大鼠骨髓间充质干细胞与多孔双相磷酸钙陶瓷支架体外黏附的实验研究  

EXPERIMENTAL STUDY ON CELL ADHESION CHARACTERISTIC BETWEEN POROUS BIPHASIC CALCIUM PHOSPHATE NANOCOMPOSITE AND BONE MARROW MESENCHYMAL STEM CELLS IN VITRO

作　　者：王涛[1] 田卫东[2] 陈希哲[2] 李声伟[2] 廖运茂[2] 

机构地区：[1]海南省人民医院口腔科,海口570003 [2]四川大学口腔生物医学工程教育部重点实验室,成都610041

出　　处：《中国修复重建外科杂志》2006年第5期565-568,共4页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：四川省科技厅科技攻关资助项目(03GG009-027;03SG022-003-1);国家教育部优秀青年教师教学科研奖励计划资助项目(2003682)~~

摘　　要：目的研究大鼠骨髓间充质干细胞(marrowmesenchymalstemcells,MSCs)诱导培养后在多孔双相磷酸钙(biphasiccalciumphosphate,BCP)陶瓷支架材料上的黏附和生长。方法SD大鼠MSCs经矿化诱导培养、扩增,具有成骨细胞表型后,与多孔BCP陶瓷支架及普通多孔羟基磷灰石(hydroxyapatite,HA)陶瓷支架体外复合培养,扫描电镜观察比较MSCs在两种材料支架表面的黏附数量和形态;同时以0.5、1.0、2.0、3.0和4.0×106/ml浓度细胞悬液接种于多孔BCP支架材料,检测适宜的接种浓度及单位体积支架材料可黏附MSCs数量。结果大鼠MSCs经诱导培养14d后,行矿化沉积茜素红染色、型胶原免疫细胞化学染色及碱性磷酸酶细胞化学染色,结果均为阳性。大鼠MSCs黏附于多孔BCP陶瓷上的细胞数(88.00±6.58)明显高于HA陶瓷组(39.00±3.65),两组比较差异有统计学意义(P<0.01)。当接种浓度为2.0×106/ml时,单位体积的陶瓷支架材料最多可黏附MSCs数量为1.28×107个/cm3,为细胞适宜接种浓度。结论大鼠MSCs在体外经矿化诱导培养可表达成骨细胞表型,细胞浓度为2.0×106/ml时与多孔BCP陶瓷支架材料具有良好的黏附能力。Objective To study the adhesion characteristic in vitro between porous biphasic calcium phosphate （BCP） nanocomposite and bone marrow mesenchymal stem cells （MSCs） that have been induced and proliferated. Methods MSCs obtained from SD rat bone marrow were in vitro induced and proliferated. After their osteoblastic phenotype were demonstrated, MSCs were seeded onto prepared porous BCP nanocomposite （experiment group）and common porous hydroxyapatite （control group ）. Their adhesion situation was analyzed by scanning electron microscope. The initial optimal cell seeding density was investigated between new pattern porous BCP nanocomposite and MSCs by MTT automated colormetric microassay method. Results The differentiation of MSCs to osteoblastic phenotype were demonstrated by the positive staining of mineralized node, alkaline phosphatase （ALP） and collagen type Ⅰ , the most appropriate seeding density between them was 2×10^6/ml. The maximal number which MSCs could adhere to porous BCP nanocomposite was 1. 28×10^7/cm^3. Conclusion MSCs can differentiate to osteoblastic phenotype. The MSCs were well adhered to porous BCP nanocomposite.

关 键 词：组织工程骨 骨髓间充质干细胞 多孔双相磷酸钙陶瓷支架 大鼠 

分 类 号：R318.08[医药卫生—生物医学工程] Q813[医药卫生—基础医学]