Construction of programmed time-released multifunctional hydrogel with antibacterial and anti-inflammatory properties for impaired wound healing.

The successful reprogramming of impaired wound healing presents ongoing challenges due to the impaired tissue microenvironment caused by severe bacterial infection, excessive oxidative stress, as well as the inappropriate dosage timing during different stages of the healing process. Herein, a dual-layer hydrogel with sodium alginate (SA)-loaded zinc oxide (ZnO) nanoparticles and poly(N-isopropylacrylamide) (PNIPAM)-loaded Cu5.4O ultrasmall nanozymes (named programmed time-released multifunctional hydrogel, PTMH) was designed to dynamically regulate the wound inflammatory microenvironment based on different phases of wound repairing. PTMH combated bacteria at the early phase of infection by generating reactive oxygen species through ZnO under visible-light irradiation with gradual degradation of the lower layer. Subsequently, when the upper layer was in direct contact with the wound tissue, Cu5.4O ultrasmall nanozymes were released to scavenge excessive reactive oxygen species. This neutralized a range of inflammatory factors and facilitated the transition from the inflammatory phase to the proliferative phase. Furthermore, the utilization of Cu5.4O ultrasmall nanozymes enhanced angiogenesis, thereby facilitating the delivery of oxygen and nutrients to the impaired tissue. Our experimental findings indicate that PTMHs promote the healing process of diabetic wounds with bacterial infection in mice, exhibiting notable antibacterial and anti-inflammatory properties over a specific period of time.

COX2-ATP Synthase Regulates Spine Follicle Size in Hedgehogs.

Skin evolves essential appendages with adaptive patterns that synergistically insulate the body from environmental insults. How similar appendages in different animals generate diversely-sized appendages remain elusive. Here we used hedgehog spine follicles and mouse hair follicles as models to investigate how similar follicles form in different sizes postnatally. Histology and immunostaining show that the spine follicles have a significantly greater size than the hair follicles. By RNA-sequencing analysis, we found that ATP synthases are highly expressed in hedgehog skin compared to mouse skin. Inhibition of ATP synthase resulted in smaller spine follicle formation during regeneration. We also identified that the mitochondrial gene COX2 functions upstream of ATP synthase that influences energy metabolism and cell proliferation to control the size of the spine follicles. Our study identified molecules that function differently in forming diversely-sized skin appendages across different animals, allowing them to adapt to the living environment and benefit from self-protection.

Stability and biosafety of human epidermal stem cell for wound repair: preclinical evaluation.

BACKGROUND: Cell therapy is a key technology to prevent sacrificing normal skin. Although some studies have shown the promise of human epidermal stem cells (EpiSCs), the efficacy, biosafety and quality control of EpiSC therapy have not been systematically reported. METHODS: The biosafety, stemness maintenance and wound repair of EpiSC were systematically verified by in vitro and in vivo experiments. EpiSC were prepared from the foreskin using a collagen type IV rapid adherence method. The EpiSCs were identified by flow cytometry, immunofluorescence staining and cell morphology. The well-growing passage 1 (P1) EpiSCs were used to determine the proliferation curve (counting method). EpiSC clone formation assay was performed by Giemsa staining. Nude mice were used to prepare a full-thickness skin defect wound model to detect the repair effect of EpiSCs. The biosafety of EpiSCs was double tested in vitro and in vivo. RESULTS: The results showed that the expression of specific markers and clone formation efficiency was stable when passage 1 (P1) to P8 cells were cultured, and the stemness rate of P8 cells was close to 85.1%. EpiSCs were expanded in vitro for 25 days, the number of cells reached 2.5 × 108, and the transplantable area was approximately 75% of the total body surface area (TBSA). At 45 days, the total number of cells was approximately 30 billion, and the transplantable area was approximately the size of a volleyball court. A nude mouse wound model indicated that EpiSCs could rapidly close a wound. On postinjury day 7, the wound epithelialization rate in the cell transplantation group was significantly higher than that in the NaCl group (P < 0.05). In vitro, cell senescence increased, and telomerase activity decreased in P1 to P8 EpiSCs. In vivo, there were no solid tumors or metastatic tumors after EpiSC (P8) transplantation. In addition, the quality control of cultured cells met the clinical application criteria for cell therapy. CONCLUSION: This preclinical study showed the stability and biosafety of human EpiSC therapy for wound repair.

