TGF-β3与TIMP-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(英文)  

TGF-β_3 and TIMP-2 Co-transfection of Rabbit Bone Marrow Mesenchymal Stem Cells Compound Silk Fibroin / Chitosan Scaffolds Implanted in Vivo Rabbit Cartilage Defect Repair; Silk Fibrin/Chitosan Biological Scaffold

作　　者：孟飞[1] 吕成昱[2] 张海宁[2] 隋爱华[2] 刘迎[1] 

机构地区：[1]青岛大学医学院,山东青岛266021 [2]青岛大学医学院附属医院,山东青岛266021

出　　处：《现代生物医学进展》2014年第14期2622-2627,共6页Progress in Modern Biomedicine

基　　金：National Natural Science Foundation project funding(81171774;81272056)~~

摘　　要：目的:TGF-β3广泛存在于骨组织、软骨组织中,能诱导体外培养的间充质干细胞向软骨分化、生长。TIMP-2能抑制MMP对软骨基质的降解,保护新生软骨组织。本实验探讨单纯TGF-β3和TGF-β3,TIMP-2联合转染兔骨髓间充质干细胞复合丝素蛋白壳聚糖/(silk fibrin/chitosan,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,每组5只(支架组、未转染组、reAAV-TGF-β3转染组、reAAV-TGF-β3,reAAV-TIMP-2联合转染组)。在无菌条件下取兔第三代对数生长期骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs),用携带目的基因的重组腺相关病毒进行转染,将转染成功的BMSCs与SF-CS生物支架复合,分别植入兔膝关节软骨缺损处:未转染组植入SF-CS生物支架,未转染组植入未转染的BMSCs复合SF-CS生物支架,reAAV-TGF-β3转染组植入reAAV-TGF-β3转染的BMSCs复合SF-CS生物支架,reAAV-TGF-β3,TIMP-2联合转染组植入reAAV-TGF-β3,TIMP-2联合转染的BMSCs复合SF-CS生物支架。两月后处死家兔,肉眼观察以及HE染色评定缺损软骨修复情况。并进行软骨细胞特征性染色即甲苯胺蓝染色及II型胶原免疫组化染色鉴定。结果:两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成分更接近缺损处周围正常软骨。联合转染组与reAAV-TGF-β3转染组;联合转染组与未转染组;reAAV-TGF-β3转染组与未转染组的评分差异均具有统计学意义(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： TGF-β3widely present in bone tissue and cartilage tissue, can induce cultured mesenchymal stem cells into cartilage differentiation and growth. TIMP-2 can inhibits the degradation of the cartilage matrix caused by MMP and protect new cartilage. This test investigated the feasibility and effect differences of pure TGF-β3and TGF-β3, TIMP-2 co-transfection of rabbit bone marrow mesenchymal stem cells Filament fibroin（silk fibroin, SF） / chitosan（chitosan, CS） scaffolds implanted animals repair of rabbit articular cartilage defects in vivo. Methods： Twenty New Zealand white rabbits were divided into four groups, each five（scaffold group, untransfected group, reAAV-TGF-β3transfected group, reAAV-TGF-β3, reAAV-TIMP-2 co-transfection group）. Under sterile conditions the third generation of the logarithmic growth phase of rabbit bone marrow mesenchymal stem cells（BMSCs） were took, with the purpose of carrying a recombinant adeno-associated virus gene transfection, the composite with transfected BMSCs and SF-CS scaffolds were implanted in rabbit articular cartilage defects： blank control group was implanted SF-CS scaffold, the negative control group was implanted untransfected BMSCs composite SF-CS scaffold, reAAV-TGF-β3turn transfection group implanted reAAV-TGF-β3 transfected BMSCs composite SF-CS scaffold, reAAV-TGF-β3, TIMP-2 co-transfection group was implanted reAAV-TGF-β3, TIMP-2 transfected BMSCs joint compound SF-CS scaffold. After two months, the rabbits were killed, as well as visually assessed of cartilage defect repair situation by HE staining, and conduct characteristic of cartilage cells that stained with toluidine blue staining and type II collagen immunohistochemical staining. Results： After two months, cartilage defects in each experimental group were formed cartilage-like substance, and the co-transfection group induced cartilage were closer to hyaline cartilage, the difference of co-transfection group and pure TGF-β3transfected group score was statistically s

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 

分 类 号：Q95-3[生物学—动物学] R68[医药卫生—骨科学]