异种去蛋白松质骨复合骨髓间充质干细胞修复骨缺损的组织学变化

背景：目前应用各种人造支架复合细胞修复骨缺损研究很多,但是各种人造支架都没有骨的天然结构,所以修复效果不够理想。 目的：将大鼠骨髓间充质干细胞接种到异种去蛋白松质骨上,移植修复大鼠股骨节段性缺损,以体内修复效果来评价复合体应用前景。 方法：分离培养大鼠骨髓间充质干细胞并进行扩增,用BrdU体外进行标记。同时制备牛去蛋白松质骨,在体外与标记后的细胞复合。制备大鼠双侧股骨中段5 mm缺损模型,实验分成3组,缺损处分别移植骨髓间充质干细胞/去蛋白松质骨复合体、单纯去蛋白松质骨及单纯骨髓间充质干细胞。 结果与结论：BrdU免疫染色结果显示,在各组细胞均呈阳性表达,但随着移植时间的延长而减弱。X射线放射学评分及苏木精-伊红染色组织学评分结果显示,各时间段复合体组成骨效果均好于其他组。复合体组Ⅰ型胶原蛋白表达随着时间的延长有明显增强,强于其他组。提示骨髓间充质干细胞复合异种去蛋白松质骨的成骨能力明显强于单纯的支架修复能力,单纯的骨髓间充质干细胞虽然有成骨能力,但不能修复节段性骨缺损。

Histological changes of bone marrow mesenchymal stem cells combined with heterogeneous deproteinized cancellous bone in repair of bone defects

BACKGROUND:Nowadays, there are so many studies on manmade scaffold combined with cells for repair of bone defects. However, the restoration effects are not ideal because of the manmade scaffold lacking the nature structure of bone. OBJECTIVE:To inoculate the bone marrow mesenchymal stem cells (BMSCs) in the deproteinized cancellous bone in order to repair segmental femoral defects of rats and evaluate the composite application prospect by restoration effects in vivo. METHODS: BMSCs were isolated, cultured, proliferated and marked by BrdU in vitro. At the same time bovine deproteinized cancellous bone was prepared, and then combined with the marked cells in vitro. The bone defect models (5 mm in length) were created at the bilateral femur in rats and divided into three groups. The defects were implanted by BMSCs combined with deproteinized cancellous bone, deproteinized cancellous bone, and BMSCs respectively. RESULTS AND CONCLUSION:The BrdU expressed in all cells of each group, and the expression was decreased as time went on. X-ray scores and hematoxylin-eosin stained histological scores showed that the osteogenic effect of the composite group was better than those of the other groups. Collagen protein-Ⅰ expression in the composite group was obviously increased as time went on and was stronger than the other groups. The osteogenesis ability of the composite of BMSCs and heterogeneous deproteinized cancellous bone is stronger than simple scaffolds. BMSCs have the osteogenesis ability, but they cannot repair segmental bone defects.