Nanomaterials and nanomaterials-based drug delivery to promote cutaneous wound healing.

Various factors could damage the structure and integrity of skin to cause wounds. Nonhealing or chronic wounds seriously affect the well-being of patients and bring heavy burdens to the society. The past few decades have witnessed application of numerous nanomaterials to promote wound healing. Owing to the unique physicochemical characteristics at nanoscale, nanomaterials-based therapy has been regarded as a potential approach to promote wound healing. In this review, we first overview the wound categories, wound healing process and critical influencing factors. Then applications of nanomaterials with intrinsic therapeutic effect and nanomaterials-based drug delivery systems to promote wound healing are addressed in detail. Finally, current limitations and future perspectives of nanomaterials in wound healing are discussed.

Snake extract-laden hemostatic bioadhesive gel cross-linked by visible light.

Bioadhesives reduce operation time and surgical complications. However, in the presence of blood, adhesion strength is often compromised. Inspired by the blood clotting activity of snake venom, we report a visible light-induced blood-resistant hemostatic adhesive (HAD) containing gelatin methacryloyl and reptilase, which is a hemocoagulase (HC) extracted from Bothrops atrox HAD leads to the activation and aggregation of platelets and efficiently transforms fibrinogen into fibrin to achieve rapid hemostasis and seal the tissue. Blood clotting time with HAD was about 45 s compared with 5 to 6 min without HAD. HAD instantaneously achieved hemostasis on liver incision (~45 s) and cut rat tail (~34 s) and reduced blood loss by 79 and 78%, respectively. HAD is also efficient in sealing severely injured liver and abdominal aorta. HAD has great potential to bridge injured tissues by combing hemostasis with adhesives.

A moisture balanced antibacterial dressing loaded with lysozyme possesses antibacterial activity and promotes wound healing.

Wound moisture management is very important in wound healing. Previous wound management has included dry healing and moist healing, and the theory of wound moisture balance is currently generally accepted. However, current studies have not reported which humidity is suitable for wound healing and how to appropriately use antibacterial compounds when the humidity is suitable. Our study explored the moisture balance of polyurethane foam dressings through a moisture balance test and constructed a safe and effective moisture balanced antibacterial dressing by loading lysozyme onto a polyurethane foam dressing. Wound healing experiments showed that the wound healing speed was the fastest when the humidity was 25%. In vivo and in vitro antibacterial experiments showed the superior antibacterial performance of the dressing after lysozyme loading. We loaded lysozyme on moisture balanced polyurethane dressings by means of dopamine adsorption, and the modified dressings were named PU/DA-LYS (polyurethane/dopamine-lysozyme). Experiments on wound healing in infected mice indicated that PU/DA-LYS helps fight infection while promoting wound healing. Cytotoxicity experiments and in vivo biological safety experiments indicated that PU/DA-LYS was safe for use. Our study found that the lysozyme loaded polyurethane dressing can provide appropriate wound moisture and prevent bacterial infection, which is a future developmental direction for wound dressings.

GDF-5 promotes epidermal stem cells proliferation via Foxg1-cyclin D1 signaling.

OBJECTIVE: Epidermal stem cells (EpSCs) can self-renew, which are responsible for the long-term maintenance of the skin, and it also plays a critical role in wound re-epithelization, but the mechanism underlying EpSCs proliferation is unclear. GDF-5, also known as BMP-14, is a member of the BMP family and can be used as a self-renewal supporter. Here, we studied the effects of GDF-5 on mouse EpSCs proliferation mechanism in wound healing. METHODS: Firstly, the effects of GDF-5 on EpSCs proliferation was tested by using CCK8 reagent and PCNA expression was analyzed by Western blotting. Secondly, we screened genes that promote EpSCs proliferation in the FOX and cyclin family by qPCR, and then the protein expression level of the selected genes was further analyzed by Western blotting. Thirdly, siRNA plasmids and pAdEasy adenovirus were transfected or infected, respectively, into mouse EpSCs to detect the effect of target genes on GDF-5-induced cell proliferation. Furthermore, we injected GDF-5 to a deep partial thickness burn mouse model for finding out whether EpSCs proliferation can be detected by immunohistochemical. Finally, the relevant target genes were analyzed by qPCR, immunoblotting, and dual-luciferase reporter gene detection. RESULTS: We discovered that 100 ng/ml recombinant mouse GDF-5 was the optimal concentration for promoting mouse EpSCs proliferation. Through preliminary screened by qPCR, we found that Foxg1 and cyclin D1 could be the downstream molecules of GDF-5, and the results were confirmed by Western blotting. And the effect of GDF-5 on mouse EpSCs proliferation was adjusted by Foxg1/cyclin D1 in vitro and in vivo. Besides, GDF-5-induced transcription of cyclin D1 was regulated by Foxg1-mediated cyclin D1 promoter activity. CONCLUSION: This paper showed that GDF-5 promotes mouse EpSCs proliferation via Foxg1-cyclin D1 signal pathway. It is suggested that GDF-5 may be a new approach to make EpSCs proliferation which can be used in wound healing.

