Gaoxing Luo

Investigating the role of P311 in the hypertrophic scar.

The mechanisms of hypertrophic scar formation are not fully understood. We previously screened the differentially expressed genes of human hypertrophic scar tissue and identified P311 gene as upregulated. As the activities of P311 in human fibroblast function are unknown, we examined the distribution of it and the effects of forced expression or silencing of expression of P311. P311 expression was detected in fibroblast-like cells from the hypertrophic scar of burn injury patients but not in peripheral blood mononuclear cells, bone marrow mesenchymal stem cells, epidermal cells or normal skin dermal cells. Transfection of fibroblasts with P311 gene stimulated the expression of alpha-smooth muscle actin (α-SMA), TGF-β1 and α1(I) collagen (COL1A1), and enhanced the contraction of fibroblast populated collagen lattices (FPCL). In contrast, interference of fibroblast P311 gene expression decreased the TGF-β1 mRNA expression and reduced the contraction of fibroblasts in FPCL. These results suggest that P311 may be involved in the pathogenesis of hypertrophic scar via induction of a myofibroblastic phenotype and of functions such as TGF-β1 expression. P311 could be a novel target for the control of hypertrophic scar development.

Analyses of differential proteome of human synovial fibroblasts obtained from arthritis

There is mounting evidence indicating that the synovial fibroblasts (SFs) contribute to the pathogenesis of rheumatoid arthritis (RA). The present study showed the differential proteins expression pattern of SFs from patients with RA or osteoarthritis (OA) and healthy control. Cellular proteins of cultured SFs were subjected to 2-DE and visualized by silver nitrate staining. A total of 49 spots that were statistically and differentially overexpressed in RA or OA in comparison to healthy ones were identified by MALDI-TOF-MS, and 25 proteins were successfully identified. Western blot was used to further verify some of the differential proteins. These proteins included enzymatic and structural proteins, signal transduction proteins, calcium binding protein, etc. From all of the identified proteins, a number of proteins have been implicated that involved in the healthy or pathological SFs function (e.g., S100A4, S100A10, cathepsin D) or that have potential diagnostic and prognostic value for RA (α-enolase and TPI) or that may be the new therapeutic targets (Annexin, SOD, PRX).

Controlled water vapor transmission rate promotes wound-healing via wound re-epithelialization and contraction enhancement

A desirable microenvironment is essential for wound healing, in which an ideal moisture content is one of the most important factors. The fundamental function and requirement for wound dressings is to keep the wound at an optimal moisture. Here, we prepared serial polyurethane (PU) membrane dressings with graded water vapor transmission rates (WVTRs), and the optimal WVTR of the dressing for wound healing was identified by both in vitro and in vivo studies. It was found that the dressing with a WVTR of 2028.3 ± 237.8 g/m2·24 h was able to maintain an optimal moisture content for the proliferation and regular function of epidermal cells and fibroblasts in a three-dimensional culture model. Moreover, the dressing with this optimal WTVR was found to be able to promote wound healing in a mouse skin wound model. Our finds may be helpful in the design of wound dressing for wound regeneration in the future.

A self-healing, re-moldable and biocompatible crosslinked polysiloxane elastomer

The thermally healable polysiloxane elastomers were successfully prepared by cross-linking polydimethylsiloxane bearing maleimide pendants with furan-end functionalized siloxane via the Diels-Alder (DA) reaction. The elastomers with good mechanical properties show excellent self-healing and remoldability functions due to the thermally reversible feature of the DA reaction. The molecular mechanism of self-healing was confirmed by in situ structural characterization. Moreover, the biocompatibility of the polysiloxane elastomer containing DA bonds is found to be fairly good by cytotoxicity evaluation and animal subcutaneous experiments, suggesting its potential applications in the biomedical field as artificial skin and scaffolds for tissue engineering.

