骨髓间充质干细胞定向诱导移植修复关节软骨

背景:骨髓间充质干细胞在不同诱导条件下具有向中胚层组织细胞如成骨细胞、成软骨细胞、肌细胞、脂肪细胞等分化的能力.目的:验证用组织工程方法诱导分化骨髓间充质干细胞修复兔关节软骨损伤的效果.设计、时间及地点:随机对照动物实验,于2005-05-03/2007-12-30在沈阳医学院临床中心实验室完成.材料:20只两三月龄的健康新西兰白兔,雌雄不限.方法:①诱导分化体外培养的兔骨髓间充质干细胞.实验组加入地塞米松、碱性成纤维细胞生长因子、维生素C培养1周,再将转化生长因子β替换碱性成纤维细胞生长因子培养3周;以不加诱导剂细胞做对照.②取20只兔.建立膝关节软骨缺损模型,随机分为3组.实验组10只膝关节内植入经诱导的骨髓间充质干细胞;对照组植入未经诱导的骨髓间充质干细胞:空白对照组植入生理盐水.分别于术后2,4,6,8周时处死实验组处死2只,对照组和空白对照组各处死1只进行各项指标检测.主要观察指标:①细胞形态学.②碱性磷酸酶活性的测定.③大体标本观察.④X射线观察.⑤组织切片观察.结果:①经诱导的骨髓间充质干细胞,细胞形态发生明显变化,逐渐由长梭形变为多角形,类似于软骨细胞样形态.②骨髓间充质干细胞经诱导4周后,其碱性磷酸酶活性明显增强(P＜0.05).③术后8周,实验组标本修复组织表面光滑,与周围软骨间界限模糊不清:X射线表现为关节间隙变宽,软骨下骨质囊变得到改善:组织切片观察显示与正常软骨细胞基本上一致.结论:自体间充质干细胞移植可修复关节软骨损伤.

Chondrogenic differentiation of mesenchymal stem cells for repairing articular cartilage

BACKGROUND:Under different induction conditions,bone marrow mesenchymal stem cells can differentiate into the mesodermal tissues such as osteoblasts,chondroblasts,muscle cells,adipocytes and so on.OBJECTIVE:To verify the effect on repairing the rabbit articular cartilage injury using bone marrow mesenchymal stern cells (MSCs)induced by tissue engineering method.DESIGN,TIME AND SETTING:Randomized controlled animal experiment was performed in the Clinical Center Laboratory of Shenyang Medical College between May 2005 and December 2007.MATERIALS:Twenty health New Zealand rabbits,irrespective of genders,aged 2-3 months,were used.METHODS:①Rabbit bone marrow MSCs were cultured in vitro,experiment group was cultured for one week withdexamethasone,basic fibroblast growth factor and vitamin C,then for additional 3 weeks with transforming growth factor-β instead of basic fibroblast growth factor;calls without inductors served as controls;②Twenty rabbits were used to establish knee articular cartilage defect models,which were then divided into three groups at random. Experiment group (n=5) was transplanted with the induced bone marrow MSCs;control group with the non-induced cells;blank control group with saline. At 2,4,6,8 weeks postoperation,two rabbits in the experiment group were killed,while one animal in control group and blank control group was killed for the index determination.MAIN OUTCOME MEASURES:①Cell morphology. ②Alkaline phosphatase activities.③General observation. ④X-ray observation.⑤Histological observation.RESULTS:①The morphology of the induced bone marrow MSCs was changed,from long fusiform to polygon,which was similar to cartilage calls-like morphology.②After the bone marrow MSCs were induced for 4 weeks,the alkaline phosphatase activities were obviously enhanced(P＜0.05).③Eight weeks after transplantation,the specimens in the experiment group exhibited smooth surface and unclear outlines with surrounding cartilage;X-ray results showed joint space broadened,subchondral bone sack was improved;histological slices observation revealed similarity with normal chondrocytes.CONCLUSION:Autologous MSCs transplantation can repair articular cartilage injury.