GDF-5 induces epidermal stem cell migration via RhoA-MMP9 signalling.

The migration of epidermal stem cells (EpSCs) is critical for wound re-epithelization and wound healing. Recently, growth/differentiation factor-5 (GDF-5) was discovered to have multiple biological effects on wound healing; however, its role in EpSCs remains unclear. In this work, recombinant mouse GDF-5 (rmGDF-5) was found via live imaging in vitro to facilitate the migration of mouse EpSCs in a wound-scratch model. Western blot and real-time PCR assays demonstrated that the expression levels of RhoA and matrix metalloproteinase-9 (MMP9) were correlated with rmGDF-5 concentration. Furthermore, we found that rmGDF-5 stimulated mouse EpSC migration in vitro by regulating MMP9 expression at the mRNA and protein levels through the RhoA signalling pathway. Moreover, in a deep partial-thickness scald mouse model in vivo, GDF-5 was confirmed to promote EpSC migration and MMP9 expression via RhoA, as evidenced by the tracking of cells labelled with 5-bromo-2-deoxyuridine (BrdU). The current study showed that rmGDF-5 can promote mouse EpSC migration in vitro and in vivo and that GDF-5 can trigger the migration of EpSCs via RhoA-MMP9 signalling.

Epidemiology and Outcome Analysis of 470 Patients with Hand Burns: A Five-Year Retrospective Study in a Major Burn Center in Southwest China.

BACKGROUND This retrospective study aimed to investigate the epidemiology of burns to the hand, including the causes, demographic data, management, and outcome in a single center in Southwest China between 2012 and 2017. MATERIAL AND METHODS A retrospective study included 470 patients with hand burns who were treated at a single hospital in Southwest China between 2012 and 2017. Demographic, injury-related, and clinical data were obtained from the clinical electronic data collection system. RESULTS In 470 patients, men were more commonly admitted to hospital with hand burns (73.62%). Children under 10 years (29.57%) were the main patient group. Hospital admissions occurred in the coldest months, from December to March (55.11%). In 60.21% of cases, hand burns occurred outside the workplace. Fire (40.42%), electricity (30.85%), and hot liquids (20.21%) were the main causes of hand burns. Data from 428 patients showed that burns with a larger total body surface area and deeper burns were associated with surgery and amputation. Burn depth was a risk factor for skin grafting, and lack of burn cooling before hospital admission increased the risk of amputation. Data from 117 patients with localized burns showed that full-thickness burns and lack of cooling before admission were associated with an increased hospital stay. CONCLUSIONS The findings suggest that in Southwest China, prevention programs for children aged 0-9 years, injuries occurring in winter and non-workplace sites, and fire burns were imperative.

Epidemiological and clinical characteristics of burns in the older person: a seven-year retrospective analysis of 693 cases at a burn center in south-west China.

