Full-thickness Skin Defect Model Research Articles

Superior hemostatic and wound-healing properties of tetrastigma polysaccharide

Hemostatic materials that allow fast and convenient hemostasis are urgently needed. However, obtaining an ideal hemostatic agent that exhibits biocompatibility, biodegradability, and hemostatic efficacy remains challenging. Recent studies demonstrate that morphological characteristics of hemostatic materials impact their efficacy. This study reports on the importance of morphological modification of traditional Chinese medicine plant polysaccharides. The hemostatic properties of Three-leaf Qing polysaccharides, optimized in particle (SYQ-P) and sponge (SYQ-S) forms, were investigated in vitro and in vivo. SYQ-S and SYQ-P exhibit excellent biodegradability and biocompatibility, thereby improving fibroblast cell proliferation. The hemostatic mechanism involves stable blood clot formation through intrinsic coagulation pathways. In a lethal mouse liver defect bleeding model, blood loss and clotting time were lower with polysaccharides use than with commercial hemostatic drugs. SYQ-S demonstrates instant water-triggered expansion and high absorption capacity, while the sponge-like morphological characteristics effectively improve blood component concentrations. With efficient hemostasis and excellent biocompatibility, SYQ-S dramatically accelerated wound healing in a full-thickness skin defect model in vivo. Thus, SYQ-S effectively promotes the formation of granulation tissue, collagen deposition, and angiogenesis, which ultimately accelerates wound healing. Our work highlights the design of natural Chinese medicine polysaccharides and presents a promising hemostatic agent for widespread clinical application.

Photodynamic-Controllable microneedle composite with Antibacterial, Antioxidant, and angiogenic effects to expedite infected diabetic wound healing

Effective treatment of infected diabetic wound (IDW) requires delivering therapeutic agents to both wound surface and deep layers, as well as improving the wound microenvironment through antibacterial actions, clearing excessive reactive oxygen species (ROS), and promoting vascular regeneration. Here, we present a novel inorganic nanosheet called MnO2/PDA@Cu, consisting of manganese dioxide (MnO2) with copper (Cu) ions loaded on its surface through in-situ polymerization, using polydopamine (PDA) as an intermediate. We then encapsulated the MnO2/PDA@Cu nanosheet into a methacrylated hyaluronic acid (HA) soluble microneedle patch, creating a photodynamic-controllable multifunctional MnO2/PDA@Cu-HA microneedle. We used this microneedle to treat IDWs in a rat full-thickness skin defect model. Results demonstrated that the MnO2/PDA@Cu-HA microneedle effectively delivered therapeutic agents to both wound surface and deep layers, where PDA's photodynamic therapy swiftly combated bacteria under 808 nm near-infrared irradiation. Simultaneously, MnO2 acted as a nanoenzyme, mitigating ROS levels and providing oxygen for wound healing. In addition, the photothermal effect of PDA further facilitated the release of Cu, promoting vascularization. The MnO2/PDA@Cu-HA microneedle significantly accelerated wound closure, re-epithelialization, and collagen deposition, enhancing IDW healing. In conclusion, our synthesized MnO2/PDA@Cu-HA microneedle represents a photodynamic-controllable multifunctional platform for IDW treatment, offering a promising approach for effective wound management.

Synergistic Antimicrobial Glycyrrhizic Acid-based Functional Biosensing composite for Sensitive Glucose Monitoring and Collaborative Wound Healing.

High glucose blood and bacterial infection remain major issues for the slow healing of diabetic wounds, so developing functional biosensing composite with excellent antibacterial and remarkable glucose response sensitivity is necessary and prospective. Herein, by in situ synthesis AgNPs on the surface of self-prepared PTIGA elastomers, PTIGA-AgNPs conductive composites were obtained with efficient synergistic antibacterial effect, excellent mechanical, and self-healing properties. The strain of the composites could reach 1800%, and its self-healing efficiency exceeded 90% at 60°C within 8 hours. Both elastomers and composites represented excellent biocompatibility and the antibacterial rate against E coli and S aureus exceeded 90%. Moreover, the biosensor assembled from the conductive composites exhibited excellent glucose response sensitivity and stability, with a sensitivity coefficient (SC) of 0.518mA mM-1 in the range of 0.2-3.6mM glucose concentration, as well as a low detection limit (DL) of 0.08mM. Furthermore, based on the remarkable antibacterial performance and bioactivity derived from GA, the composites reduced the expression of pro-inflammatory factors and promoted the production of anti-inflammatory factors, and effectively promoted the regeneration of skin and granulation tissue of wounds in a diabetic full-thickness skin defect model, demonstrating the enormous therapeutic potential in diabetic wound healing. This article is protected by copyright. All rights reserved.

