转染TGF-β1的骨髓间充质干细胞移植对预构皮瓣存活影响的实验研究  

Effect of transplantation of bone marrow mesenchymal stem cells transfected with TGF-β1 on survival of prefabricated flap: an experimental study

作　　者：尹朝奇[1] 周建大[1] 罗成群[1] 贺全勇[1] 陶如意[1] 苏红辉[1] 张红莲[1] 陈佳[1] 李萍[1] 王少华[1] 

机构地区：[1]中南大学湘雅三医院烧伤整形科,湖南长沙410013

出　　处：《中国普通外科杂志》2016年第6期875-881,共7页China Journal of General Surgery

基　　金：湖南省科学技术厅科技计划资助项目(2011SK3243)

摘　　要：目的:探讨TGF-β1转染骨髓间充质干细胞(MSC)移植对预构皮瓣存活的影响及机制。方法:分离并培养大鼠MSC,采用脂质体介导技术将真核表达载体pc DNA3.1(+)/TGF-β1转染至MSC,对转染后MSC的表型及体外成管进行鉴定;选取12只SD大鼠,于其背部两侧对称建立5 cm×1 cm随意皮瓣模型,于两侧皮瓣下分别注射转染后的MSC(实验侧)与溶媒(对照侧),观察皮瓣存活情况计算皮瓣存活率,免疫组化染色及HE染色检测皮瓣血管密度。结果:流式细胞仪分析表明,转染TGF-β1后MSC仍具有MSC的特性,且具有良好的体外成管功能。注射转染TGF-β1的MSC的实验侧皮瓣存活率明显高于对照侧(82.83%vs.61.33%,P<0.05),同时皮瓣内毛细血管的密度明显也明显高于对照侧(19.69个vs.8.19个,P<0.05)。结论:转染TGF-β1的MSC可以促进皮瓣内毛细血管生成,提高皮瓣的存活率。Objective：The investigate the effect of transplantation of bone marrow mesenchymal stem cells （MSCs） transfected with TGF-β1 on survival of prefabricated lfap and the mechanism. Methods：The rat MSCs were isolated and cultured, and then transfected with pcDNA3.1 （＋）/TGF-β1 eukaryotic expression vectors through liposome delivery system. hTe phenotype and tube formation ability in vitro of the MSCs atfer transfection were determined. Twelve SD rats were used, and random lfaps with 5 cm in length and 1 cm in width were established on the symmetric parts of the back of the rats, and then the bottom of the lfap on both sides was injected with MSCs atfer transfection （experimental side） or vehicle （control side）, respectively. hTe survival rate of lfap was calculated and the vessel density in the lfap was determined by HE andimmunohistochemical staining. Results：Flow cytometry analysis showed that bone marrow stem cells transfected with TGF-β1 exhibited the characteristics of the na？ve MSCs, and had better tube formation ability in vitro. In the lfaps on experimental side injected with MSCs transfected with TGF-β1, the survival rate was signiifcantly higher than that in the lfaps on control sides （82.83%vs. 61.33%, P〈0.05）, and meanwhile, the capillary density was signiifcantly greater than that in the lfaps on control sides （19.69 vs. 8.19, P〈0.05）. Conclusion：Transplantation of MSCs transfected with TGF-β1 can promote angiogenesis in skin flaps and thereby improve the survival of the lfaps.

关 键 词：间质干细胞移植 转化生长因子β1 外科皮瓣 毛细血管 

分 类 号：R662.1[医药卫生—外科学]