BACKGROUND: Burns are one of the major traumas that may affect older individuals. The purpose of this study was to investigate the epidemiological and clinical characteristics of geriatric burns at a major center in south-west China. METHODS: This retrospective study was conducted at the Institute of Burn Research, Southwest Hospital of Army Medical University between 2010 and 2016, and the data collected from medical records included admission date, age, gender, premorbid disease, burn etiology, injured anatomical location, burn area and depth, inhalation injury, number of surgeries, length of stay (LOS), clinical outcome, and medical cost. RESULTS: Of the 693 older burn patients included, 60.75% were male and 56.85% were aged 60-69 years. Burns peaked in December-March and June. Flame was the most common cause of burns, making up 51.95% of all cases, and also dominated in the burn patients aged 60-69 years. Limbs were the most common anatomical sites of burns (69.41%), and the median total body surface area (TBSA) was 5% (interquartile range [IQR]: 2%-15%). The percentage of the patients who underwent surgeries and number of surgeries significantly increased in the cases of contact burns, younger age and full-thickness burns. Six deaths resulted in a mortality of 0.9%. The median LOS was 16 days (IQR: 8-29 days), and the main risk factors were more surgeries, better outcomes, and full-thickness burns. The median cost was 20,228 CNY (IQR: 10,457- 46,581.5 CNY), and major risk factors included longer LOS, larger TBSA, and more surgeries. Furthermore, compared to the earlier data from our center, the proportion of older adults among all burns (7.50% vs. 4.15%), proportion of flame burns (51.95% vs. 33.90%), and mean age (69.05 years vs. 65.10 years) were significantly higher, while the proportion of premorbidities (16.9% vs. 83.9%), mortality (0.9% vs. 7.5%) and median TBSA (5% vs. 21%) were significantly lower. CONCLUSIONS: This study suggested that closer attention should be paid to prevent burn injuries in older people aged 60-69 years, especially males, regarding incidents in the summer and winter, and flame burns. Moreover, tailored intervention strategies based on related risk factors should be under special consideration.

Corrigendum: Reconstruction and Functional Annotation of P311 Protein-Protein Interaction Network Reveals Its New Functions.

[This corrects the article DOI: 10.3389/fgene.2019.00109.].

NCF2, MYO1F, S1PR4, and FCN1 as potential noninvasive diagnostic biomarkers in patients with obstructive coronary artery: A weighted gene co-expression network analysis.

This study aims to explore the predictive noninvasive biomarker for obstructive coronary artery disease (CAD). By using the data set GSE90074, weighted gene co-expression network analysis (WGCNA), and protein-protein interactive network, construction of differentially expressed genes in peripheral blood mononuclear cells was conducted to identify the most significant gene clusters associated with obstructive CAD. Univariate and multivariate stepwise logistic regression analyses and receiver operating characteristic analysis were used to predicate the diagnostic accuracy of biomarker candidates in the detection of obstructive CAD. Furthermore, functional prediction of candidate gene biomarkers was further confirmed in ST-segment elevation myocardial infarction (STEMI) patients or stable CAD patients by using the datasets of GSE62646 and GSE59867. We found that the blue module discriminated by WGCNA contained 13 hub-genes that could be independent risk factors for obstructive CAD (P < .05). Among these 13 hub-genes, a four-gene signature including neutrophil cytosol factor 2 (NCF2, P = .025), myosin-If (MYO1F, P = .001), sphingosine-1-phosphate receptor 4 (S1PR4, P = .015), and ficolin-1 (FCN1, P = .012) alone or combined with two risk factors (male sex and hyperlipidemia) may represent potential diagnostic biomarkers in obstructive CAD. Furthermore, the messenger RNA levels of NCF2, MYO1F, S1PR4, and FCN1 were higher in STEMI patients than that in stable CAD patients, although S1PR4 showed no statistical difference (P > .05). This four-gene signature could also act as a prognostic biomarker to discriminate STEMI patients from stable CAD patients. These findings suggest a four-gene signature (NCF2, MYO1F, S1PR4, and FCN1) alone or combined with two risk factors (male sex and hyperlipidemia) as a promising prognostic biomarker in the diagnosis of STEMI. Well-designed cohort studies should be implemented to warrant the diagnostic value of these genes in clinical purpose.

Hematopoietic-Specific Deletion of Foxo1 Promotes NK Cell Specification and Proliferation.

