as a promising alternative to conventional antibiotics, offering significant potential for the treatment and healing of diabetic chronic wounds.

Intelligent Carbon Dots with Switchable Photo-Activated Oxidase-Mimicking Activity and pH Responsive Antioxidant Activity Adaptive to the Wound Microenvironment for Selective Antibacterial Therapy.

Intelligent antibacterial agent with controllable activities adaptive to the wound microenvironment is appealing to reduce drug resistance and enhance antibacterial efficiency. In this study, celery is chosen as the carbon source to construct celery-based carbon dots (CECDs) with double activities, i.e., reactive oxygen species (ROS)-production and ROS-clearance activities. The ROS-production capability of CECDs is dependent on the oxidase (OXD)-mimicking activity, which is only photo-activated and thus artificially controlled by light to avoid the production of excess ROS. Meanwhile, the optimal OXD-mimicking activity occurrs at the pH of 5, close to microenvironmental pH at the bacterial infection site, which will enhance the antibacterial efficacy. On the other hand, CECDs exhibit the antioxidant activity at the neutral or weak alkaline pH, which will assist the healing of the wound. Thus, the conversion of ROS-production and ROS-clearance ability of CECDs can be dynamically and intelligently switched automatically with microenvironmental pH at different stages of treatment (from acid to neutral/weak basic). The proposed CECDs exert adorable selective antibacterial activity against Gram-positive bacteria and satisfactory therapeutic effect on bacteria infected mice. This study paves a new avenue to design the intelligent antibacterial nanoagent sensitive to the infected microenvironmental condition, reducing drug resistance and assisting precise medicine.

Efficacy and safety of autologous whole blood clot in diabetic foot ulcers: a randomised controlled trial.

Diabetic foot ulcers (DFUs) present a significant global health challenge, resulting in high morbidity and economic costs. Current available treatments often fail to achieve satisfactory healing rates, highlighting the need for novel therapies. This study evaluated the safety and efficacy of a novel autologous whole blood clot (AWBC)-a blood-based, biodegradable provisional matrix-in conjunction with standard of care (SoC) when compared to SoC alone in the treatment of hard-to-heal DFUs. A multicentre, prospective, blinded assessor, randomised controlled trial was conducted at 16 sites across the US, South Africa and Turkey. A cohort of patients with hard-to-heal DFUs was enrolled and randomised to either the AWBC group or the control group. The primary endpoint was complete wound closure at 12 weeks, while secondary endpoints included time to heal and percentage area reduction (PAR) at four and eight weeks. Data were analysed using both intention-to-treat (ITT) and per-protocol (PP) populations. The cohort included 119 patients. AWBC treatment resulted in a significantly higher healing rate compared to the control in both ITT (41% versus 15%, respectively; p=0.002) and PP populations (51% versus 18%, respectively; p=0.0075). AWBC treatment also resulted in a shorter mean time to heal and higher durability of wound closure. Safety analysis showed a similar incidence of adverse events (AEs) between groups, with no device-related AEs. The AWBC system, by modulating the wound microenvironment and providing a functional extracellular matrix, offered a promising new approach to treating hard-to-heal DFUs, demonstrating superior healing outcomes compared to SoC alone in this study.

Multifunctional hydrogel dressing promotes wound healing by reprogramming the infection-related wound microenvironment

Multifunctional hydrogel dressing promotes wound healing by reprogramming the infection-related wound microenvironment

In Situ Triggered Self-Contraction Bioactive Microgel Assembly Accelerates Diabetic Skin Wound Healing by Activating Mechanotransduction and Biochemical Pathway.

Chronic nonhealing skin wounds, characterized by reduced tissue contractility and inhibited wound cell survival under hyperglycemia and hypoxia, present a significant challenge in diabetic care. Here, an advanced self-contraction bioactive core-shell microgel assembly with robust tissue-adhesion (SMART-EXO) is introduced to expedite diabetic wound healing. The SMART-EXO dressing exhibits strong, reversible adhesion to damaged tissue due to abundant hydrogen and dynamic coordination bonds. Additionally, the core-shell microgel components and dynamic coordination bonds provide moderate rigidity, customizable self-contraction, and an interlinked porous architecture. The triggered in situ self-contraction of the SMART-EXO dressing enables active, tunable wound contraction, activating mechanotransduction in the skin and promoting the optimal fibroblast-to-myofibroblast conversion, collagen synthesis, and angiogenesis. Concurrently, the triggered contraction of SMART-EXO facilitates efficient loading and on-demand release of bioactive exosomes, contributing to re-epithelialization and wound microenvironment regulation in diabetic mice. RNA-seq results reveal the activation of critical signaling pathways associated with mechanosensing and exosome regulation, highlighting the combined biomechanical and biochemical mechanisms. These findings underscore SMART-EXO as a versatile, adaptable solution to the complex challenges of diabetic wound care.

Damage-induced basal epithelial cell migration modulates the spatial organization of redox signaling and sensory neuron regeneration.

