Hair Follicle Formation Research Articles

Multifunctional alginate/polydeoxyribonucleotide hydrogels for promoting diabetic wound healing

A multifunctional alginate/PDRN hydrogel system by ionic crosslinking and the Schiff base reaction between oxidized alginate (OA) and PDRN was developed in the present study. Biocompatibility assessment of the PDRN-loaded OA hydrogels showed a significant enhancement in cell viability in human dermal fibroblast (HDF) cells. In addition, hydrogels showed migratory, anti-inflammatory, intracellular reactive oxygen species scavenging, and anti-apoptotic activities. In vivo studies using a streptozotocin-induced diabetic Wister rat model indicated that OA-4PDRN had the highest percentage of wound closure (96.1 ± 2.6 %) at day 14 compared to the control (79.0 ± 2.3 %) group. This was accompanied by up-regulation of vascular endothelial growth factor (VEGF), interleukin-10 (IL-10), and transforming growth factor-beta (TGF-β) accompanied by down-regulation of pro-inflammatory markers (IL-6, IL-1β). Following histopathological observations, PDRN-loaded OA hydrogel ensured tissue safety and induced wound healing with granular tissue formation, collagen deposition, re-epithelialization, and regeneration of blood vessels and hair follicles. The downregulation of inflammatory cytokines (CD68) and expression of angiogenesis-related cytokines (CD31) in wound sites revealed the suppression of inflammation and increased angiogenesis, ensuring skin tissue regeneration in diabetic wound healing. In conclusion, the findings suggest that PDRN-loaded OA hydrogel has enormous therapeutic potential as a diabetic wound dressing.

Mesenchyme governs hair follicle induction.

Tissue interactions are essential for guiding organ development and regeneration. Hair follicle formation relies on inductive signalling between two tissues, the embryonic surface epithelium and the adjacent mesenchyme. Although previous research has highlighted the hair-inducing potential of the mesenchymal component of the hair follicle - the dermal papilla and its precursor, the dermal condensate - the source and nature of the primary inductive signal before dermal condensate formation have remained elusive. Here, we performed epithelial-mesenchymal tissue recombination experiments using hair-forming back skin and glabrous plantar skin from mouse embryos to unveil that the back skin mesenchyme is inductive even before dermal condensate formation. Moreover, the naïve, unpatterned mesenchyme was sufficient to trigger hair follicle formation even in the oral epithelium. Building on previous knowledge, we explored the hair-inductive ability of the Wnt agonist R-spondin 1 and a Bmp receptor inhibitor in embryonic skin explants. Although R-spondin 1 instigated precocious placode-specific transcriptional responses, it was insufficient for hair follicle induction, either alone or in combination with Bmp receptor inhibition. Our findings pave the way for identifying the hair follicle-inducing cue.

A Chitosan-Based Biomaterial Combined with Mesenchymal Stem Cell-Conditioned Medium for Wound Healing and Skin Regeneration.

The aim of this study was to provide a beneficial treatment effect of novel chitosan bio-polymeric material enriched with mesenchymal stem cell products derived from the canine adipose tissue (AT-MSC) on the artificial skin defect in a rabbit model. For the objectivity of the regeneration evaluation, we used histological analysis and a scoring system created by us, taking into account all the attributes of regeneration, such as inflammatory reaction, necrosis, granulation, formation of individual skin layers and hair follicles. We observed an acceleration and improvement in the healing of an artificially created skin defect after eight and ten weeks in comparison with negative control (spontaneous healing without biomaterial). Moreover, we were able to described hair follicles and epidermis layer in histological skin samples treated with a chitosan-based biomaterial on the eighth week after grafting.

MiR-133a-3p and miR-145-5p co-promote goat hair follicle stem cell differentiation by regulating NANOG and SOX9 expression.