Fabrication of Quercetin-Functionalized Morpholine and Pyridine Motifs-Laden Silk Fibroin Nanofibers for Effective Wound Healing in Preclinical Study.

Choosing suitable wound dressings is crucial for effective wound healing. Spun scaffolds with bioactive molecule functionalization are gaining attention as a promising approach to expedite tissue repair and regeneration. Here, we present the synthesis of novel multifunctional quercetin with morpholine and pyridine functional motifs (QFM) embedded in silk fibroin (SF)-spun fibers (SF-QFM) for preclinical skin repair therapies. The verification of the novel QFM structural arrangement was characterized using ATR-FTIR, NMR, and ESI-MS spectroscopy analysis. Extensive characterization of the spun SF-QFM fibrous mats revealed their excellent antibacterial and antioxidant properties, biocompatibility, biodegradability, and remarkable mechanical and controlled drug release capabilities. SF-QFM mats were studied for drug release in pH 7.4 PBS over 72 h. The QFM-controlled release is mainly driven by diffusion and follows Fickian's law. Significant QFM release (40%) occurred within the first 6 h, with a total release of 79% at the end of 72 h, which is considered beneficial in effectively reducing bacterial load and helping expedite the healing process. Interestingly, the SF-QFM-spun mat demonstrated significantly improved NIH 3T3 cell proliferation and migration compared to the pure SF mat, as evidenced by the complete migration of NIH 3T3 cells within 24 h in the scratch assay. Furthermore, the in vivo outcome of SF-QFM was demonstrated by the regeneration of fresh fibroblasts and the realignment of collagen fibers deposition at 9 days post-operation in a preclinical rat full-thickness skin defect model. Our findings collectively indicate that the SF-QFM electrospun nanofiber scaffolds hold significant capability as a cost-effective and efficient bioactive spun architecture for use in wound healing applications.

Dehydration-Toughing Dual-Solvent Gels with Viscoelastic Transition for Infectious Wound Treatment.

The modulus of traditional biomedical hydrogels increases exponentially meditated by dehydration-stiffing mechanism, which leads to the failure of interface matching between hydrogels and soft tissue wounds. It is found in the study that the dual-solvent gels exhibit dehydration-toughening mechanism with the slowly increasing modulus that are always match the soft tissue wounds. Therefore, dual-solvent glycerol hydrogels (GCFen-gly DGHs) are prepared with hydrophobically modified catechol chitosan (hmCSC) and gelatin based on the supramolecular interactions. GCFen-gly DGHs exhibit excellent water retention capacity with a total solvent content exceeding 80%, permanent skin-like modulus within a range of 0.45 to 4.13kPa, and stable photothermal antibacterial abilities against S, aureus, E. coli, as well as MRSA. Infectious full-thickness rat skin defect model and tissue section analysis indicate that GCFen-gly DGHs are able to accelerate infectious wound healing by alleviating the inflammatory response, promoting granulation tissue growth, re-epithelialization, collagen deposition, and vascular regeneration. As a result, GCFen-gly DGHs is expected to become the next-generation biological gel materials for infectious wound treatment.

A Spatiotemporal Controllable Biomimetic Skin for Accelerating Wound Repair.

Skin injury repair is a dynamic process involving a series of interactions over time and space. Linking human physiological processes with materials' changes poses a significant challenge. To match the wound healing process, a spatiotemporal controllable biomimetic skin is developed, which comprises a three-dimensional (3D) printed membrane as the epidermis, a cell-containing hydrogel as the dermis, and a cytokine-laden hydrogel as the hypodermis. In the initial stage of the biomimetic skin repair wound, the membrane frame aids wound closure through pre-tension, while cells proliferate within the hydrogel. Next, as the frame disintegrates over time, cells released from the hydrogel migrate along the residual membrane. Throughout the process, continuous cytokines release from the hypodermis hydrogel ensures comprehensive nourishment. The findings reveal that in the rat full-thickness skin defect model, the biomimetic skin demonstrated a wound closure rate eight times higher than the blank group, and double the collagen content, particularly in the early repair process. Consequently, it is reasonable to infer that this biomimetic skin holds promising potential to accelerate wound closure and repair. This biomimetic skin with mechanobiological effects and spatiotemporal regulation emerges as a promising option for tissue regeneration engineering.