We previously reported that deletion of Foxo1, via Ncr1-iCre mice from the expression of NKp46 onward, led to enhanced natural killer (NK) cell maturation and effector function. In this model, however, the role of Foxo1 in regulating NK cell specification and early development remains exclusive. Herein, we utilized a murine model of hematopoietic-specific deletion of Foxo1 before lymphoid specification, by crossing mice carrying floxed Foxo1 alleles (Foxo1fl/fl) with Vav1-iCre mice, to revisit the role of Foxo1 on NK cell specification and early development. The data showed that hematopoietic-specific deletion of Foxo1 resulted in increased proportion and numbers of common lymphoid progenitors (CLP) (Lin-CD127+c-Kit+Sca-1+), pre-pro NK b cells (Lin-Sca-1+c-Kit-CD135-CD127+), as well as committed Lin-CD122+ cells and CD3-CD19-NKp46+ NK cells in bone marrow. Hematopoietic-specific deletion of Foxo1 also promoted NK cells proliferation in a cell-intrinsic manner, indicated by increased Ki-67 expression and more expansion of NK cell after ex vivo stimulation with IL-15. The reason for Foxo1 suppressing NK cell proliferation might be its direct transcription of the cell-cycle inhibitory genes, such as p21cip1, p27kip1, p130, Gadd45a, and Ccng2 (cyclin G2) in NK cells, supported by the evidence of decreased mRNA expression of p21cip1, p27kip1, p130, Gadd45a, and Ccng2 in Foxo1-deficient NK cells and direct binding of Foxo1 on their promoter region. Furthermore, hematopoietic-specific deletion of Foxo1 resulted in increased ratio of mature NK subsets, such as CD11b+CD27- and CD43+KLRG1+ NK cells, but decreased ratio of immature NK subsets, such as CD27+CD11b- and CD27+CD11b+ NK cells, consistent with the findings in the murine model of Ncr1-iCre mediated Foxo1 deletion. Conclusively, Foxo1 not only acts as a negative checkpoint on NK cell maturation, but also represses NK cell specification and proliferation. The relative higher expression of Foxo1 in CLP and early NK precursors may also contribute to the later NK cell proliferation and responsiveness, which warranties another separate study in the future.

Epidemiological analysis of 9,779 burn patients in China: An eight-year retrospective study at a major burn center in southwest China.

Burns are tissue injuries caused by high temperature, chemicals or electricity. Severe burns may involve all of the organs and tissues of the human body, leading to a series of pathophysiological processes and even death. The present study reviewed the clinical data of burn patients, including cases of burn-associated death, to provide evidence for the strategy of burn prevention. Basic information from 13,205 inpatients treated between January 1, 2009 and December 31, 2016 was extracted from the database of the Institute of Burn Research at Southwest Hospital (Chongqing, China). After excluding 3,426 inpatients who were not primarily treated for burns, 9,779 patients remained; among them, 68 cases (0.7%) had died as a direct consequence of the burns. Based on baseline data, the mortality rate, total body surface area of the burn (TBSA), age, sex, cause of injury and complications were analysed. In general, males accounted for a higher percentage than female burn patients. Of the patients, 95.54% had a TBSA of <50%, and the rate of mortality of the patients was increased when the TBSA was ≥50%. The major causes of injury were scalds (41.60%), fire (26.92%) and electricity (15.29%), and the majority of victims were 14 years or younger. With improvements in burn treatment technology in recent years, burn patient mortality was significantly reduced. Complications, including multiple organ failure and severe systemic infection, may reduce the survival rate of patients. The major risk factors for death included burns resulting from explosions, as well as shock, age (aged 0-1 or ≥50 years), greater TBSA and full-thickness burn area. With increasing length of stay at the hospital, patient mortality decreased. The renewal of treatment concepts and refined patient management contributed to the shorter LOS and lower mortality in 2015 and 2016.

Reconstruction and Functional Annotation of P311 Protein-Protein Interaction Network Reveals Its New Functions.

P311 is a highly conserved multifunctional protein. However, it does not belong to any established family of proteins, and its biological function has not been entirely determined. This study aims to reveal the unknown molecular and cellular function of P311. OCG (Overlapping Cluster Generator) is a clustering method used to partition a protein-protein network into overlapping clusters. Multifunctional proteins are at the intersection of relevant clusters. DAVID is an analytic tool used to extract biological meaning from a large protein list. Here we presented OD2 (OCG + DAVID + 2 human PPI datasets), a novel strategy to increase the likelihood to identify biological functions most pertinent to the multifunctional proteins. The principle of OD2 is that OCG prepares the protein lists from multifunctional protein relevant overlapping clusters, for a functional enrichment analysis by DAVID, and the similar functional enrichments, which occurs simultaneously when analyzing two human PPI datasets, are supposed to be the predicted functions. By applying OD2 to two reconstructed human PPI datasets, we supposed the function of the P311 in inflammatory responses, cell proliferation and coagulation, which were confirmed by the following biological experiments. Collectively, our study preliminarily found that P311 could play a role in inflammatory responses, cell proliferation and coagulation. Further studies are required to validate and elucidate the underlying mechanism.