Hair follicle stem cells (HFSCs) differentiation is a critical physiological progress in skin hair follicle (HF) formation. Goat HFSCs differentiation is one of the essential processes of superior-quality brush hair (SQBH) synthesis. However, knowledge regarding the functions and roles of miR-133a-3p and miR-145-5p in differentiated goat HFSCs is limited. To examine the significance of chi-miR-133a-3p and chi-miR-145-5p in differentiated HFSCs, overexpression and knockdown experiments of miR-133a-3p and miR-145-5p (Mimics and Inhibitors) separately or combined were performed. NANOG, SOX9, and stem cell differentiated markers (β-catenin, C-myc, Keratin 6 [KRT6]) expression levels were detected and analyzed by using real-time quantitative polymerase chain reaction, western blotting, and immunofluorescence assays in differentiated goat HFSCs. miR-133a-3p and miR-145-5p inhibit NANOG (a gene recognized in keeping and maintaining the totipotency of embryonic stem cells) expression and promote SOX9 (an important stem cell transcription factor) expression in differentiated stem cells. Functional studies showed that miR-133a-3p and miR-145-5p individually or together overexpression can facilitate goat HFSCs differentiation, whereas suppressing miR-133a-3p and miR-145-5p or both inhibiting can inhibit goat HFSCs differentiation. These findings could more completely explain the modulatory function of miR-133a-3p and miR-145-5p in goat HFSCs growth, which also provide more understandings for further investigating goat hair follicle development.

Development and characterization of polydeoxyribonucleotide (PDRN) loaded chitosan polyplex: In vitro and in vivo evaluation of wound healing activity

Polydeoxyribonucleotide (PDRN) is an accelerated diabetic wound healing therapy with promising abilities to promote cell growth, angiogenesis, collagen synthesis, and reduce inflammation where its sustainable delivery and release behavior is critical to ensure effective wound healing properties. Therefore, a nanopolyplex was developed here, by encapsulating PDRN with chitosan to affirm its delivery systematically. The physicochemical characterization revealed its successful encapsulation which facilitates the gradual release of PDRN. In vitro studies of the polyplex demonstrated no cytotoxicity and enhanced cell proliferation and migration properties with high antimicrobial activities. In vivo, wound healing studies in Wistar rats dorsal skin defect model induced with diabetes mellitus affirm the highest wound healing activity and wound closure rate by chitosan/PDRN polyplex treatment. Considerably high histopathological changes such as epithelialization, collagen deposition, blood vessels, and hair follicle formation were observed under the polyplex treatment. The immunohistochemical analysis for platelet endothelial cell adhesion molecule (CD31) and cluster of differentiation (CD68) revealed the ability of polyplex to increase CD31 expression and decrease CD68 expression thereby promoting the wound healing process. Collectively, these results suggest that significantly accelerated, high-quality wound healing effects could be obtained by the developed chitosan/PDRN polyplex and thus it could be introduced as a potential therapeutic product for diabetic wound healing.

Autophagy critically controls skin inflammation and apoptosis-induced stem cell activation

