骨形态发生蛋白4基因修饰的脂肪来源干细胞复合藻酸钙凝胶支架修复关节软骨的实验研究  

Repair of Articular Cartilage Defects with Bone Morphogenetic Protein 4 Gene-Enhanced Adipose-derived Stem Cells Combined with Calcium Alginate Gel Scaffold

作　　者：陈临新[1] 石媛媛[1] 张辛[1] 王健全[1] 敖英芳[1] 

机构地区：[1]北京大学第三医院运动医学研究所,北京100191

出　　处：《中国运动医学杂志》2016年第3期234-239,268,共7页Chinese Journal of Sports Medicine

基　　金：国家自然科学基金主任基金项目(81341053)

摘　　要：目的：观察骨形态发生蛋白4（BMP4）修饰的脂肪来源干细胞（ADSCs）复合藻酸钙凝胶支架修复猪膝关节软骨缺损的实验效果。方法：分离并培养猪ADSCs。构建表达BMP4的腺病毒载体系统,并成功转染ADSCs,通过免疫荧光显微镜及流式细胞学观察其转染效率。在体外,通过ELISA法检测BMP4基因修饰ADSCs后软骨相关蛋白的表达情况。选取中国实验用小型猪24只（雄性,6个月,25～30 kg）,随机分为4组：组1为空白对照组,即单纯软骨缺损组;组2为单纯藻酸钙凝胶支架修复组;组3为ADSCs复合藻酸钙凝胶修复组;组4为BMP4基因修饰的ADSCs复合藻酸钙支架修复软骨缺损组。分别于术后12周、24周取材,通过组织形态学方法来评估缺损修复情况。结果：体外实验证实,携带BMP4的腺病毒能够成功转染ADSCs,并诱导其表达软骨组织特异性蛋白Sox9和Col II。同时体内实验证实BMP4基因修饰的ADSCs复合藻酸钙凝胶支架可修复小型猪膝关节软骨缺损。结论：体内外实验证实,BMP4基因修饰的ADSCs复合藻酸钙凝胶支架可用于修复猪关节软骨缺损,为软骨损伤修复提供了一种选择。Objective To observe the efficacy of adipose-derived stem cells（ADSCs）transfected with bone morphogenetic protein 4（BMP4）combined with calcium alginate gel scaffold for repair of articular cartilage defects in a porcine model. Methods The ADSCs after isolation,cultivation,and transfection with BMP4 were expressed by a recombinant adenovirus vector system. The transfection efficiency was observed through immunofluorescence microscopy and flow cytometry. Cartilage-related proteins were detected by ELISA assay after BMP4 with ADSCs in vitro. Articular cartilage defects were established in 24 miniature pigs（male, 6-month old,25-30 kg）and then the pigs were divided into cartilage defect group,purely Ca Alg hydrogel repair group,Ca Alg hydrogel containing ADSCs repair group,and Ca Alg hydrogel containing BMP4-ADSCs repair group. The repair of the defect was evaluated by histology at 12 and 24 weeks after surgery. Results BMP4 carrying adenovirus can successfully transfect to ADSCs and the expression levels of genes such as BMP4,Sox9, and Col II increase in vitro. And the articular osteochondrogenic defects are well repaired by ADSCs transfected with BMP4 combined with calcium alginate gel scaffold in the porcine model. Conclusion The method introduced in this study provides an alternative selection for repair of cartilage defects.

关 键 词：骨形态发生蛋白4 脂肪来源干细胞 软骨缺损 

分 类 号：R687.3[医药卫生—骨科学] R318.08[医药卫生—外科学]