摘要
目的探讨新西兰大白兔骨髓间充质干细胞(BMSC)同种异体移植促进预构皮瓣的血管再生,从而提高预构皮瓣存活率的可能性。方法体外分离、培养、鉴定并标记新西兰大白兔的BMSC。在实验兔腹部两侧构建预构皮瓣,两侧皮瓣随机分为实验组和对照组,实验组在股血管周围经皮注射已标记的BMSC悬液,对照组注射等量生理盐水。注射后第7天追踪观察BMSC移植后的成活情况,术后第14天掀起以股动静脉为轴心血管的岛状皮瓣,分别对两组的岛状皮瓣进行BrdU/vWF的免疫荧光双标检测;对岛状皮瓣中的VEGF进行Western Blot半定量分析;岛状皮瓣形成后第7天,观察两组皮瓣的成活情况。结果 BMSC异体移植后成活良好;实验组皮瓣内VEGF蛋白表达量明显高于对照组;实验组皮瓣内BrdU标记阳性细胞胞浆内有vWF信号;将预构皮瓣制成岛状皮瓣后第7天,实验组和对照组皮瓣存活率分别为(93.1±2.6)%、(51.5±7.5)%,P<0.05。结论异体移植BMSC可促进预构皮瓣的血管再生,提高预构皮瓣存活率。
Objective To evaluate the effects of bone marrow-derived mesenchymal stem cells（BMSC） transplantation on survival of prefabricated flap with increased neovascularization in a rabbit model.Methods BMSCs of New Zealand rabbits were isolated and cultured in vitro.The third passage cells were labeled before transplantation.Two prefabricated flaps of each rabbit were randomly allocated to experiment group or control group.In all animals,the subcutaneous tissue around the tunnel was injected with BMSCs in experiment group,or saline in equal amounts in control group.One week later,one flap from each group was selected to observe the viability of BMSCs.Two weeks later,abdominal island flap based solely on the implanted vessel was elevated.Three flaps were selected from each group to observe the expression of VEGF by Western Blot.Four flaps were selected from each group to observe the signal of vWF and BrdU by immuno-fluorescence histochemical staining.Results BMSCs were found viable in the flap from the experiment group.There was a significant increase in mean survival rate of prefabricated flaps from the experiment group compared to the control group： BMSCs（93.1±2.6）% vs.saline（51.5±7.5）%（P〈0.01）.Immuno-fluorescence histochemical staining found the signal of vWF in the cytoplasm of BrdU positive cells.Western Blot semi-quantitative analysis showed that the The VEGF protein level was found significantly higher in the tissues from the experiment group than that from the control group.Conclusion The findings from this study demonstrated that BMSCs transplantation could promote neovascularization and improve prefabricated flap survival.
出处
《组织工程与重建外科杂志》
2011年第1期11-15,共5页
Journal of Tissue Engineering and Reconstructive Surgery
基金
上海交通大学医学院附属第六人民医院院级基金(院-1349)
关键词
间充质干细胞
移植
预构皮瓣
血管内皮生长因子
Mesenchymal stem cells
Transplantation
Prefabricated flap
Vascular endothelial growth factor