ABSTRACT Macroautophagy/autophagy is a cellular recycling program regulating cell survival and controlling inflammatory responses in a context-dependent manner. Here, we demonstrate that keratinocyte-selective ablation of Atg16l1, an essential autophagy mediator, results in exacerbated inflammatory and neoplastic skin responses. In addition, mice lacking keratinocyte autophagy exhibit precocious onset of hair follicle growth, indicating altered activation kinetics of hair follicle stem cells (HFSCs). These HFSCs also exhibit expanded potencies in an autophagy-deficient context as shown by de novo hair follicle formation and improved healing of abrasion wounds. ATG16L1-deficient keratinocytes are markedly sensitized to apoptosis. Compound deletion of RIPK3-dependent necroptotic and CASP8-dependent apoptotic responses or of TNFRSF1A/TNFR1 reveals that the enhanced sensitivity of autophagy-deficient keratinocytes to TNF-dependent cell death is driving altered activation of HFSCs. Together, our data demonstrate that keratinocyte autophagy dampens skin inflammation and tumorigenesis but curtails HFSC activation by restraining apoptotic responses. Abbreviations: ATG16L1: autophagy related 16 like 1; DMBA: 2,4-dimethoxybenzaldehyde; DP: dermal papilla; EpdSCs: epidermal stem cells; Gas6: growth arrest specific 6; HF: hair follicle; HFSC: hair follicle stem cell; IFE: interfollicular epidermis; KRT5: keratin 5; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; PMK: primary mouse keratinocyte; RIPK3: receptor-interacting serine-threonine kinase 3; scRNAseq: single-cell RNA-sequencing; SG: sebaceous gland; TEWL: transepidermal water loss; TPA: 12-O-tetradecanoylphorbol-13-acetate; TNF: tumor necrosis factor; TNFRSF1A/TNFR1: tumor necrosis factor receptor superfamily, member 1a; UMAP: uniform manifold approximation and projection

The Effect of Platelet Rich Plasma versus MSCs Derived Exosomes on the Healing of Burn Injury in Albino Rat: Histological Study

Abstract Background The clinical treatment of skin loss due to severe and massive burns or wounds continues to be a major problem in surgical procedures. Although different kinds of dressings and therapies have been developed, but most of them are expensive. Therefore, new strategies are needed to promote and help in wound healing and repair. Recent studies have demonstrated that exosomes help tissue repair because of high stability, non immune rejection and easy control of dosage and concentration. Platelet-rich plasma (PRP) is an autologous concentration of platelets in concentrated plasma. It has been used to promote soft and hard tissue healing. Aim to compare the effect of MSCs derived exosomes versus PRP on the promotion of healing of experimentally induced second degree partial thickness burn injury in adult male albino rats. Methods The thirty-two adult male albino rats were divided randomly into 4 groups. Group I (control). Group II (burn injury group) that will be left for spontaneous healing. Group III (burn injury treated with PRP intradermal injection). Group IV (burn injury treated with intradermal MSCs derived exosome). All the animals were sacrificed at 7th day (subgroup a) and 21th day (subgroup b). The skin biopsies will be taken and processed for light microscope examination. Results H&E stained section of untreated burn (subgroup IIa) showed coagulative necrosis in epidermis and papillary layer of the dermis. Sign of epithelial migration from the edge of the wound appeared. In Subgroup IIb the surface of the wound was covered by an eschar that converted partial thickness into complete thickness. This resulted in formation of granulation tissue in the dermis. In Subgroup IIIa showed the formation of thin layer of epidermis containing undifferentiated keratinocytes. The underlying dermis contained collagen fibers and beginning of new hair follicles formation. Some specimens in Subgroup IIIb revealed thin epidermis with incomplete differentiation of keratinocytes. Numerous hair follicles were detected in the dermis. However some specimens showed thickened epidermis with presence of rete ridges. The dermis showed few irregularly distributed collagen fibers in granulation tissue with absence of skin appendages. In exosome treated group (subgroup IVa) showed thin epidermis with well differentiated keratinocytes. The dermis contained collagen fibers with absence of skin appendages. Subgroup IVb was more or less similar to the control group. Conclusion The current study revealed that burn treated exosomes group was better than PRP treated group. In addition PRP treated group revealed hyperplastic changes in epidermis with delay in remodeling phase of burn healing.

In vitro induction of hair follicle signatures using human dermal papilla cells encapsulated in fibrin microgels.