Mussel-inspired electroactive, antibacterial and antioxidative composite membranes with incorporation of gold nanoparticles and antibacterial peptides for enhancing skin wound healing

Large skin wounds are one of the most important health problems in the world. Skin wound repair and tissue regeneration are complex processes involving many physiological signals, and effective wound healing remains an enormous clinical challenge. Therefore, there is an urgent need for a strategy to rapidly kill bacteria, promote cell proliferation and accelerate wound healing. At present, electrical stimulation (ES) is often used in the clinical treatment of skin wounds and can simulate the endogenous biological current of the body and accelerate the repair process of skin wounds. However, a single ES strategy has difficulty covering the entire wound area, which may lead to unsatisfactory therapeutic effects. To overcome this deficiency, it is essential to develop a collaborative treatment strategy that combines ES with other treatments. In this study, gold nanoparticles and antibacterial peptides (Os) were loaded on the surface of poly(lactic-co-glycolic acid) (PLGA) material through the reducibility and adhesion of polydopamine (PDA) and improved the electrical activity, anti-inflammatory, antibacterial and biocompatibility properties of the polymer material. At the same time, this composite membrane material (Os/Au-PDA@PLGA) combined with ES was used in wound therapy to improve the wound healing rate. The results show that the new wound repair material has good biocompatibility and can effectively promote cell proliferation and migration. Through the combined application of gold nanoparticles and antibacterial peptides Os, the polymer materials have more efficient bactericidal and antioxidant effects. The antibacterial experiment results showed that gold nanoparticles could further enhance the antibacterial activity of antibacterial peptides. Furthermore, the Os/Au-PDA@PLGA composite membrane has good hydrophilicity and electrical activity, which can provide a more favorable cell microenvironment for wound healing. In vivo studies using a full-thickness skin defect model in rats showed that the Os/Au-PDA@PLGA composite membrane had a better therapeutic effect than the pure PLGA material. More importantly, the combination of the Os/Au-PDA@PLGA composite with ES significantly accelerated the rate of vascularization and collagen deposition and promoted wound healing compared with non-ES controls. Therefore, the combination of the Au/Os-PDA@PLGA composite membrane with ES may provide a new strategy for the effective treatment of skin wounds.

Rapid innervation and physiological epidermal regeneration by bioengineered dermis implanted in mouse

Tissue-engineered skin substitutes are promising tools to cover large and deep skin defects. However, the lack of a synergic and fast regeneration of the vascular network, nerves, and skin appendages limits complete skin healing and impairs functional recovery. It has been highlighted that an ideal skin substitute should mimic the structure of the native tissue to enhance clinical effectiveness. Here, we produced a pre-vascularized dermis (PVD) comprised of fibroblasts embedded in their own extracellular matrix (ECM) and a capillary-like network. Upon implantation in a mouse full-thickness skin defect model, we observed a very early innervation of the graft in 2 weeks. In addition, mouse capillaries and complete epithelialization were detectable as early as 1 week after implantation and, skin appendages developed in 2 weeks. These anatomical features underlie the interaction with the skin nerves, thus providing a further cue for reinnervation guidance. Further, the graft displays mechanical properties, collagen density, and assembly features very similar to the host tissue. Taken together our data show that the pre-existing ECM components of the PVD, physiologically organized and assembled similarly to the native tissue, support a rapid regeneration of dermal tissue. Therefore, our results suggest a promising potential for PVD in skin regeneration.

Enzymatic functionalization of decellularized tilapia skin scaffolds with enhanced skin regeneration.

