Regeneration of mature dermis by transplanted particulate acellular dermal matrix in a rat model of skin defect wound

Native mammalian extracellular matrix (ECM) has been made in various forms including particles, sheet and mesh which are appropriate for site-specific applications. The ECM particles are usually created by homogenization method and have a wider size distribution. This needs to be improved to produce more uniform ECM particles. In present study, we had successfully developed a method for preparing particulate acellular dermal matrix (PADM) in different gauges. The resultant PADM was approaching a rectangular parallelepiped or cubic shape, with a better or narrower size distribution than other ECM particles in previous reports. It also retained ultrastructure and functional molecules of native ECM. In vivo performances were evaluated after implantation of PADM in an acute full-thickness skin defect wound in rats. Histological analysis showed that allogeneic PADM used as dermal regeneration template could facilitate maturation and improving collagen bundle structure of regenerated dermis at the endpoint of 20 weeks post-surgery. The PADM could be used for further investigation in analyzing the impacts of cellularly and/or molecularly modified PADM on soft tissue regeneration

Inhibitory Effect of Curcumol on Jak2-STAT Signal Pathway Molecules of Fibroblast-Like Synoviocytes in Patients with Rheumatoid Arthritis

Hyperplasia of synovial membrane in rheumatoid arthritis (RA) is a critical pathological foundation for inducing articular injury. The janus kinase and signal transducer and activator of transcription (Jak-STAT) pathway plays a critical role in synovial membrane proliferation induced by platelet-derived growth factor (PDGF). To explore the anti-cell proliferation mechanism of curcumol, a pure monomer extracted from Chinese medical plant zedoary rhizome, the changes of Jak2-STAT1/3 signal pathway-related molecules in synoviocytes were observed in vitro. In this study, the fibroblast-like synoviocytes (FLS) in patients with RA were collected and cultured. The following parameters were measured: cell proliferation (WST-1 assay), cell cycles (fluorescence-activated cell sorting, FACS), STAT1 and STAT3 activities (electrophoretic mobility shift assay, EMSA), and the protein expressions of phosphorylated Jak2, STAT1, and STAT3 (Western blot). It was shown that curcumol could inhibit the RA-FLS proliferation and DNA synthesis induced by PDGF-BB in a dose-dependent manner in vitro. The transcription factors activities of STAT1 and STAT3 were obviously elevated after PDGF-BB stimulation (P < 0.05). Super-shift experiments identified the STAT1 or STAT3 proteins in the complex. Furthermore, the different concentration curcumol could downregulate the DNA binding activities of STAT1 and STAT3 (P < 0.05) and inhibit the phosphorylation of Jak2 while it had no effect on the protein expressions of STAT1 and STAT3. Positive correlations were found between changes of cell proliferation and DNA-binding activities of STAT1 and STAT3, respectively (P < 0.01). In conclusion, curcumol might suppress the FLS proliferation and DNA synthesis induced by PDGF-BB through attenuating Jak2 phosphorylation, downregulating STAT1 and STAT3 DNA-binding activities, which could provide theoretical foundation for clinical treatment of RA

Determination of nuclear transcription factor activity using a modified electrophoretic mobility shift assay

In this work, several major procedures of the electrophoretic mobility shift assay (EMSA) were modified including swift extraction of the nucleic protein, labeling of the probe and radioautography. The modified assay required shorter time, simplified the nucleic protein extraction, increased the radioactivity of the labeling probe, skipped the tedious process of gel drying, and produced clear images. Its results were comparable, reproducible and stable. It thus has merited for wide application

Determination of nuclear transcription factor activity using a modified electrophoretic mobility shift assay

In this work, several major procedures of the electrophoretic mobility shift assay (EMSA) were modified including swift extraction of the nucleic protein, labeling of the probe and radioautography. The modified assay required shorter time, simplified the nucleic protein extraction, increased the radioactivity of the labeling probe, skipped the tedious process of gel drying, and produced clear images. Its results were comparable, reproducible and stable. It thus has merited for wide application.A determinação da alteração na mobilidade eletroforética (EMSA), o método de mais ampla utilização para o estudo das interações proteínaácidos nucléicos, é tediosa e difícil de dominar. De acordo com os protocolos dacumentados e com base em nossa prática, nós modificamos os diversos processos principais dessa determinação incluindo no que diz respeito a extração de proteiínas nucleicas, marcação das provas e radioautografia. A determinação modificada requer menor tempo, simplifica a extração de ácidos nucleicos, eleva a radioatividade da prova marcada, evita o processo tedioso de secagem do gel e produz claras imagens. Seus resultados são comparáveis, reproduzíveis e estáveis, merecendo, desse modo, ampla aplicação

Effect of mixed transplantation of autologous and allogeneic microskin grafts on wound healing in a rat model of acute skin defect.