Cellular spheroids have been described as an appropriate culture system to restore human follicle dermal papilla cells (hFDPc) intrinsic properties; however, they show a low and variable efficiency to promote complete hair follicle formation in in vivo experiments. In this work, a conscientious analysis revealed a 25% cell viability in the surface of the dermal papilla spheroid (DPS) for all culture conditions, questioning whether it is an appropriate culture system for hFDPc. To overcome this problem, we propose the use of human blood plasma for the generation of fibrin microgels (FM) with encapsulated hFDPc to restore its inductive signature, either in the presence or in the absence of blood platelets. FM showed a morphology and extracellular matrix composition similar to the native dermal papilla, including Versican and Collagen IV and increasing cell viability up to 85%. While both systems induce epidermal invaginations expressing hair-specific keratins K14, K15, K71, and K75 in in vitro skin cultures, the number of generated structures increases from 17% to 49% when DPS and FM were used, respectively. These data show the potential of our experimental setting for in vitro hair follicle neogenesis with wild adult hFDPc using FM, being a crucial step in the pursuit of human hair follicle regeneration therapies.

Hypoxia Treatment of Adipose Mesenchymal Stem Cells Promotes the Growth of Dermal Papilla Cells via HIF-1α and ERK1/2 Signaling Pathways.

Dermal papilla cells (DPCs) cultured in vitro induce hair follicle formation. Using a hypoxic microenvironment to culture adipose mesenchymal stem cells (ADSCs) can promote hair follicle growth. However, the exact molecular mechanisms underlying this process remain unclear. In this study, ADSCs and DPCs from Arbas Cashmere goats were used. A hypoxic microenvironment promoted the proliferation of ADSCs and increased the pluripotency of ADSCs. The growth factors vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and platelet-derived growth factor (PDGF) were upregulated in ADSCs in the hypoxia-conditioned medium (Hypo-cm). Hypo-cm also enhanced the ability of DPCs to induce hair follicle formation. Inhibitors of the ERK1/2 signaling pathway caused the expressions of growth factors that increased in hypoxic microenvironments to decrease; moreover, hypoxia-inducible factor-1α (HIF-1α) increased the expression levels of VEGF, bFGF, and PDGF and inhibited the expression of bone morphogenic protein 7 (BMP7). In conclusion, these findings improve the theoretical basis for the development of gene therapy drugs for the treatment of alopecia areata and hair thinning.

Hyaluronate- and gelatin-based hydrogels encapsulating doxycycline as a wound dressing for burn injury therapy

Infection is a critical challenge in burn wound therapy. Wound dressings with antibacterial and multifunctional abilities associated with rapid burn wound healing are urgently needed. Here, we developed a bioadhesive and injectable ECM-mimicking hydrogel dressing with antibacterial capacity for burn injury therapy, which is crosslinked by dynamic boronate ester bonds between modified hyaluronate and gelatin (HG). The antibiotic doxycycline (Doxy) was encapsulated in HG networks for drug delivery around the wound sites. The HG/Doxy hydrogel dressing shows biocompatibility and antibacterial activity against Gram-positive and Gram-negative bacteria. Applying to a rat model of burn wound, the HG/Doxy hydrogel significantly speeds up wound closure by reducing the inflammatory reaction. Furthermore, the HG/Doxy hydrogel accelerates the regeneration of the skin structure by promoting collagen deposition, blood vessel regeneration, and hair follicle formation, eventually shortening the healing periods of burn wounds. These findings demonstrated the clinical potential of the HG/Doxy hydrogels as a promising burn wound dressing. STATEMENT OF SIGNIFICANCE: A bioadhesive and injectable hydrogel dressing has been developed for burn injury therapy. The ECM-mimicking hyaluronate-gelatin (HG) hydrogel with antibacterial ability is crosslinked by dynamic boronate ester bonds for delivering antibiotic doxycycline (Doxy). The HG/Doxy hydrogels exhibit bioadhesive, shape-adaptive, and water retention abilities in closing the irregular-shaped wound and providing a moist environment. The HG/Doxy hydrogels significantly shorten the healing periods of burn wounds in rat models within 10~14 days and promote the regeneration of skin structure, which have high potential for clinical applications.

3D bioprinting of a gelatin-alginate hydrogel for tissue-engineered hair follicle regeneration.