The decellularized tilapia skin (dTS) has gained significant attention as a promising material for tissue regeneration due to its ability to provide unique structural and functional components that support cell growth, adhesion, and proliferation. However, the clinical application of dTS is limited by its low mechanical strength and rapid biodegradability. Herein, we prepare a novel RGD (arginine-glycine-aspartic acid) functionalized dTS scaffold (dTS/RGD) by using transglutaminase (TGase) crosslinking. The developed dTS/RGD scaffold possesses excellent properties, including a medium porosity of ∼59.2%, a suitable degradation rate of approximately 80% over a period of two weeks, and appropriate mechanical strength with a maximum tensile stress of ∼46.36 MPa which is much higher than that of dTS (∼32.23 MPa). These properties make the dTS/RGD scaffold ideal for promoting cell adhesion and proliferation, thereby accelerating skin wound healing in a full-thickness skin defect model. Such an enzymatic cross-linking strategy provides a favorable microenvironment for wound healing and holds great potential for application in skin regeneration engineering.

Yüksek yağlı diyet ile beslenen ve tam kalınlıkta deri defekti modeli oluşturulan sıçanlarda laktoferrinin etkisi

Purpose: Lactoferrin is a member of the milk protein family with a broad spectrum of bioactivities and has therapeutic effects against many microbes, viruses, and other pathogens. This study aims to show the healing effect of lactoferrin against obesity-related wound healing.
 Materials and Methods: In this study, 48 male rats were grouped as the Control (C) (n =8), Full-thickness skin defect model (FT) (n =8), Bovine Lactoferrin (bLf) (200 mg/kg)(n= 8), High Fat Diet (HFD) + Bovine Lactoferrin (bLf) (n = 8), High-Fat Diet (HFD)+Full-thickness skin defect model (FT) (n=8), and HFD+Full-thickness skin defect model (FT) + Bovine Lactoferrin bLf (n=8). High-Fat Diet + bLf and High-Fat Diet + Full-thickness skin defect + bLf group rats were given a high-fat diet and Bovine Lactoferrin orally. IL-6 and VEGF biochemical parameters were examined in serum.
 Results: When the serum IL-6 protein amounts of the groups were examined, it was the highest in the HFD+FT+bLf group on the 21st day, and the lowest in the FT group, when the serum VEGF protein amounts, which were the other biochemical parameters, were evaluated, these amounts were found to be the highest in the HFD+FT+bLf group and the lowest in the FT group on the 21st day.
 Conclusion: Lactoferrin has been shown to accelerate wound healing, and the fact that lactoferrin is readily available and abundant in milk makes it an exciting treatment option for wound healing and inflammation in the future.

Bioinspired Zn-MOF doped radial porous chitosan-based sponge with antibacterial and antioxidant properties for rapid hemostasis and wound healing

Herein, a novel bioinspired radial porous zinc-based metal-organic framework (Zn-MOF) doped sodium alginate/chitosan derivatives/pullulan-based SA/PSCS/Pul/Zn-MOF (SPCP/Zn) composites sponge with excellent antioxidant and antibacterial properties was fabricated by the ice-templating method. Boric acid (BA) and Ca2+, which were respectively used as hydrogen- and ionic- bonding cross-linkers, provided strong mechanical properties for sponge matrix composed of SA, PSCS, and Pul. The obtained SPCP/Zn sponge exhibited uniform porous morphology, proper hydrophilicity, and admirable biocompatibility. In addition, the SPCP/Zn sponge achieved a sustained release of Zn2+ and gallic acid, which displayed powerful antibacterial and antioxidant activities. Importantly, the SPCP/Zn sponge exhibited shorter rapid hemostasis (20.4 ± 2.9 s) and lower blood loss (19.8 ± 4.3 mg). The SPCP/Zn sponge also showed faster wound closure ratio for the rat full-thickness skin defect model. It was revealed that SPCP/Zn sponge could significantly accelerate and enhance wound healing through downregulating inflammatory cytokines (TNF-α, IL-6) and increasing the expression of growth factors (VEGF). Due to its excellent properties, the SPCP/Zn sponge may have promising potential in wound healing applications.

