人脱细胞羊膜与骨髓间充质干细胞复合体修复关节软骨缺损  

Repair of articular cartilage defects with human acellular amniotic membrane/bone marrow mesenchymal stem cell composite

作　　者：姜良斌[1,2] 韦标方 冯志[2] 岳永彬[1,2] 

机构地区：[1]广州中医药大学,广东省广州市510006 [2]临沂市人民医院,山东省临沂市276000

出　　处：《中国组织工程研究》2017年第26期4113-4118,共6页Chinese Journal of Tissue Engineering Research

基　　金：山东省科技发展计划(2014GSF119022)~~

摘　　要：背景:骨髓间充质干细胞在软骨组织工程研究中作为一种常见的种子细胞被广泛应用于软骨缺损修复。目的:以人脱细胞羊膜作为细胞支架负载兔骨髓间充质干细胞用以修复兔股骨髁间窝软骨缺损。方法:将兔骨髓间充质干细胞接种于人脱细胞羊膜上,体外共培养2周。建立兔股骨髁间窝区关节软骨缺损模型,右膝软骨缺损处设为空白对照组,不植入任何材料,左膝软骨缺损处为实验侧,分别植入人脱细胞羊膜-骨髓间充质干细胞和单纯人脱细胞羊膜。术后第8,12周,取膝关节软骨缺损部位新生组织,行大体观察、组织学检测,评估新生软骨质量。结果与结论:(1)大体观察结果:人脱细胞羊膜-骨髓间充质干细胞组有类软骨形成,质地较软,与周围正常软骨结合良好;人脱细胞羊膜组未形成软骨组织。空白对照组缺损区只由纤维样组织填充;(2)组织学观察结果:人脱细胞羊膜-骨髓间充质干细胞组新生出软骨细胞及软骨陷窝,并形成软骨基质,Ⅱ型胶原免疫组织化学染色呈阳性。软骨基质甲苯胺蓝染色较深。人脱细胞羊膜组仅有极少量的软骨细胞,甲苯胺蓝染色较浅,Ⅱ型胶原免疫组织化学染色呈阴性,无软骨基质形成。空白对照组为纤维组织修复,甲苯胺蓝染色较浅;(3)结果表明,人脱细胞羊膜有利于骨髓间充质干细胞的软骨化,软骨缺损处由类软骨组织填充,能够有效修复关节软骨缺损。BACKGROUND： Bone marrow mesenchymal stem cells （BMSCs） as common seed cells have been widely used in tissue-engineered cartilage repair.OBJECTIVE： To use human amniotic membrane as a cell scaffold to carry rabbit BMSCs in order to repair articular cartilage defects in the femoral intercondylar fossa of rabbits.METHODS： Rabbit BMSCs were inoculated onto the human acellular amniotic membrane （HAAM） and co-cultured for 2 weeks. Articular defect models were made in the femoral intercondylar fossa of rabbits. The defects of the right knees served as blank control. BMSCs/HAAM composite was transplanted into the defect of the left knee joint as composite group, and HAAM was implanted into the defect of the left knee joint as HAAM group. These rabbits were killed at 8 and 12 weeks after implantation and the newly cartilage samples were evaluated grossly and histologically and then graded.RESULTS AND CONCLUSION： Gross observation showed the defects were filled with cartilaginous tissues in the composite group, and there were no cartilage tissues in the HAAM group, while only fibrous tissues were seen in the blank control group. Histologically, the defect region was full of chondrocytes in the composite group,immunohistochemistry staining indicated that collagen II was rich in the tissue, and furthermore, the cartilage matrix was stained deeply by toluidine blue. In the the HAAM group, there were few chondrocytes, toluidine blue staining was weakly positive, and immunohistochemistry staining was negative, indicating there was no cartilage matrix. In the blank control group, the defects were filled of fibroblasts and toluidine blue staining was weakly positive. To conclude, the BMSCs/HAAM is a good scaffold for BMSCs chondrogenic differentiation to effectively repair articular cartilage defects.

关 键 词：生物材料 软骨生物材料 羊膜 骨髓间充质干细胞 软骨缺损 兔 

分 类 号：R318[医药卫生—生物医学工程]