Hair follicle (HF) regeneration remains challenging, principally due to the absence of a platform that can successfully generate the microenvironmental cues of hair neogenesis. Here, we demonstrate a 3D bioprinting technique based on a gelatin/alginate hydrogel (GAH) to construct a multilayer composite scaffold simulating the HF microenvironment in vivo. Fibroblasts (FBs), human umbilical vein endothelial cells (HUVECs), dermal papilla cells (DPCs), and epidermal cells (EPCs) were encapsulated in GAH (prepared from a mixture of gelatin and alginate) and respectively 3D-bioprinted into the different layers of a composite scaffold. The bioprinted scaffold with epidermis- and dermis-like structure was subsequently transplanted into full-thickness wounds in nude mice. The multilayer scaffold demonstrated suitable cytocompatibility and increased the proliferation ability of DPCs (1.2-fold; P<0.05). It also facilitated the formation of self-aggregating DPC spheroids and restored DPC genes associated with hair induction (ALP, β-catenin, and α-SMA). The dermal and epidermal cells self-assembled successfully into immature HFs in vitro. HFs were regenerated in the appropriate orientation in vivo, which can mainly be attributed to the hierarchical grid structure of the scaffold and the dot bioprinting of DPCs. Our 3D printed scaffolds provide a suitable microenvironment for DPCs to regenerate entire HFs and could make a significant contribution in the medical management of hair loss. This method may also have broader applications in skin tissue (and appendage) engineering. STATEMENT OF SIGNIFICANCE: Hair loss remains a challenging clinical problem that influences quality of life. Three-dimensional (3D) bioprinting has become a useful tool for the fabrication of tissue constructs for transplantation and other biomedical applications. In this study, we used a 3D bioprinting technique based on a gelatin/alginate hydrogel to construct a multi-layer composite scaffold with cuticular and corium layers to simulate the microenvironment of dermal papilla cells (DPCs) in the human body. This new approach permits the controllable formation of self-aggregating spheroids of DPCs in a physiologically relevant extracellular matrix and the initiation of epidermal-mesenchymal interactions, which results in HF formation in vivo. The ability to regenerate entire HFs should have a significant impact on the medical management of hair loss.

Hair Growth Effect of DN106212 in C57BL/6 Mouse and Its Network Pharmacological Mechanism of Action.

Centipeda minima (CMX) has been widely investigated using network pharmacology and clinical studies for its effects on hair growth via the JAK/STAT signaling pathway. Human hair follicle papilla cells exhibit hair regrowth through the expression of Wnt signaling-related proteins. However, the mechanism of action of CMX in animals has not been elucidated fully. This study examined the effect of induced hair loss and its side-effects on the skin, and observed the mechanism of action of an alcoholic extract of CMX (DN106212) on C57BL/6 mice. Our results showed that DN106212 was more effective in promoting hair growth than dimethyl sulfoxide in the negative control and tofacitinib (TF) in the positive control when mice were treated with DN106212 for 16 days. We confirmed that DN106212 promotes the formation of mature hair follicles through hematoxylin and eosin staining. We also found that the expression of vascular endothelial growth factor (Vegfa), insulin-like growth factor 1 (Igf1), and transforming growth factor beta 1 (Tgfb1) is related to hair growth using PCR. DN106212-treated mice had significantly higher expression of Vegfa and Igf1 than TF-treated ones, and inhibiting the expression of Tgfb1 had similar effects as TF treatment. In conclusion, we propose that DN106212 increases the expression of hair growth factors, promotes the development of hair follicles, and promotes hair growth. Although additional experiments are needed, DN106212 may serve as an experimental basis for research on natural hair growth-promoting agents.

Basic principles of hair follicle structure, morphogenesis, and regeneration.

