异体滑膜及青少年软骨颗粒移植修复兔半月板“白区”缺损及移植细胞的转归研究  

The Fate of Donor Cells in Synovium and Particulated Juvenile Allograft Cartilage for the Repair of Meniscal “white zone” Defects

作　　者：闫文强 吴桐 樊逸菲 麦合木提·麦麦提敏 赵逢源 李豫皖 曹宸喜 胡晓青[1] 程锦[1] 敖英芳[1] Yan Wenqiang;Wu Tong;Fan Yifei;Maihemuti Maimaitimin;Zhao Fengyuan;Li Yuwan;Cao Chenxi;Hu Xiaoqing;Cheng Jin;Ao Yingfang(Department of Sports Medicine,Peking University Third Hospital,Institute of Sports Medicine of Peking University,Beijing Key Laboratory of Sports Injuries,Beijing 100191,China)

机构地区：[1]北京大学第三医院运动医学科北京大学运动医学研究所运动医学关节伤病北京市重点实验室,北京100191

出　　处：《中国运动医学杂志》2022年第12期941-955,共15页Chinese Journal of Sports Medicine

基　　金：国家自然科学基金(82172420,81972101,81802161,81871770)。

摘　　要：目的:探讨异体滑膜和青少年软骨颗粒移植在半月板“白区”缺损修复上的协同作用;研究移植细胞的转归(存活和分化);研究滑膜间充质干细胞和软骨细胞间的相互作用。方法:选取2只未成年雄性兔作为滑膜及青少年软骨颗粒供体。选取24只成年雌性兔(共48个膝关节)制备半月板缺损模型,分为以下几组:空白组,半月板缺损无任何处理;单纯纤维蛋白胶组,在半月板缺损处填充单纯纤维蛋白胶;纤维蛋白胶+滑膜移植组,纤维蛋白胶包裹异体滑膜碎片填充到半月板缺损处;纤维蛋白胶+滑膜+异体青少年软骨颗粒移植组,纤维蛋白胶包裹异体滑膜和异体青少年软骨颗粒填充到半月板缺损处,每组样本量为12个膝关节。分别在术后8周和16周取材,通过组织学和免疫组织化学染色评估半月板缺损修复程度,针对雄性个体特有的Y染色体性别决定区基因(SRY)进行核酸原位杂交,并结合免疫荧光共定位评估移植细胞的存活和分化。其次,建立体外兔滑膜间充质干细胞和软骨细胞共培养系统,检测半月板纤维软骨细胞相关基因转录水平,并探索细胞间相互作用的可能机制。结果:在术后8、16周,相较于空白组、单纯纤维蛋白胶组及纤维蛋白胶+滑膜移植组,纤维蛋白胶+滑膜+异体青少年软骨颗粒移植组的缺损表现出良好的结构完整性、表面光滑度和边缘整合度,修复组织中可见较多的半月板样纤维软骨细胞,并且,细胞周围的基质在甲苯胺蓝和II型胶原免疫组织化学染色中表现为强阳性。在术后16周,纤维蛋白胶+滑膜移植组及纤维蛋白胶+滑膜+异体青少年软骨颗粒移植组的SRY核酸原位杂交信号表现为强阳性,表明移植细胞在受体中至少可存活16周。其次,核酸原位杂交和SOX9免疫荧光共定位显示表面移植滑膜组织中的细胞表现为SOX9阳性,表明移植滑膜组织中的细胞可分化为半月板纤维软骨�Objective To explore the synergistic effect of synovium and particulated juvenile allograft cartilage(PJAC) on repairing meniscal defects in the avascular zone,track the survival and differentiation of donor cells and interactions between synovial cells and chondrocytes.Methods In this study,2immature male rabbits were raised as synovium and particulated cartilage donors.Meanwhile,24 adult female rabbits were selected and induced cylindrical full-thickness defects measuring 2.0 mm in the avascular portion of medial menisci in both knees.They were then randomly divided into a blank group,a fibrin gel group,a fibrin gel+synovium transplant group as well as a fibrin gel+synovium and PJAC transplant group,each of 12 rabbits.The blank group was left untreated,while the fibrin gel group,fibrin gel+synovium transplant group as well as fibrin gel+synovium and PJAC transplant group were filled into the meniscal defects with fibrin gel,fibrin gel encapsulating synovium fragments,as well as fibrin gel encapsulating synovium fragments and PJAC respectively.Rabbits were sacrificed 8 and 16 weeks postoperatively,and the histological and immunohistological methods were applied to assess the degree of meniscal repair.The in situ hybridization(ISH) for male-specific sex-determining region Y-linked(SRY) gene and immunofluorescence for Sry-type HMG-box 9(SOX9) were conducted to evaluate the survival and differentiation of donor cells.Moreover,in vitro cell coculture systemofsynovium derived mesenchymal stem cells(SMSCs) and chondrocytes was built,and their interaction was analyzed.Results Eight and 16 weeks after the operation,better structural integrity,superficial smoothness and marginal integration were observed in the fibrin gel+synovium and PJAC transplant group,compared with the other 3 groups,with more meniscus-like fibrochondrocytes in the repaired tissue,and the matrix around such cell clusters being strongly positive after the toluidine blue and type 2 collagen immunohistochemistry staining.The SRY signals of ISH were pos

关 键 词：半月板损伤 半月板修复 滑膜 异体青少年软骨颗粒 

分 类 号：R684[医药卫生—骨科学]