两种可注射Pluronic F-127水凝胶复合物修复兔软骨缺损的效果差异  

Differences in the effect of two injectable Pluronic F-127 hydrogel composites in repairing rabbit cartilage defects

作　　者：樊薰勤 张明勇[1] 戴能 王喆敏 刘拴[1] Fan Xunqin;Zhang Mingyong;Dai Neng;Wang Zhemin;Liu Shuan(Department of Orthopedics,Tianyou Hospital Affiliated to Wuhan University of Science and Technology,Wuhan 430064,Hubei Province,China)

机构地区：[1]武汉科技大学附属天佑医院骨科

出　　处：《中国组织工程研究》2020年第16期2506-2512,共7页Chinese Journal of Tissue Engineering Research

基　　金：湖北省卫计委项目(WJ2017M171)，项目名称：SOX-9基因修饰的骨髓间充质干细胞在促进骨肌腱结合部位愈合的相关机理研究，项目负责人：刘拴~~

摘　　要：背景:预实验显示,SOX9基因转染的骨髓间充质干细胞可在PluronicF-127水凝胶内良好的生长与增殖,促进细胞外基质的分泌,增加软骨基质的表达。目的:利用慢病毒基因诱导方式将SOX9基因转导至骨髓间充质干细胞中,将其与可注射Pluronic F-127水凝胶复合,观察Pluronic F-127水凝胶复合物修复软骨缺损的效果。方法:利用慢病毒基因诱导方式将SOX9基因转导至骨髓间充质干细胞中,转染48 h后与Pluronic F-127水凝胶复合。取60只新西兰大白兔(武汉科技大学实验动物中心提供),建立右侧膝关节股骨髁软骨缺损模型,随机分3组处理:模型组缺损部位未植入任何材料,对照组植入未转染的骨髓间充质干细胞与Pluronic F-127水凝胶复合物,实验组植入SOX9基因转染的骨髓间充质干细胞与PluronicF-127水凝胶复合物。术后4,12周取缺损部位组织,分别进行Micro-CT三维重建、苏木精-伊红染色、番红O染色、Ⅱ型胶原免疫组织化学染色与Wakitani软组织损伤修复组织学评分。实验获得武汉科技大学伦理委员会批准。结果与结论:①术后12周Micro-CT三维重建显示,模型组缺损区域未见明显的修复,中央仍有较大的凹陷;对照组可见明显的修复,中央凹陷区域明显减小,可见较多的再生骨小梁结构;实验组缺损部位基本完成修复;②术后12周苏木精-伊红染色显示,模型组缺损区仍未见骨小梁结构,细胞分布紊乱,未见软骨陷窝;对照组可见较多的骨组织重建,缺损区域主要由软骨样组织与纤维组织填充;实验组骨组织重建较充分,缺损区域主要由软骨样细胞与软骨样细胞外基质填充,细胞呈柱状排列,与周围软骨相似;③术后12周番红O染色与Ⅱ型胶原免疫组织化学染色显示,模型组可见少量糖胺多糖表达,未见Ⅱ型胶原表达;对照组可见较多的糖胺多糖与Ⅱ型胶原表达,实验组糖胺多糖与Ⅱ型胶原表达最多;④实验组Wakitani软BACKGROUND: Preliminary experiments show that bone marrow mesenchymal stem cells transfected with SO X9 gene can grow and proliferate in Pluronic F-127 hydrogel, promote the secretion of extracellular matrix, and increase the expression of cartilage matrix. OBJECTIVE: The SOX9 gene was transduced into bone marrow mesenchymal stem cells by lentivirus gene induction, and then combined with injectable Pluronic F-127 hydrogel to observe the effect of Pluronic F-127 hydrogel on repairing cartilage defects. METHODS: SOX9 gene was transfected into bone marrow mesenchymal stem cells by lentivirus gene induction. After 48 hours of transfection, SOX9 gene was combined with Pluronic F-127 hydrogel. Sixty New Zealand white rabbits provided by the Experimental Animal Center of Wuhan University of Science and Technology were selected to establish the models of femoral condylar cartilage defect of the right knee joint. The rabbits were randomly divided into three groups: model group without implantation of any material at the defect site, control group with implantation of non-transfected bone marrow mesenchymal stem cells and Pluronic F-127 hydrogel complex at the defect site, and experimental group with implantation of SOX9 gene-transfected bone marrow mesenchymal stem cells and Pluronic F-127 hydrogel complex at the defect site. Four and twelve weeks after operation, the defect tissues were taken for three-dimensional reconstruction of micro-CT, hematoxylin-eosin staining, Safranine O staining, type II collagen immunohistochemical staining and Wakitani soft tissue repair histological score. This study was approved by the Ethics Committee of Wuhan University of Science and Technology. RESULTS AND CONCLUSION:(1) At 12 weeks after operation, three-dimensional reconstruction of Micro-CT showed that there was no obvious repair in the defect area of the model group, and there was still a large depression in the center. In the control group, the central depression area was significantly reduced and more trabecular structures of reg

关 键 词：Pluronic F-127水凝胶 SOX9基因 软骨缺损 Ⅱ型胶原 骨髓间充质干细胞 水凝胶复合物 糖胺多糖 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学]