Preparation of Microporous Silicone Rubber Membrane with Tunable Pore Size via Solvent Evaporation-Induced Phase Separation

Silicone rubber membrane with ordered micropores in the surface was prepared by means of the solvent evaporation-induced phase separation. A ternary solution including liquid silicone rubber precursor, liquid paraffin, and hexane was cast to form a film with a two-phase structure after the hexane was evaporated. The micropores were generated by removing liquid paraffin phase in the cured silicone rubber film. The effects of the liquid paraffin concentration, casting temperature, initial casting solution thickness, air circulation, and addition of surfactant Span-80 on the pore structure in the membrane surface were investigated. The average pore size increases with increasing liquid paraffin concentration or the initial casting solution thickness. The formation of pore structure in the membrane surface is related to the phase separation and thus the phase separation process of the casting solution surface was in situ observed using the digital microscope. The formation mechanism of pore is attributed to a nucleation, growth, and coalescence process of liquid paraffin phase in the membrane surface.

Self-healing poly(siloxane-urethane) elastomers with remoldability, shape memory and biocompatibility

Thermally healable poly(siloxane-urethane) elastomers (PDMS-DA-PU and PDMS/PCL-DA-PU) with Diels–Alder (DA) bonds alternately located in the polymer chains between crosslinking points were prepared based on self-made soft segments, isocyanate terminated polydimethylsiloxane (NCO-PDMS-NCO) or a mixture of NCO-PDMS-NCO and polycaprolactone diol. The elastomers show good self-healing and remoldable behaviors via DA and retro-DA reactions, which was confirmed by UV-vis and 1H NMR. In order to enhance the mechanical properties, the PCL segment was introduced into the chain of poly(siloxane-urethane). The microphase separation structure and covalent crosslinked network endow the polymers with good tensile strength, elasticity and shape recovery ability. Moreover, the biocompatibility of the poly(siloxane-urethane) elastomer was found to be fairly good by cytotoxicity evaluation and animal wound healing experiments, suggesting its potential application in biomedical fields such as artificial skin with a self-healing function.

Nitric Oxide Enhances Keratinocyte Cell Migration by Regulating Rho GTPase via cGMP-PKG Signalling

Objective Nitric oxide (NO) has been shown to improve wound healing, but the mechanism underlying this function is not well defined. Here, we explored the effect of NO on the migration of a human keratinocyte cell line (HaCaT) and its possible mechanism. Methods The effects of NO on HaCaT cells in the presence of different concentrations of the NO donor sodium nitroprusside (SNP) were evaluated in a cell migration assay. Subsequently, the cytoskeleton reorganization of cultured HaCaT cells stained with rhodamine-phalloidin was observed with a confocal laser scanning microscope. The mRNA expression and active proteins of CDC42, Rac1 and RhoA in the cultured cells were determined via RT-PCR and pull-down assays, respectively. Furthermore, the roles of various inhibitors or agonists specific to cGMP, PKG and CDC42, Rac1, RhoA in the effects of NO on HaCaT cell migration, F-actin stress fibre formation, and Rho GTPase expression were observed. Results It was also found HaCaT cell migration was increased by SNP in a dose-dependent manner, and the other two NO donors either spermine NONOate or SNAP had almost the same effects on HaCat cell migrations. The formation of F-actin stress fibres in SNP-treated HaCaT cells was increased. The mRNA expression and the active proteins of CDC42, Rac1 and RhoA were found to be upregulated after SNP treatment. Similar effects were observed after the cells were treated with a cGMP or PKG agonist. Additionally, the SNP-mediated upregulation of the mRNA expression and the active proteins of CDC42, Rac1 and RhoA were inhibited by the addition of an inhibitor of cGMP or PKG. Moreover, the SNP-mediated promoting effects of migration and cytoskeleton reorganization were inhibited by treatment with inhibitors of cGMP, PKG, CDC42, Rac1 and RhoA respectively. Conclusion Our data indicated that the stimulatory effects of NO on cell migration of HaCaT cells are mediated by the cGMP signalling pathway via the upregulation of Rho-GTPase expression, which might promote cytoskeleton reorganization.