The treatment of extensive thermal injuries with insufficient autologous skin remains a great challenge to burn surgeons. In this study, we investigated the influence of the ratio of autologous and allogeneic tissue in mixed microskin grafts on wound healing in order to develop an effective method for using limited donor skin to cover a large open wound. Four different mixtures were tested: autologous microskin at an area expansion ratio of 10∶1 with allogeneic microskin at an area expansion ratio of 10∶1 or 10∶3 and autologous microskin at an expansion ratio of 20∶1 with allogeneic microskin at an expansion ratio of 20∶3 or 20∶6. Wound healing, wound contraction, and integrin β1 expression were measured. Mixed microskin grafting facilitated wound healing substantially. The mixture of autologous microskin at an expansion ratio of 10∶1 with the same amount of allogeneic microskin achieved the most satisfactory wound healing among the 4 tested mixtures. Histological examination revealed the presence of obviously thickened epidermis and ectopic integrin β1 expression. Keratinocytes expressing integrin β1 were scattered in the suprabasal layer. Higher levels of integrin β1 expression were associated with faster wound healing, implying that ectopic expression of integrin β1 in keratinocytes may play a pivotal role in wound healing. In conclusion, this study proves that this new skin grafting technique may improve wound healing

The progress of Chinese burn medicine from the Third Military Medical University—in memory of its pioneer, Professor Li Ao

Abstract Professor Li Ao was one of the founders of Chinese burn medicine and one of the most renowned doctors and researchers of burns in China. He established one of the Chinese earliest special departments for burns at Third Military Medical University (TMMU) in 1958. To memorialize Professor Li Ao on his 100th birthday in 2017 and introduce our extensive experience, it is our honor to briefly review the development and achievement of the Chinese burn medicine from TMMU. The epidemiology and outcomes of admitted burn patients since 1958 were reviewed. Furthermore, main achievements of basic and clinical research for the past roughly 60 years were presented. These achievements mainly included the Chinese Rule of Nine, fluid resuscitation protocol, experience in inhalation injury, wound treatment strategies, prevention and treatment of burn infections, nutrition therapy, organ support therapies, and rehabilitation. The progress shaped and enriched modern Chinese burn medicine and promoted the development of world burn medicine

Changes in the histological appearance and epidermal thickness of the healed wounds.

<p>Aa shows the normal skin in Sprague-Dawley (SD) rats; Ab, Ac, and Ad show the histological appearance of the healed wounds at 2 post-graft weeks (PGWs) in groups II,III, and IV, respectively, in Experiment A (hematoxylin and eosin; scale bar, 50 µm). B, C, and D show the changes from 2 to 4 PGWs in the epidermal thickness of the healed wounds in 3 groups in Experiment A. E and F show the changes from 3 to 4 PGWs in the epidermal thickness of the healed wounds in 2 groups from Experiment B (µm, ± s), * p<0.05.</p

Schematic diagram of measurement of the epidermal thickness and the gross appearance during wound healing.

<p>In <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0085672#pone-0085672-g002" target="_blank">Figure 2A, L, A, and H</a> represented the length, area, and average thickness, respectively, of the selected epidermal region. H was calculated according to the first mean value theorem for integration: H = A÷L. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0085672#pone-0085672-g002" target="_blank">Figure 2B</a> shows the gross appearance of wound healing in group III in Experiment A at 3 post-graft weeks (PGWs). The red arrows indicate desquamation. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0085672#pone-0085672-g002" target="_blank">Figure 2C</a> shows the gross appearance of wound healing in group I in Experiment A at 2 PGWs. The blue arrows show fresh granulation tissue revealed by the shedding of the allogeneic microskin, the black arrows show the remaining microskin, and the yellow arrows show the de novo epithelium at the wound edge.</p

Wound contraction rates after mixed microskin transplantation in Experiments A and B.

<p>A, B, and C show the wound contraction rates at 2, 3, and 4 post-graft weeks (PGWs), respectively, in Experiment A (* p<0.05).</p