玻璃酸钠及胎盘间充质干细胞和诱导的软骨细胞膝关节腔内注射:修复膝骨关节炎  

Intra-articular injection of sodium hyaluronate versus placenta-derived mesenchymal stem cells-differentiated chondrocytes for treatment of knee osteoarthritis

作　　者：李治[1] 赵伟[1] 刘伟[2] 周晔[2] 贾京桥 杨立枫[1] 

机构地区：[1]沈阳医学院附属中心医院骨科,辽宁省沈阳市110024 [2]沈阳医学院附属中心医院产科,辽宁省沈阳市110024 [3]辽宁省维尔克干细胞生物科技有限公司,辽宁省本溪市117000

出　　处：《中国组织工程研究》2014年第50期8140-8146,共7页Chinese Journal of Tissue Engineering Research

基　　金：辽宁省科技厅科研项目(2012225014)~~

摘　　要：背景:胎盘间充质干细胞已被证实具有较强的增殖能力,能向神经细胞、血管内皮细胞、表皮细胞以及胰岛样细胞等诱导分化,但向软骨细胞诱导分化并修复膝骨关节的研究不多。目的:人胎盘间充质干细胞体外诱导软骨细胞膝关节腔内注射修复兔膝骨关节炎,并比较璃酸钠和胎盘间充质干细胞膝关节腔内注射的修复效果。方法:1选新西兰大白兔制作骨关节炎模型,左后肢膝关节炎造模后石膏固定5.5周,镜下观察关节软骨情况。2利用Ⅳ型胶原酶消化分离培养人胎盘间充质干细胞,流式细胞仪检测细胞表面标志物,证实为胎盘间充质干细胞。取第3代胎盘间充质干细胞进行实验,将胎盘间充质干细胞加入软骨诱导培养基向软骨细胞分化,倒置显微镜观察结果。3将大白兔随机分成玻璃酸钠膝关节腔内注射组、胎盘来源间充质干细胞膝关节腔内注射组和诱导软骨细胞膝关节腔内注射组,分别给予膝关节腔内注射,连续5周取膝关节软骨镜下观察,比较软骨修复情况。结果与结论:体外诱导培养的人胎盘间充质干细胞增殖形成大量贴壁细胞,贴壁细胞具有典型的间充质细胞形态;应用软骨诱导培养基培养后,细胞只保持微团,不继续增殖形成贴壁细胞,微团基部无贴壁细胞,显示人胎盘间充质干细胞可以分化为软骨细胞。苏木精-伊红染色镜下观察发现,各组兔膝关节软骨细胞均有修复,诱导软骨细胞膝关节腔内注射后兔膝关节软骨修复情况较好,优于玻璃酸钠和胎盘间充质干细胞膝关节腔内注射组。BACKGROUND： Placenta-derived mesenchymal stem cells have been shown to have a strong proliferative capacity, which can be induced to differentiate into nerve cells, vascular endothelial cells, epithelial cells and islet-like cells, but there are few studies about the chondrogenic differentiation in the treatment of knee osteoarthritis. OBJECTIVE： To compare the intra-articular injection of sodium hyaluronate and placenta-derived mesenchyma stem cells-induced chondrocytes for treatment of knee osteoarthritis. METHODS： （1） Knee osteoarthritis models were established in the left hind of New Zealand white rabbits. After 5.5 weeks of plaster fixation, the articular cartilage was observed under microscope. （2） Human placanta-derived mesenchymal stem cells were isolated and cultured using IV collagenase digestion, and flow cytometry was used to detect cell surface markers to identify the placenta-derived mesenchymal stem cells. Passage 3 placenta-derived mesenchymal stem cells were cultured in chondrocyte induction medium and observed under inverted microscope. （3） The rabbits were randomly divided into sodium hyaluronate group, placenta-derived mesenchymal stem cells group, and induced chondrocyte group, and then intra-articular injection of sodium hyaluronate, placenta-derived mesenchymal stem cells, and induced chondrocyte was given, respectively. After 1-5 weeks, the knee cartilage was taken and observed under microscope. RESULTS AND CONCLUSION： In vitro cultured human placenta-derived mesenchymal stem cells proliferated to produce a large amount of adherent cells with typical mesenchymal cell morphology; after culture in inducing culture medium, cells presented with micelles and could not proliferate to form adherent cells. There were no adherent cells on the micelles base, showing human placenta-derived mesenchymal stem cells can differentiate into chondrocytes. Under hematoxylin-eosin staining, rabbit articular cartilage was repaired in all the groups, but the intra-articular injection of chon

关 键 词：干细胞 间质干细胞 软骨细胞 骨关节炎 膝 移植 胎盘间充质干细胞 玻璃酸钠 膝骨关节 动物模型 细胞培养 体外诱导 苏木精-伊红染色 胶原酶 细胞微团 

分 类 号：R394.2[医药卫生—医学遗传学]