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目的构建经血管内皮生长因子165(VEGF165)转染的人成纤维细胞-脱细胞异种真皮替代物,并对其在促进深度烧伤患者创面愈合过程中的作用进行评价。方法将30例深Ⅱ度或Ⅲ度烧伤患者随机分为2组,每组15例。移植转染有VEGF165成纤维细胞-脱细胞异种真皮替代物组为实验组,单纯脱细胞异种真皮替代物移植组为对照组。采用分子生物学方法将扩增的VEGF165基因克隆入真核表达载体pcDNA3.1(+),在脂质体介导下将重组质粒转染至经传代培养的人成纤维细胞,再将转染后的人成纤维细胞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深度烧伤患者切痂后创面,对创面愈合情况进行观察并与单纯脱细胞真皮移植组进行比较。结果构建的真核表达载体pcDNA3.1(+)-VEGF165可成功转染人成纤维细胞,经转染的人成纤维细胞可顺利接种于脱细胞真皮形成成纤维细胞-脱细胞异种真皮替代物,替代物移植后实验组患者皮片成活率(90.25±5.39)%,对照组患者皮片成活率(83.98±3.63)%,两者有显著性差异(t=3.737,P<0.01)。实验组新生的毛细血管数量要明显多于对照组。结论经VEGF165转染的人成纤维细胞-脱细胞异种真皮替代物具有显著的促进深度烧伤患者创面愈合的作用。
Objective To construct a human dermal fibroblast - acellular dermal substitute transfected with vascular endothelial growth factor 165 (VEGF165) and evaluate its role in promoting wound healing in patients with deep burn. Methods Thirty patients with deep second degree or third degree burn were randomly divided into two groups (n = 15 in each group). Transplantation of VEGF165 fibroblast - acellular dermal dermal substitute group for the experimental group, simple acellular dermal graft for the control group. The amplified VEGF165 gene was cloned into the eukaryotic expression vector pcDNA3.1 (+) by molecular biology method. The recombinant plasmid was transfected into the subcultured human fibroblasts by liposome, and then transfected After the human fibroblasts were inoculated on the surface of porcine acellular dermis to obtain fibroblast-acellular dermal substitute, the substitute and autologous thin skin graft were transplanted to the wound surface after escharectomy, and the wound healing was observed And compared with pure acellular dermal transplantation group. Results The constructed eukaryotic expression vector pcDNA3.1 (+) - VEGF165 could be successfully transfected into human fibroblasts. The transfected human fibroblasts could be successfully inoculated into acellular dermal fibroblasts - acellular dermal dermal substitute (90.25 ± 5.39)% in the experimental group and 83.98 ± 3.63% in the control group (t = 3.737, P <0.01). The experimental group of newborn capillaries to be significantly more than the control group. Conclusion VEGF165 transfected human fibroblasts - acellular dermal dermal substitute has a significant role in promoting wound healing in patients with deep burn.