Synthesis of graphene oxide-quaternary ammonium nanocomposite with synergistic antibacterial activity to promote infected wound healing.

BACKGROUND: Bacterial infection is one of the most common complications in burn, trauma, and chronic refractory wounds and is an impediment to healing. The frequent occurrence of antimicrobial-resistant bacteria due to irrational application of antibiotics increases treatment cost and mortality. Graphene oxide (GO) has been generally reported to possess high antimicrobial activity against a wide range of bacteria in vitro. In this study, a graphene oxide-quaternary ammonium salt (GO-QAS) nanocomposite was synthesized and thoroughly investigated for synergistic antibacterial activity, underlying antibacterial mechanisms and biocompatibility in vitro and in vivo. METHODS: The GO-QAS nanocomposite was synthesized through amidation reactions of carboxylic group end-capped QAS polymers with primary amine-decorated GO to achieve high QAS loading ratios on nanosheets. Next, we investigated the antibacterial activity and biocompatibility of GO-QAS in vitro and in vivo. RESULTS: GO-QAS exhibited synergistic antibacterial activity against bacteria through not only mechanical membrane perturbation, including wrapping, bacterial membrane insertion, and bacterial membrane perforation, but also oxidative stress induction. In addition, it was found that GO-QAS could eradicate multidrug-resistant bacteria more effectively than conventional antibiotics. The in vitro and in vivo toxicity tests indicated that GO-QAS did not exhibit obvious toxicity towards mammalian cells or organs at low concentrations. Notably, GO-QAS topically applied on infected wounds maintained highly efficient antibacterial activity and promoted infected wound healing in vivo. CONCLUSIONS: The GO-QAS nanocomposite exhibits excellent synergistic antibacterial activity and good biocompatibility both in vitro and in vivo. The antibacterial mechanisms involve both mechanical membrane perturbation and oxidative stress induction. In addition, GO-QAS accelerated the healing process of infected wounds by promoting re-epithelialization and granulation tissue formation. Overall, the results indicated that the GO-QAS nanocomposite could be applied as a promising antimicrobial agent for infected wound management and antibacterial wound dressing synthesis.

Nitric oxide induces epidermal stem cell de-adhesion by targeting integrin ß1 and Talin via the cGMP signalling pathway.

OBJECTIVE: Nitric oxide (NO) has emerged as a critical molecule in wound healing, but the mechanism underlying its activity is not well defined. Here, we explored the effect of NO on the de-adhesion of epidermal stem cells (ESCs) and the mechanism involved in this process. METHODS: The effects of NO on isolated human and mouse ESCs cultured in the presence of different concentrations of the NO donor S-nitroso-N-acetyl penicillamine (SNAP) were evaluated in cell de-adhesion assays mediated by integrin ß and collagen IV. Subsequently, changes in the expression of integrin ß1 and the phosphorylation of Talin in response to different doses of SNAP were detected by Western blot analysis and real-time PCR in vitro. Furthermore, the roles of various soluble guanylyl cyclase (sGC)- and protein kinase G (PKG)-specific inhibitors and agonists in the effects of NO on ESC de-adhesion, integrin ß1 expression and Talin phosphorylation were analysed. Moreover, the effects of NO on integrin ß1 expression and sGC/cGMP/PKG signalling-mediated wound healing were detected in vivo using 5-bromo-2-deoxyuridine (BrdU) label-retaining cells (LRCs) in a scald model and an excision wound healing model, respectively. RESULTS: SNAP promoted primary human and mouse ESC de-adhesion in a concentration-dependent manner in the integrin ß1-and collagen IV-mediated adhesion assay, and this effect was suppressed by the sGC and PKG inhibitors. Additionally, integrin ß1 expression and Talin phosphorylation at serine 425 (S425) were negatively correlated with SNAP levels, and this effect was blocked by the sGC and PKG inhibitors. Moreover, the roles of NO in integrin ß1 expression and cGMP signalling pathway-mediated wound healing were confirmed in vivo. CONCLUSION: Our data indicate that the stimulatory effects of NO on ESC de-adhesion related to integrin ß1 expression and Talin phosphorylation were mediated by the cGMP signalling pathway, which is likely involved in wound healing.