Epithelial damage leads to early reactive oxygen species (ROS) signaling, which regulates sensory neuron regeneration and tissue repair. How the initial type of tissue injury influences early damage signaling and regenerative growth of sensory axons remains unclear. Previously we reported that thermal injury triggers distinct early tissue responses in larval zebrafish. Here, we found that thermal but not mechanical injury impairs sensory axon regeneration and function. Real-time imaging revealed an immediate tissue response to thermal injury characterized by the rapid Arp2/3-dependent migration of keratinocytes, which was associated with tissue scale ROS production and sustained sensory axon damage. Isotonic treatment was sufficient to limit keratinocyte movement, spatially restrict ROS production, and rescue sensory neuron function. These results suggest that early keratinocyte dynamics regulate the spatial and temporal pattern of long-term signaling in the wound microenvironment during tissue repair.

Temperature-responsive self-contraction nanofiber/hydrogel composite dressing facilitates the healing of diabetic-infected wounds

Bacterial infections and long-term inflammation cause serious secondary damage to chronic diabetic wounds and hinder the wound healing processes. Currently, multifunctional hydrogels have shown promising effects in chronic wound repair. However, traditional hydrogels only keep the wound moist and protect it from bacterial infection, and cannot provide mechanical force to contract the wound edges to achieve facilitated wound closure. Here, an asymmetric composite dressing was created by combining biaxially oriented nanofibers and hydrogel, inspired by the double-layer structure of the traditional Chinese medicinal plaster patch, for managing chronic wounds. Specifically, electrospun Poly-(lactic acid-co-trimethylene carbonate) (PLATMC) nanofibers and methacrylate gelatin (GelMa) hydrogel loaded with Epinecidin-1@chitosan (Epi-1@CS) nanoparticles are assembled as the temperature-responsive self-contracting nanofiber/hydrogel (TSNH) composite dressing. The substrate layer of PLATMC nanofibers combines topological morphology with material properties to drive wound closure through temperature-triggered contraction force. The functional layer of GelMa hydrogel is loaded with Epi-1@CS nanoparticles that combine satisfactory cytocompatibility, and antioxidant, anti-inflammatory, and antibacterial properties. Strikingly, in vivo, the TSNH dressing could regulate the diabetic wound microenvironment, thereby promoting collagen deposition, facilitating angiogenesis, and reducing the inflammatory response, which promotes the rapid healing of chronic wounds. This study highlights the potential of synergizing mechanical and biochemical signals in enhancing chronic wound treatment. Overall, this TSNH composite dressing is provided as a reliable approach to solving the long-standing problem of chronically infected wound healing.

Promotes M1-polarization and diabetic wound healing using Prussian blue nanozymes

Long-term inflammation and impaired angiogenesis are the main reasons for the difficulty of diabetic wound healing. What to do to effectively promote vascular endothelial cell response and immune cell reprogramming is the key to diabetic skin healing. However, contemporary therapies cannot simultaneously coordinate the promotion of vascular endothelial cells and macrophage polarization, which leads to an increased rate of disability in patients with chronic diabetes. Therefore, we developed a method of repair composed of self-assembling Prussian blue nanoenzymes, which achieved synergistic support for the immune microenvironment, and also contributed to macrophage polarization in the tissue regeneration cycle, and enhanced vascular endothelial cell activity. The template hydrothermal synthesis PB-Zr nanoplatform was prepared and locally applied to wounds to accelerate wound healing through the synergistic effect of reactive oxygen species (ROS). PB-Zr significantly normalized the wound microenvironment, thereby inhibiting ROS production and inflammatory response, which may be because it inhibited the M1 polarization of macrophages in a rat model of wound. PB-Zr treatment significantly promoted the activity of vascular endothelial cells, which better promoted the growth and regeneration of other tissues in the body. The results confirmed the disease microenvironment of PB-Zr-mediated wound therapy and indicated its application in other inflammation-related diseases.

Fabrication of l-proline enriched alginate dialdehyde-gelatin hydrogel thin films for efficient wound healing applications.

Hydrogel-based wound management systems represent a promising avenue in tissue engineering for restoring and preserving the normal functionality of damaged tissues. Incorporating active components into hydrogel matrices enhances their suitability for biomedical applications. In this study, we investigated the integration of l-proline, a nonessential imino acid with largely unexplored roles in living systems, into alginate dialdehyde-gelatin hydrogel for wound healing purposes. Physicochemical properties of the resulting hydrogel film, termed ADAGLP, were meticulously evaluated, including wound healing efficacy in vitro and anti-biofilm activity against Gram-positive and Gram-negative microorganisms. Fourier-transform infrared spectroscopy (FTIR) analysis provided insights into the interaction between l-proline and ADAG. Films incorporating 0.5% l-proline were selected for comprehensive investigation. Comparative analysis revealed prolonged gelation time and increased water holding capacity of ADAGLP compared to ADAG films. Moreover, ADAGLP exhibited a significantly higher degradation rate (69.5 ± 3.2%) compared to ADAG (35.2 ± 1.6%). Remarkably, ADAGLP demonstrated cyto-compatibility, non-toxicity, and facilitated migration to the scratch area in vitro conditions. Notably, it exhibited potent anti-biofilm properties. Our findings suggest that ADAGLP hydrogel holds promise as a biomaterial for wound care, offering prolonged drug delivery and maintaining optimal moisture levels in wound areas. The incorporation of l-proline in the wound microenvironment may contribute to enhanced tissue remodeling, by inhibiting biofilm formation, further highlighting the potential of this hydrogel system in wound healing applications.