Muscle-inspired lamellar chitosan sponge with photothermal antibacterial and antioxidant properties for hemostasis and accelerated bacteria infected wound healing

Wound dressings with excellent hemostasis, antibacterial, and antioxidant properties are essential for hemorrhagic infected wounds. Herein, inspired by the highly anisotropic and ordered fibrous structure of muscle tissue, we prepared chitosan/polydopamine biomimetic layered sponge dressings loaded with cerium dioxide (CeO2) nanoparticles by a directional freezing method. This composite sponge has an ordered laminate structure, good mechanical properties, excellent moisture absorption, and swelling properties. Thanks to the introduction of PDA, the composite sponge has outstanding photothermal properties and effectively kills bacteria under 808 nm near-infrared (NIR) light. In addition, the CeO2 loaded in the composite sponge also has an excellent ROS scavenging capacity, which protects cells from oxidative stress damage. The rat tail amputation and liver hemostasis experiment proved that this layered sponge had good hemostasis performance. Furthermore, the full-thickness skin defect model infected with S. aureus demonstrated that the composite sponge accelerated wound closure, promoted collagen deposition, down-regulated pro-inflammatory factors (TNF-α), and up-regulated anti-inflammatory factors (IL-10) expression, ultimately promoting wound healing. All the results suggested that this bionic sponge with a directional layered structure has a preferable therapeutic effect on the healing of infected wounds and is expected to be a more advantageous wound dressing.

Bilayer wound dressing composed of asymmetric polycaprolactone membrane and chitosan-carrageenan hydrogel incorporating storax balsam

A comprehensive approach is needed to develop multifunctional wound dressing that is simple yet efficient. In this work, Liquidambar orientalis Mill. storax loaded hydroxyethyl chitosan (HECS)-carrageenan (kC) based hydrogel (HECS-kC) and polydopamine coated asymmetric polycaprolactone membrane (PCL-DOP) were used to develop a multifunctional and modular bilayer wound dressing. Asymmetric PCL-DOP membrane was prepared by non-solvent induced phase separation (NIPS) followed by polydopamine coating and demonstrated an excellent barrier against bacteria while allowing permeability for 5.45 ppm dissolved‑oxygen and 2130 g/m2 water vapor transmission in 24 h in addition to 805 kPa tensile strength. Storax loaded HECS-kC hydrogel, on the other hand, demonstrated a pH-responsive degradation and swelling to provide necessary conditions to facilitate wound healing. The hydrogels showed stretchability above 140 %, mild adhesive strength on sheep skin and PCL-DOP membrane, while the storax incorporation enhanced antibacterial and antioxidant activity. Furthermore, rat full-thickness skin defect model showed that the developed bilayer wound dressing could significantly facilitate wound healing compared to Tegaderm™ and control groups. This study shows that the bilayered wound dressing has the potential to be used as a simple and effective wound care system.

Dynamic Microenvironment-Adaptable Hydrogel with Photothermal Performance and ROS Scavenging for Management of Diabetic Ulcer.

Persistent bacterial infections and excessive oxidative stress prevent the healing of diabetic ulcers, leading to an increased disability rate. Current treatments fail to kill bacteria while simultaneously relieving oxidative stress. Herein, a dynamic microenvironment-adaptable hydrogel (BP@CAu) with photothermal performance and reactive oxygen species scavenging is presented for diabetic ulcer healing. This hydrogel prepared using a dynamic borate-ester could respond to acidity in the infection microenvironment for a controllable drug release. An excellent photothermal conversion effect was integrated in the hydrogel, which exhibited strong antibacterial activity against Staphylococcus aureus and Pseudomonas aeruginosa. The hydrogel attenuated intracellular oxidative stress and inflammation and promoted cell migration. In a full-thickness skin defect model of diabetic rats, the BP@CAu hydrogel contributed to the fastest wound closure, with ideal reepithelialization, granulation tissue formation, and regeneration of blood vessels. Further mechanistic studies revealed that the hydrogel relieved oxidative stress and downregulated the expression of inflammatory cytokines, resulting in dramatic therapeutic effects on diabetic wounds. Therefore, this study provides a synergistic therapeutic strategy for efficient photothermal performance and reactive oxygen species scavenging in diabetic ulcers.

An injectable bioactive dressing based on platelet-rich plasma and nanoclay: Sustained release of deferoxamine to accelerate chronic wound healing.