Diseases affecting the hair follicle are common in domestic animals, but despite the importance of an intact skin barrier and a fully functional hair coat, knowledge about the detailed morphological features and the diversity of these complex mini-organs are often limited, although mandatory to evaluate skin biopsies with a history of alopecia. The factors that regulate the innate hair follicle formation and the postnatal hair cycle are still not completely understood in rodents, only rudimentarily known in humans, and are poorly understood in our companion animals. This review aims to summarize the current knowledge about hair follicle and hair shaft anatomy, the arrangement of hair follicles, hair follicle morphogenesis in the embryo, and the lifelong regeneration during the postnatal hair cycle in domestic animals. The role of follicular stem cells and the need for a multitude of interacting signaling events during hair follicle morphogenesis and regeneration is unquestioned. Because of the lack of state of the art methods that can be applied in rodents but are not feasible in companion animals, most of the information in this review is based on rodent studies. However, the few data from domestic animals that are available will be discussed, and it can be assumed that at least the principal molecular mechanisms are similar in rodents and other species.

IPSC-based approach for human hair follicle regeneration.

Hair follicles (HFs) are a multifunctional structure involved in physical protection, thermoregulation, sensational detection, and wound healing. Formation and cycling of HFs require dynamic interaction between different cell types of the follicles. Although the processes have been well studied, the generation of human functional HFs with a normal cycling pattern for clinical utilization has yet to be achieved. Recently, human pluripotent stem cells (hPSCs) serve as an unlimited cell source for generating various types of cells including cells of the HFs. In this review, HF morphogenesis and cycling, different cell sources used for HF regeneration, and potential strategies for HF bioengineering using induced pluripotent stem cells (iPSCs) are depicted. Challenges and perspectives toward the therapeutic use of bioengineered HFs for hair loss disorder are also discussed.

17. Mechanical Offloading Prevents and Rescues Microtrauma Induced Fibrosis in Large Animal Model

PURPOSE: Repetitive subdermal injections, as seen with insulin in diabetes mellitus, cause microtrauma leading to skin fibrosis and consequential pain. We hypothesized that microtrauma induced fibrosis could be attenuated by injecting through a mechanical offloading dressing (MOD). We developed a large animal porcine model to evaluate injection associated fibrosis. METHODS: Two, Red-Duroc (3-month-old) pigs were subjected to four treatment conditions over a 16-week period, in two 8-week phases. All conditions received daily bleomycin injections in Phase 1 to induce microtrauma induced fibrosis. Group 1 (NN) received no MOD. Group 2 (EE) received the MOD through Phases 1, 2. Group 3 (EN) received the MOD in Phase 1 only. Lastly group 4 (NE) received the MOD in Phase 2 only. Biopsies from each group, with their respective adjacent unwounded skin (UW), were acquired at the 16-week timepoint for both histology and transcriptional analysis using single-cell-RNA sequencing. RESULTS: Skin histology by H&E and Masson trichome demonstrated that EE-treated skin significantly reduced dermal thickness and restored hair follicle formation (*p<0.05). Picrosirius red staining confirmed that EE-treatment resembled the architecture of unwounded skin (*p<0.05). Mechanical testing of EE, EN, and NE treated skin exhibited significantly decreased stiffness compared to NN skin (*p<0.05). Finally, skin deformation, was significantly greater for EE, EN, and NE treated skin (*p<0.05). scRNA-seq analysis identified specific profibrotic notch pathways responsible for fibrosis in NN treated skin, which were downregulated following MOD application. CONCLUSION: To date there is no effective treatment to prevent injection induced fibrosis, which is a huge clinical problem in diseases such as diabetes. This pig study demonstrates for the first time that mechanical offloading may prevent and treat microtrauma induced fibrosis.

