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目的:构建经血管内皮生长因子(VEGF)165转染的成纤维细胞-脱细胞异种真皮替代物,观察其在促进大鼠深度创面愈合过程中的作用。方法:采用分子生物学方法将扩增的VEGF165基因克隆人真核表达载体pcDNA3.1(+),在脂质体介导下将重组质粒转染至经传代培养的鼠成纤维细胞(NIH/3T3),再将转染后的NIH/3T3接种于猪脱细胞真皮表面得到成纤维细胞-脱细胞异种真皮替代物;而后将此替代物与自体薄皮片移植于SD大鼠深度创面(实验组),对动物创面愈合情况进行观察。单纯脱细胞异种真皮替代物移植大鼠作为对照组。结果:构建的真核表达载体pcDNA3.1(+)-VEGF165可成功转染至鼠成纤维细胞,移植后实验组皮片成活率(93.76±3.40)%,对照组皮片成活率(85.01±1.74)%,两者差异有显著性(t=8.873,P<0.01)。组织学观察显示,移植第4周实验组毛细血管数量明显多于对照组。结论:成功构建了经VEGF165转染的成纤维细胞-脱细胞异种真皮替代物,且其具有明显的促进动物创面愈合作用。
OBJECTIVE: To construct a fibroblast-derived acellular dermal substitute transfected with vascular endothelial growth factor (VEGF) 165 and observe its role in promoting deep wound healing in rats. METHODS: The amplified VEGF165 gene was cloned into human eukaryotic expression vector pcDNA3.1 (+) by molecular biology method. The recombinant plasmid was transfected into NIH / 3T3). The transfected NIH / 3T3 was inoculated on the surface of porcine acellular dermis to obtain fibroblast-decellularized xenograft dermis substitute. Then, the substitute and autologous thin skin graft were transplanted into SD rat deep wound (experimental group ), Observed the wound healing of animals. Simple acellular dermal substitutes were transplanted in rats as a control group. Results: The eukaryotic expression vector pcDNA3.1 (+) - VEGF165 could be successfully transfected into murine fibroblasts. The graft survival rate of the experimental group was 93.76 ± 3.40% and that of the control group was 85.01 ± 1.74)%, the difference was significant (t = 8.873, P <0.01). Histological observation showed that in the fourth week of transplantation, the number of capillaries in the experimental group was significantly more than that in the control group. CONCLUSION: The fibroblast-acellular dermal dermal substitute transfected with VEGF165 has been successfully constructed and has the obvious effect of promoting wound healing in animals.