Delayed diabetic wound healing has placed an enormous burden on society. The key factors limiting wound healing include unresolved inflammation and impaired angiogenesis. Platelet-rich plasma (PRP) gel, a popular biomaterial in the field of regeneration, has limited applications due to its non-injectable properties and rapid release and degradation of growth factors. Here, we prepared an injectable hydrogel (DPLG) based on PRP and laponite by a simple one-step mixing method. Taking advantages of the non-covalent interactions, DPLG could overcome the limitations of PRP gels, which is injectable to fill irregular injures and could serve as a local drug reservoir to achieve the sustained release of growth factors in PRP and deferoxamine (an angiogenesis promoter). DPLG has an excellent ability in accelerating wound healing by promoting macrophage polarization and angiogenesis in a full-thickness skin defect model in type I diabetic rats and normal rats. Taken together, this study may provide the ingenious and simple bioactive wound dressing with a superior ability to promote wound healing.

Biological Macromolecule Hydrogel Based on Recombinant Type I Collagen/Chitosan Scaffold to Accelerate Full-Thickness Healing of Skin Wounds.

The development of biological macromolecule hydrogel dressings with fatigue resistance, sufficient mechanical strength, and versatility in clinical treatment is critical for accelerating full-thickness healing of skin wounds. Therefore, in this study, multifunctional, biological macromolecule hydrogels based on a recombinant type I collagen/chitosan scaffold incorporated with a metal-polyphenol structure were fabricated to accelerate wound healing. The resulting biological macromolecule hydrogel possesses sufficient mechanical strength, fatigue resistance, and healing properties, including antibacterial, antioxygenic, self-healing, vascularization, hemostatic, and adhesive abilities. Chitosan and recombinant type I collagen formed the scaffold network, which was the first covalent crosslinking network of the hydrogel. The second physical crosslinking network comprised the coordination of a metal-polyphenol structure, i.e., Cu2+ with the catechol group of dopamine methacrylamide (DMA) and stacking of DMA benzene rings. Double-crosslinked networks are interspersed and intertwined in the hydrogel to reduce the mechanical strength and increase its fatigue resistance, making it more suitable for clinical applications. Moreover, the biological macromolecule hydrogel can continuously release Cu2+, which provides strong antibacterial and vascularization properties. An in vivo full-thickness skin defect model confirmed that multifunctional, biological macromolecule hydrogels based on a recombinant type I collagen/chitosan scaffold incorporated with a metal-polyphenol structure can facilitate the formation of granulation tissue and collagen deposition for a short period to promote wound healing. This study highlights that this biological macromolecule hydrogel is a promising acute wound-healing dressing for biomedical applications.

An Injectable Composite Hydrogel of Verteporfin-Bonded Carboxymethyl Chitosan and Oxidized Sodium Alginate Facilitates Scarless Full-Thickness Skin Regeneration.

Full-thickness skin defect has always been a major challenge in clinics due to fibrous hyperplasia in the repair process. Hydrogel composite dressings loaded with anti-fibrotic drugs have been considered as a promising strategy for scarless skin regeneration. In this work, a hydrogel composite (VP-CMCS-OSA) of carboxymethyl chitosan (CMCS) and oxidized sodium alginate (OSA), with loading anti-fibrotic drug verteporfin (VP), is developed based on two-step chemical reactions. Verteporfin is bonded with carboxymethyl chitosan through EDC/NHS treatment to form VP-CMCS, and then VP-CMCS is crosslinked with oxidized sodium alginate by Schiff base reaction to form VP-CMCS-OSA hydrogel. The characterization by SEM, FTIR, and UV-Vis shows the microstructure and chemical bonding of VP-CMCS-OSA. VP-CMCS-OSA hydrogel demonstrates the properties of high tissue adhesion, strong self-healing, and tensile ability. In the full-thickness skin defect model, the VP-CMCS-OSA composite hydrogels hasten wound healing due to the synergistic effects of hydrogels and verteporfin administration. The histological examination reveals the regular collagen arrangement and more skin appendages after VP-CMCS-OSA composite hydrogel treatment, indicating the full-thickness skin regeneration without potential scar formation. The outcomes suggest that the verteporfin-loaded composite hydrogel could be a potential method for scarless skin regeneration.

Enhanced wound healing and hemostasis with exosome-loaded gelatin sponges from human umbilical cord mesenchymal stem cells.