Platelet-Derived Growth Factor Receptor Beta: A Novel Urinary Biomarker for Recurrence of Non-Muscle-Invasive Bladder Cancer

Non-muscle-invasive bladder cancer (NMIBC) is one of the most common malignant tumors in the urological system with a high risk of recurrence, and effective non-invasive biomarkers for NMIBC relapse are still needed. The human urinary proteome can reflect the status of the microenvironment of the urinary system and is an ideal source for clinical diagnosis of urinary system diseases. Our previous work used proteomics to identify 1643 high-confidence urinary proteins in the urine from a healthy population. Here, we used bioinformatics to construct a cancer-associated protein-protein interaction (PPI) network comprising 16 high-abundance urinary proteins based on the urinary proteome database. As a result, platelet-derived growth factor receptor beta (PDGFRB) was selected for further validation as a candidate biomarker for NMIBC diagnosis and prognosis. Although the levels of urinary PDGFRB showed no significant difference between patients pre- and post-surgery (n = 185, P>0.05), over 3 years of follow-up, urinary PDGFRB was shown to be significantly higher in relapsed patients (n = 68) than in relapse-free patients (n = 117, P<0.001). The levels of urinary PDGFRB were significantly correlated with the risk of 3-year recurrence of NMIBC, and these levels improved the accuracy of a NMIBC recurrence risk prediction model that included age, tumor size, and tumor number (area under the curve, 0.862; 95% CI, 0.809 to 0.914) compared to PDGFR alone. Therefore, we surmise that urinary PDGFRB could serve as a non-invasive biomarker for predicting NMIBC recurrence.

Inhalation injury in southwest China—The evolution of care

AIM This study aims to review the changes in management of inhalation injury and the associated reduction in mortality over the past 2 decades. METHODS The records of burn patients with inhalation injury hospitalised in our institute from 1986 to 2005 were retrospectively analysed. The incidence of inhalation injury and the associated mortality were analysed. Meanwhile, the relationship of inhalation injury with age, total burn area, tracheostomy intubation and mechanical ventilation were studied. RESULTS The incidence of inhalation injury was 8.01% in the total 10 608 hospitalised burn patients during the 20 years surveyed. Inhalation injury was always associated with large-sized burn and was more common in adults. The incidence of tracheostomy and mechanical ventilation increased from 39.46 and 30.28% in the period from 1986 to 1995 to 70.12 and 39.74% from 1996 to 2005, respectively. The overall mortality of inhalation-injured burn patients was 15.88% compared with 0.82% of the non-inhalation group. The mortality of the burn patients with inhalation injury dropped from 25.29% during the first 10 years to 11.71% during the second decade (p<0.01). Mortality secondary to inhalation injury as the lead cause decreased from 14.56 to 6.29% (p<0.01). CONCLUSION The care of inhalation injury has made significant progress over the past 2 decades. The early diagnosis of inhalation injury, early airway control and pulmonary function assistance with mechanical ventilation contribute to the reduction of mortality.

Nano-silver-decorated microfibrous eggshell membrane: processing, cytotoxicity assessment and optimization, antibacterial activity and wound healing

An ideal wound dressing can both promote wound healing and prevent bacterial infection. Here, we report a potential dressing prepared by incorporating an optimized concentration of silver nanoparticles (AgNPs) into the microfibers of a natural eggshell membrane (EM) using environmentally friendly and mussel-inspired dopamine. Briefly, acid-treated EM was used as a porous membrane for polydopamine-reduced AgNPs synthesis. To obtain the optimal cytocompatible silver concentration, cellular attachment and MTT assay were performed with different concentrations of AgNPs. The morphology of the EM and AgNPs was confirmed by scanning electronic microscopy, scanning transmission electronic microscopy and Fourier transform infrared spectroscopy. The synthesized EM/AgNPs exhibited steady and safe AgNPs release, which was further tested for antibacterial activity against Escherichia coli and Staphylococcus aureus by disc diffusion method and bacterial suspension assay. Finally, in a murine full-thickness skin wound model, we found that EM/AgNPs could promote re-epithelialization, granulation tissue formation and wound healing via enhancing cell proliferation, as demonstrated by the expression of proliferating cell nuclear antigen (PCNA), and controlling inflammation response, as demonstrated by the expression of interleukin-1β (IL-1β). These findings suggest that EM/AgNPs may have a promising application in wound management.