兔成骨细胞在羟基丁酸-羟基辛酸一体化骨软骨支架上的亲和性研究  

Study on the affinity of rabbit osteoblasts in poly（hydroxybutyrate-co-hydroxyoctanoate） osteochondral scaffold

作　　者：张烨[1] 张永红[1] 余兆泽 刘旻[1] 赵良启[2] 

机构地区：[1]山西医科大学第二医院骨科,太原030001 [2]山西大学生物技术研究所

出　　处：《中国临床实用医学》2015年第2期40-43,共4页China Clinical Practical Medicine

基　　金：国家自然科学基金项目（31040030）

摘　　要：目的：成骨化诱导兔骨髓间充质干细胞（BMSCs），并将诱导的细胞与羟基丁酸-羟基辛酸新型材料聚合物一体化骨软骨支架结合在一起培养，观察该细胞在支架上的增殖和分化情况。方法2014年6至12月，于山西大学化学生物学与分子工程实验室，运用羟基丁酸-羟基辛酸聚合物新型材料制备具有骨层-中间界面层-软骨层结构的仿生一体化骨软骨支架。将传代培养至第3代的兔 BMSCs 放入成骨细胞诱导液中诱导18 d，将诱导的成骨细胞接种到一体化骨软骨支架上，MTT 法检测细胞在支架上的增殖活性，7d 后电镜观察细胞在支架内的生长状况。结果新型材料制备的仿生一体化支架具有3层结构，不但结构连接紧密，而且支架孔隙均匀而且相互交通呈立体网状分布，支架孔隙率为74%，利于支架同一层内细胞的依附增殖和迁移。结论细胞与支架结合培养7 d 后，电镜下可见细胞依附在骨层支架孔隙内及周边，生长增殖状态良好，并未进入支架中间层。Objective Between rabbit bone marrow mesenchymal cells induced into osteoblasts and co-cultured with poly（hydroxybutyrate-co-hydroxyoctanoate） osteochondral scaffold, to observe cells and scaffold affinity. Methods From June to December 2014,The poly（hydroxybutyrate-co-hydroxyoctanoate） osteochondral scaffold having a three-layer structure （layer of bone/bone and cartilage interface layer/layer of cartilage） was prepared by solvent casting/particle leaching method in the Chemical Biology and Molecular Engineering Laboratory of Shanxi University. The subcultured to the third generation of the rabbit bone marrow mesenchymal stem cells into osteoblasts induced by fluid were induced for 18 days .The induced bone marrow mesenchymal stem cells at passage 3 were seeded onto the scaffold of bone layer. The proliferation of the osteoblasts growing on the scaffolds was examined using 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide method, the growth of cells in the scaffold was observed under electron microscope after 7 days. Results The integrated osteochondral scaffold had a clear appearance of three-layer structure, which had closed connections between the three layers. Porous bone layer was visible as well as uniform and interlinked pores, and the porosity was 74%. The osteoblasts seeded onto the scaffold proliferated well and presented a three-dimensional growth after 7 days of co-culture, but there were no cells on the interface layer. Conclusion Cells showed a luminal-like structure growing on the scaffold with the porous structure, but they did not grow into the interface layer of bone and cartilage.

关 键 词：羟基丁酸 - 羟基辛酸共聚体 支架 组织工程 成骨细胞 亲和性 

分 类 号：R764.91[医药卫生—耳鼻咽喉科]