转基因骨髓间充质干细胞修复严重韧带损伤的实验观察  

Experimental study of repairing severe ligament injury with transgenetic bone mesenchymal stem cells

作　　者：周盛源[1,2] 朱巍[1] 陈雄生[1] 许政[1] 钱闯[1] 刘晓东[1] 

机构地区：[1]上海长征医院骨科,250021 [2]上海市闸北区中心医院骨科

出　　处：《中华医学杂志》2014年第19期1494-1498,共5页National Medical Journal of China

基　　金：国家自然科学基金（81201378）;国家“863”高技术研究发展计划项目（2008AA022418）;上海市科委自然科学基金（12ZR1439000）;上海市卫生局青年科研基金（2011Y127）

摘　　要：目的 评价转染碱性成纤维细胞生长因子（bFGF）基因的骨髓间充质干细胞（BMSCs）对严重韧带损伤的修复作用.方法 通过腺病毒将bFGF基因转入BMSCs,并标记增强型绿色荧光蛋白（eGFP）.建立新西兰大白兔髌韧带损伤模型,在断裂韧带缝合后,根据局部细胞移植情况将手术组分为转基因BMSCs组（组1）、BMSCs组（组2）、无细胞组（组3）、正常韧带组（组4）.术后不同时间点观察韧带损伤修复区转基因BMSCs的分布,bFGF表达及韧带特异性细胞外基质：Ⅰ型胶原、Ⅲ型胶原、fibronectin、α-SMA和vimentin的表达;组织学观察各组韧带损伤愈合情况;生物力学检测评价各组的生物力学强度.结果 转基因BMSCs组术后1个月内在韧带修复区内可持续观察到转基因BMSCs的存在,组1术后3～14 d bFGF表达明显高于对照组（P ＜0.001）,其中移植后7d为表达高峰.术后1个月内转基因BMSCs组各韧带特异性细胞外基质基因表达明显强于对照组（P＜0.001）,免疫组化呈强阳性表现.术后3个月HE染色见转基因BMSCs组韧带损伤修复区胶原纤维排列及致密度均优于对照组,生物力学检测表明转基因BMSCs组拉力载荷明显强于对照组[组1～4分别为（13.5±1.3）、（8.6±2.0）、（8.4±2.7）、（20.5±4.9）N,P＜0.05].结论 转染bFGF基因的BMSCs能明显促进韧带损伤愈合,增加韧带愈合强度.Objective To evaluate the efficacies of transgenetic bone mesenchymal stem cells （BMSCs） carring basic fibroblast growth factor （bFGF） gene on repairing severe ligament injury.Methods The genes of bFGF and enhanced green fluorescent protein （eGFP） were transfected into BMSCs through adenovirus vector.According to different cell transplantations,the operated rabbits were divided into 3groups of transgenetic BMSCs,BMSCs and no cells.And group 4 was normal ligament group.The distribution of transgenetic BMSCs in ligament injury area,the expression of bFGF and ligament specific extracellular matrix,i.e.type Ⅰ collagen,type Ⅲ collagen,fibronectin,alpha-smooth muscle actin and vimentin,histology and biomechanics of ligament healing were recorded at different timepoints.Results The transgenetic BMSCs could be observed in injury area within one month.The expression of bFGF in the transgenetic BMSCs group was significantly higher than that in the control groups at Days 3-14 （P 〈0.001） and peaked at Day 7.In the transgenetic BMSCs group,the gene expression of ligament specific extracellular matrix was significantly higher than that in the control groups within one month （P 〈 0.001）.And immunohistochemistry showed strong positive expression.After 3 months,in group 1,hematoxylin and eosin staining showed the arrangement and density of collagen fibers of healing area were better than that in the control groups.And biomechanical testing showed that tensile load were stronger than that in the control groups （P 〈 0.05）.Conclusion Transgenetic BMSCs carrying bFGF gene can obviously promote ligament injury healing and increase ligament healing strength.

关 键 词：韧带 创伤和损伤 成纤维细胞生长因子 碱性 干细胞 

分 类 号：R686.5[医药卫生—骨科学]