Rapid wound healing remains a pressing clinical challenge, necessitating studies to hasten this process. A promising approach involves the utilization of human umbilical cord mesenchymal stem cells (hUC-MSCs) derived exosomes. The hypothesis of this study was that these exosomes, when loaded onto a gelatin sponge, a common hemostatic material, would enhance hemostasis and accelerate wound healing. To investigate the hemostatic and wound healing efficacy of gelatin sponges loaded with hUC-MSCs-derived exosomes. Ultracentrifugation was used to extract exosomes from hUC-MSCs. Nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), and western blot techniques were used to validate the exosomes. In vitro experiments were performed using L929 cells to evaluate the cytotoxicity of the exosomes and their impact on cell growth and survival. New Zealand rabbits were used for skin irritation experiments to assess whether they caused adverse skin reactions. Hemolysis test was conducted using a 2% rabbit red blood cell suspension to detect whether they caused hemolysis. Moreover, in vivo experiments were carried out by implanting a gelatin sponge loaded with exosomes subcutaneously in Sprague-Dawley (SD) rats to perform biocompatibility tests. In addition, coagulation index test was conducted to evaluate their impact on blood coagulation. Meanwhile, SD rat liver defect hemostasis model and full-thickness skin defect model were used to study whether the gelatin sponge loaded with exosomes effectively stopped bleeding and promoted wound healing. The NTA, TEM, and western blot experimental results confirmed that exosomes were successfully isolated from hUC-MSCs. The gelatin sponge loaded with exosomes did not exhibit significant cell toxicity, skin irritation, or hemolysis, and they demonstrated good compatibility in SD rats. Additionally, the effectiveness of the gelatin sponge loaded with exosomes in hemostasis and wound healing was validated. The results of the coagulation index experiment indicated that the gelatin sponge loaded with exosomes had significantly better coagulation effect compared to the regular gelatin sponge, and they showed excellent hemostatic performance in a liver defect hemostasis model. Finally, the full-thickness skin defect healing experiment results showed significant improvement in the healing process of wounds treated with the gelatin sponge loaded with exosomes compared to other groups. Collectively, the gelatin sponge loaded with hUC-MSCs-derived exosomes is safe and efficacious for promoting hemostasis and accelerating wound healing, warranting further clinical application.

EGCG modified small intestine submucosa promotes wound healing through immunomodulation

Decellularized porcine small intestinal submucosa (SIS) has shown promising therapeutic efficacy as a functional wound dressing. Nevertheless, its limited anti-oxidative and immunomodulatory capacities have restricted its application for the treatment of complex skin wounds. Herein, epigallocatechin gallate (EGCG), a polyphenolic compound, was employed for the modification of the SIS to overcome such shortcomings. The EGCG-modified SIS (E-SIS) has shown excellent biocompatibility and improved hydrophilicity for cell adhesion. Notably, in vitro studies showed that the E-SIS could effectively alleviate oxidative stress and facilitate the M1-to-M2 phenotype transition of macrophages, thereby creating a favorable immune microenvironment for cell proliferation, migration, collagen synthesis as well as angiogenesis. A full-thickness skin defect model, combined with macrophage depletion, has further confirmed that the E-SIS could accelerate skin wound repair through immunomodulation in vivo. This suggested that the EGCG modification could provide a facile yet effective method to broaden the applications of the SIS for skin wound management.

A multi-functional double cross-linked chitosan hydrogel with tunable mechanical and antibacterial properties for skin wound dressing

Chitosan hydrogels with essential antibacterial properties and biocompatibility have great potential in tissue engineering and regeneration medicine. However, pure chitosan hydrogel could be limited by insufficient mechanical properties. In this work, we designed a multi-functional chitosan hydrogel based on the combination of chitosan methacrylate (CTSMA) and sulfhydrated chitosan (CTSSH), which is cross-linked simultaneously by free-radical polymerization reaction and Thiol-ene reaction. The CTSMA/CTSSH (CMS) hydrogels displayed superior tissue adhesive and mechanical properties when compared to pure CTSMA hydrogel. Additionally, the resulting hydrogels exhibited potent antimicrobial effects against both E. coli and S. aureus. Besides, the CMS hydrogels exhibited good biocompatibility as demonstrated by cytotoxicity and cell proliferation experiments using fibroblasts cells (L929) and adipose-derived stem cells (ADSCs). In vivo experiment, the repairing effect of hydrogels on full-thickness skin defect model in rats was studied. Histological and immunohistochemical staining results showed that CMS hydrogels promoted angiogenesis, dermal repair and epidermal regeneration. Overall, the study highlights the potential of the CMS hydrogels as a promising biomaterial in wound healing applications.