Polydimethylsiloxane incorporated with reduced graphene oxide (rGO) sheets for wound dressing application: Preparation and characterization.

Toward fabricating a novel multifunctional wound dressing material, we incorporated a series of contents of reduced graphene oxide (rGO) sheets into polydimethylsiloxane (PDMS) matrix to prepare the rGO-PDMS composite membrane and be used for wound dressing. The pore structure, dispersion of rGO, physical properties, water vapor transmission rate (WVTR), cytotoxicity and antibacterial activity were studied. Finally, the effect of the rGO-PDMS composite membrane on wound healing was investigated on a murine full-thickness skin wound model. The rGO-PDMS composite membrane exhibited bionic performance (ordered pore structure and suitable WVTR), improved mechanical properties, good compatibility and effective antibacterial activity. In vivo experiment indicated that the rGO-PDMS composite membrane could accelerate wound healing via enhancement of the re-epithelialization and granulation tissue formation. These findings suggest that rGO doping PDMS uniquely resulted in a multifunctional material for potential use in wound dressing.

Nitric oxide promotes epidermal stem cell proliferation via FOXG1-c-Myc signalling.

OBJECTIVE: Epidermal stem cells (ESCs) play a critical role in wound repair, but the mechanism underlying ESC proliferation is unclear. Here, we explored the effects of nitric oxide (NO) on ESC proliferation and the possible underlying mechanism. METHODS: The effect of NO (two NO donors, SNAP and spermine NONOate, were used) on cell proliferation was detected using cell proliferation and DNA synthesis assays. Thereafter, expression of FOXG1 and c-Myc induced by NO was determined by immunoblot analysis. pAdEasy-FOXG1 adenovirus and c-Myc siRNA plasmids were infected or transfected, respectively, into human ESCs to detect the effect of FOXG1 and c-Myc on NO-induced cell proliferation. Additionally, NO-induced ESC proliferation in vivo was detected by BrdU incorporation and a superficial second-degree mouse burn model. Moreover, the relationships among NO, FOXG1 and c-Myc were detected by western blotting, real-time PCR and dual luciferase assay. RESULTS: NO exerted a biphasic effect on ESC proliferation, and 100 µM SNAP and 10 µM spermine NONOate were the optimal concentrations to promote cell proliferation. Additionally, NO-promoted human ESC proliferation was mediated by FOXG1 and c-Myc in vitro and vivo. Furthermore, NO regulated FOXG1 expression through cGMP signalling, and NO-induced transcription of c-Myc was regulated by FOXG1-mediated c-Myc promoter activity. CONCLUSION: This study showed that the biphasic effect of NO on ESC proliferation as well as NO induced ESC proliferation were regulated by the cGMP/FOXG1/c-Myc signalling pathway, suggesting that NO may serve as a new disparate target for wound healing.

P311 Deficiency Leads to Attenuated Angiogenesis in Cutaneous Wound Healing.

P311 was identified to markedly promote cutaneous wound healing by our group. Angiogenesis plays a key role in wound healing. In this study, we sought to define the role of P311 in skin wound angiogenesis. It was noted that P311 was expressed in endothelial cells in the dermis of murine and human skin wounds. The expression of P311 was confirmed in cultured murine dermal microvascular endothelial cells (mDMECs). Moreover, it was found that knockout of P311 could attenuate the formation of tubes and motility of mDMECs significantly in vitro. In the subcutaneous Matrigel implant model, the angiogenesis was reduced significantly in P311 knockout mice. In addition, wound healing was delayed in P311 knockout mice compared with that in the wild type. Granulation tissue formation during the defective wound healing showed thinner and blood vessel numbers in wound areas in P311 knockout mice were decreased significantly. A reduction in VEGF and TGFß1 was also found in P311 KO mice wounds, which implied that P311 may modulate the exprssion of VEGF and TGFß1 in wound healing. Together, our findings suggest that P311 plays an important role in angiogenesis in wound healing.