转化生长因子-β3与基质金属蛋白酶抑制剂-2联合转染兔骨髓间充质干细胞复合丝素蛋白/壳聚糖生物支架修复兔软骨缺损  

Resurfacing rabbit＇s cartilage defects using mesenchymal stem cells co-transfected with transforming growth factor-β3 and matrix metalloproteinases inhibitors-2 by adeno-associated virus embedded in Silk fibroin/chitosan scaffolds

作　　者：孟飞[1] 吕成昱[1] 张海宁[1] 申成凯[1] 冯尚祥 

机构地区：[1]青岛大学医学院,266023

出　　处：《中华实验外科杂志》2015年第7期1522-1526,共5页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81171774、81272056）

摘　　要：目的 探讨单纯转化生长因子-β3（TGF-β3）和TGF-β3与基质金属蛋白酶抑制剂-2联合转染兔骨髓间充质干细胞复合丝素蛋白/壳聚糖（SF-CS）生物支架植入动物体内修复兔膝关节软骨缺损的可行性及效果差异.方法 将新西兰大白兔20只分为4组（支架组、未转染组、reAAV-TGF-β3转染组、联合转染组）,每组5只.按组别重组腺相关病毒转染兔骨髓间充质干细胞与SF-CS生物支架复合,分别植入兔膝关节软骨缺损处.2个月后处死家兔,肉眼观察以及苏木素-伊红（HE）染色评定缺损软骨修复情况.并进行软骨细胞特征性甲苯胺蓝染色鉴定.结果 2个月后除支架组外各实验组兔膝关节软骨缺损处均有软骨样物质形成,且联合转染组诱导的新生软骨更接近缺损处周围正常软骨.Moran观察评分分别为支架组（0.600±0.548）分、未转染组（1.800±0.447）分、reAAV-TGF-β3转染组（3.800±0.837）分、联合转染组（5.400±0.548）分.联合转染组与reAAV-TGF-β3转染组间,reAAV-TGF-β3转染组与未转染组间差异均有统计学意义（P＜0.05）.HE染色结果提示联合转染组软骨修复效果较单纯TGF-β3转染组更好.结论 单纯TGF-β3转染兔骨髓间充质干细胞对兔膝关节软骨缺损有修复作用,TGF-β3与基质金属蛋白酶抑制剂-2（TIMP-2）联合转染组修复缺损效果更明显,提示TIMP-2与TGF-β3具有协同效应.Objective This study was designed to investigate the feasibility of transforming growth factor-β3 transfection and transforming growth factor-β3 （TGF-β3）,Matrix metalloproteinase inhibitors-2 co-transfection of rabbit bone marrow mesenchymal stem cells repair rabbit＇ s cartilage defect and effect differences between them,by embedded in filament fibroin （silk fibroin,SF）/chitosan （chitosan,CS） scaffolds and implanted in vivo rabitts.Methods Twenty New Zealand white rabbits were divided into four groups,five arbbirs in each group.Under sterile conditions,we took the third generation of the logarithmic growth phase of rabbit bone marrow mesenchymal stem cells （BMSCs）.And we used recombinant adeno-associated virus which carrying each group＇ s purpose gene to transfected them.After two months we killed the rabbits,as well as visually assessed of cartilage defect repair situation by hematoxylin-eosin staining （HE） staining.And conduct characteristic of cartilage cells that stained with toluidine blue staining and type Ⅱ collagen immunohistochemical staining.Results We observed that：after two months,cartilage-like substance formed in each expermental group＇ s cariliage defect area,and the co-transfection group induced cartilage were closer to hyaline cartilage.We found the Moran score of difference groups were：concrol group：0.600 ± 0.548,non-transfection group：1.800 ± 0.447,pure TGF-β3 transfected group：3.800 ± 0.837,co-transfection group：5.400 ± 0.548.And the difference of score between co-transfection group and pure TGF-β3 transfected group was statistically significant （P ＜ 0.01）.HE staining results suggested that co-transfected cartilage repair group＇ s result was better than pure TGF-β3 transfected group.Conclusion In animal experiments,pure TGF-β3 transfected rabbit bone marrow mesenchymal stem cells can repair rabbit articular cartilage defects,TGF-β3 and matrix metalloproteinases inhibitors-2 （TIMP-2） co-transfection group was more effective.

关 键 词：转化生长因子 基质金属蛋白酶抑制剂 骨髓间充质干细胞 软骨缺损 丝素蛋白/壳聚糖生物支架 

分 类 号：R781.3[医药卫生—口腔医学]