RCCS中犬成骨细胞-nBGC复合物联合培养的实验研究  

An Experimental Study on Canine Osteoblasts Coculture with nBGC Scaffold in RCCS

作　　者：何川[1] 邓廉夫[1] 周来生[2] 冯伟[1] 万超[1] 杨庆铭[1] 

机构地区：[1]上海第二医科大学瑞金医院骨科上海市伤骨科研究所,上海200025 [2]美国波士顿大学Goodman牙科学院

出　　处：《上海第二医科大学学报》2003年第4期301-304,共4页Acta Universitatis Medicinalis Secondae Shanghai

基　　金：国家自然科学基金 (30 0 70 759);上海市曙光基金 (2 0 0 0SG42 )资助课题

摘　　要：目的探讨联合应用成骨细胞、活性玻璃 /胶原纳米复合材料 (nBGC)和旋转式细胞培养系统 (RCCS)体外构建组织工程化骨的可行性。　方法采用骨片组织培养法 ,分离培养犬皮质骨成骨细胞 ,传代培养至第 2代后 ,利用浸泡接种方法 ,形成成骨细胞 -nBGC支架复合物 ,然后在RCCS中培养。扫描电镜观察和上清液分析了解nBGC支架上成骨细胞生长、胞外基质沉积和成骨表型维持的情况。　结果成骨细胞吸附于nBGC支架表面 ,表面可见颗粒状分泌物和丝状胶原纤维 ,形成三维结构的细胞外基质 ;上清液生化分析显示 ,成骨细胞分泌大量Ⅰ型胶原、骨特异性碱性磷酸酶 (B -AKP)和骨钙素 (OCN) ,且分别在 18、2 4和 30d达分泌高峰。　结论nBGC材料和RCCS可以为成骨细胞提供良好的生物学环境 。Objective To explore the possibility of construction of the tissue engineered bone in the rotary cell culture system (RCCS) using nano-bioactive glass/collagen (nBGC) scaffold and osteoblasts. Methods Osteoblasts were isolated and cultured from the canine cortical bone chips, and confirmed by morphology and histochemistry observation. Cells were harvested, loaded onto the porous nBGC scaffolds, and then were cultured in two vessels of RCCS to study living cell adhesion and osteogenic proteins secretion by electron microscopy and biochemistry observation. Results Electron microscopic observation revealed that the osteoblasts proliferated, produced collagen fibrils and formed 3-dimensional extracellular matrix on the scaffolds in RCCS. Biochemistry analysis showed that type Ⅰ collagen, B-AKP and OCN were present in the supernatant and their levels reached the highest point on 18, 24, 30d, respectively. Conclusion nBGC material and RCCS provided suitable microenvironment for osteoblasts. This suggests that the osteoblasts coculture with nBGC in RCCS might be an ideal method for construction of tissue engineered bone in vitro.

关 键 词：旋转细胞培养系统 活性玻璃/胶原纳米复合材料 成骨细胞 骨组织工程 

分 类 号：R318.08[医药卫生—生物医学工程]