Injectable chitosan/xyloglucan composite hydrogel with mechanical adaptivity and endogenous bioactivity for skin repair

Delayed or chronic wound healing is one of severe clinical issues. Developing scaffold materials capable of supporting cells and inducing tissue regeneration remains a challenge. Here, a polysaccharide-based hydrogel is constructed for promoting full-thickness skin wound healing in mouse model. The engineering hydrogel consists of a dynamic crosslinking network formed by the Schiff base reaction between aldehyde-containing xyloglucan and methacrylated chitosan. Its reversible gel-sol-gel transition upon shearing force is highly beneficial to completely cover and fill irregular wound shape. The second covalent cross-linking network achieved by photo-initiated polymerization offers a feasible way to tune the mechanical property of hydrogel after injection, with an ideal mechanical adaptivity for clinical application. Remarkably, both in vitro and in vivo evaluations demonstrate that the hydrogel with endogenously bioactive galactoside units can promote cell spheroid formation and accelerate wound healing by expediting re-epithelialization, collagen deposition, angiogenesis as well as the formation of hair follicles.

Melatonin promotes the development of the secondary hair follicles by regulating circMPP5

BackgroundThe quality and yield of cashmere fibre are closely related to the differentiation and development of secondary hair follicles in the skin of cashmere goats. The higher the density of secondary hair follicles, the higher the quality and yield of cashmere from the fleece. Development of secondary hair follicles commences in the embryonic stage of life and is completed 6 months after birth. Preliminary experimental results from our laboratory showed that melatonin (MT) treatment of goat kids after their birth could increase the density of secondary hair follicles and, thus, improve the subsequent yield and quality of cashmere. These changes in the secondary hair follicles resulted from increases in levels of antioxidant and expression of anti-apoptotic protein, and from a reduction in apoptosis. The present study was conducted to explore the molecular mechanism of MT-induced secondary hair follicle differentiation and development by using whole-genome analysis.ResultsMT had no adverse effect on the growth performance of cashmere kids but significantly improved the character of the secondary hair follicles and the quality of cashmere, and this dominant effect continued to the second year. Melatonin promotes the proliferation of secondary hair follicle cells at an early age. The formation of secondary hair follicles in the MT group was earlier than that in the control group in the second year. The genome-wide data results involved KEGG analysis of 1044 DEmRNAs, 91 DElncRNAs, 1054 DEcircRNAs, and 61 DEmiRNAs which revealed that the mitogen-activated protein kinase (MAPK) signaling pathway is involved in the development of secondary hair follicles, with key genes (FGF2, FGF21, FGFR3, MAPK3 (ERK1)) being up-regulated and expressed. We also found that the circMPP5 could sponged miR-211 and regulate the expression of MAPK3.ConclusionsWe conclude that MT achieves its effects by regulating the MAPK pathway through the circMPP5 sponged the miR-211, regulating the expression of MAPK3, to induce the differentiation and proliferation of secondary hair follicle cells.In addition there is up-regulation of expression of the anti-apoptotic protein causing reduced apoptosis of hair follicle cells. Collectively, these events increase the numbers of secondary hair follicles, thus improving the production of cashmere from these goats.

83 Wnt signaling attenuates mechanotransduction and protects against wound occlusion-mediated abolishment of regeneration

