文章摘要

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

作者: 1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 中南大学湘雅三医院 烧伤整形科，湖南 长沙 410013
通讯: 尹朝奇 Email: yinchaoqi@163.com
DOI: 10.3978/.10.3978/j.issn.1005-6947.2016.06.017
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pcDNA3.1（+）/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。
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

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

Authors: 1YIN Chaoqi, 1ZHOU Jianda, 1LUO Chengqun, 1HE Quanyong, 1TAO Ruyi, 1SU Honghui, 1ZHANG Honglian, 1CHEN Jia, 1LI Ping, 1WANG Shaohua
1 Department of Burn and Plastic Surgery, the Third Xiangya Hospital, Central South University, Changsha 410013, China

CorrespondingAuthor:YIN Chaoqi Email: yinchaoqi@163.com

Abstract

Objective: The investigate the effect of transplantation of bone marrow mesenchymal stem cells (MSCs) transfected with TGF-β1 on survival of prefabricated flap 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. The phenotype and tube formation ability in vitro of the MSCs after transfection were determined. Twelve SD rats were used, and random flaps 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 flap on both sides was injected with MSCs after transfection (experimental side) or vehicle (control side), respectively. The survival rate of flap was calculated and the vessel density in the flap was determined by HE and immunohistochemical staining. Results: Flow cytometry analysis showed that bone marrow stem cells transfected with TGF-β1 exhibited the characteristics of the naïve MSCs, and had better tube formation ability in vitro. In the flaps on experimental side injected with MSCs transfected with TGF-β1, the survival rate was significantly higher than that in the flaps on control sides (82.83% vs. 61.33%, P<0.05), and meanwhile, the capillary density was significantly greater than that in the flaps 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 flaps.
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