OBJECTIVES/GOALS: Current clinical practice recommends occlusive dressings (e.g., film and hydrocolloid) for wounds with variable regenerative capacities. However, clinical evidence suggests that occlusion may hinder regeneration. Our objective was to test the impact of occlusion on regeneration using animal models. METHODS/STUDY POPULATION: The murine wound-induced hair neogenesis (WIHN) is a well-established model of regeneration characterized by de novo hair follicle (HF) formation in the center of large full-thickness wounds. The quantity of neogenic HFs depends on the robustness of Wnt signaling. Optimal tissue mechanics is also required for WIHN. Utilizing the murine WIHN model, we tested the hypothesis that wound occlusion impedes regeneration. We determined how (i) the timing and duration of wound occlusion impacts WIHN, (ii) Wnt signaling influences the occlusion-induced effects on regeneration and (iii) occlusion alters the tissue mechanics of the wound, which establishes the morphogenetic field needed for WIHN. RESULTS/ANTICIPATED RESULTS: Occlusion completely eliminated WIHN. Only a brief period of occlusion between post-wound days 0-3 or 4-7 was sufficient to abrogate WIHN. Microarray and qPCR of open and occluded wounds demonstrated that occlusion promotes fibrosis by upregulating TGF-β2 and mechanotransduction, a mechanosensitive profibrotic pathway. Recruitment of these potent profibrotic pathways generated a symmetrically rigid wound incapable of de novo HF regeneration. Using transgenic animal models with enhanced Wnt signaling, we determined that the ligand-dependent Wnt signaling protected against the occlusion-induced inhibition of WIHN, as well as the occlusion-induced upregulation of both profibrotic pathways. DISCUSSION/SIGNIFICANCE: In animal models, occlusion promoted fibrosis at the expense of regeneration during acute wound healing. Augmented Wnt signaling protected against this effect. Occluding wounds may reduce regeneration. Further studies are warranted to validate these findings.

A bio-polymeric scaffold incorporated with p-Coumaric acid enhances diabetic wound healing by modulating MMP-9 and TGF-β3 expression

Diabetic wounds lead to severe health complications as the tissue regeneration process fails predominantly due to prolonged inflammation, reactive oxygen species generation, and imbalance in collagen turnover. Modern wound dressings that can aid in wound management thus improving the public healthcare system, is the present need. This study aims to fabricate an effective wound dressing using plant polyphenol to treat chronic wounds as polyphenols possess excellent wound-healing ability. The collagen scaffold enriched with the polyphenol, p-Coumaric acid, was fabricated by freeze-drying method (Col-OxP3-Ca) and examined for its wound-healing ability by in vitro and in vivo analyses. Col-OxP3-Ca scaffold exhibited 85% antioxidant activity, biocompatibility in fibroblast cells, enhanced cell proliferation and migration rate. The diabetic excision wound treated with Col-OxP3-Ca scaffold healed within 21 days and a well-developed epidermis, blood vessels, hair follicle formation, fewer inflammatory cells and collagen deposition was observed in histological analysis. The immunohistochemical results depicted the enhanced expression of TGF-β3 and lessened expression of the MMP-9 in Col-OxP3-Ca scaffold treatment group. p-Coumaric acid shortened the inflammatory stage, enhanced angiogenesis, tissue regeneration and balanced collagen turnover during healing. From this, we can accomplish that the Col-OxP3-Ca wound dressing could be an outstanding alternative to treat chronic wounds.

Platelet-rich plasma-loaded bioactive chitosan@sodium alginate@gelatin shell-core fibrous hydrogels with enhanced sustained release of growth factors for diabetic foot ulcer healing

The ability of autologous platelet-rich plasma (PRP) gel to promote rapid wound healing without immunological rejection has opened new avenues for the treatment of diabetic foot wounds. However, PRP gel still suffers from the quick release of growth factors (GFs) and requires frequent administration, thus resulting in decreased wound healing efficiency, higher cost as well as greater pain and suffering for the patients. In this study, the flow-assisted dynamic physical cross-linked coaxial microfluidic three-dimensional (3D) bio-printing technology, combined with the calcium ion chemical dual cross-linking method was developed to design PRP-loaded bioactive multi-layer shell-core fibrous hydrogels. The prepared hydrogels exhibited outstanding water absorption-retention capacity, good biocompatibility as well as a broad-spectrum antibacterial effect. Compared with clinical PRP gel, these bioactive fibrous hydrogels displayed a sustained release of GFs, reducing the administration frequency by 33 % availably during the wound treatment, but more prominent therapeutic effects such as effective reduced inflammation, in addition to promoting the growth of granulation tissue and angiogenesis, the formation of high-density hair follicles, and the generation of regular ordered and high-density collagen fiber network, which suggested great promise as exceptional candidates for treatment of diabetic foot ulcer in clinical settings.