兔骨髓基质细胞体外成骨分化鉴定及与煅烧骨复合培养的实验研究  

The osteoblast differentiation of rabbit bone marrow stromal cells and construction with true bone ceramic

作　　者：杨成林[1] 毕郑钢[1] 王玉学[1] 祁权[1] 付春江[1] 陶天遵[2] 

机构地区：[1]哈尔滨医科大学附属第一医院骨科,150001 [2]哈尔滨医科大学附属第二医院骨科

出　　处：《中国骨质疏松杂志》2005年第3期329-332,共4页Chinese Journal of Osteoporosis

摘　　要：目的研究兔骨髓基质细胞体外向成骨细胞分化的生物学特性及其与牛煅烧骨体外复合培养的生物相容性。方法将体外培养1周的兔骨髓基质细胞向成骨细胞诱导分化,于1、2、4周时提取RNA,采用RTPCR技术检测ALP和I型胶原mRNA表达,并对培养2周的细胞行VonKossa染色观察钙结节形成;另制备细胞煅烧骨复合物,继续培养1、7、14d后取出,扫描电镜观察及X射线衍射仪检测。结果体外诱导培养2、4周后,兔骨髓基质细胞成功表达ALT和I型胶原,VonKossa染色可见钙结节形成。扫描电镜及X射线衍射分析证实细胞在煅烧骨表面大量贴附成活,14d后细胞铺满支架表面,合成并分泌胶原纤维及钙盐。结论兔骨髓基质细胞体外可成功向成骨细胞诱导分化,成骨特性表达佳;与牛煅烧骨体外复合培养显示良好的生物相容性。Objective To study the biological characteristics and the osteoblast differentiation ability of rabbit bone marrow stromul cells （BMSCs） cultured in experimental culture medium, and then to investigate the biocompatibility of true bone ceramic （TBC） with BMSCs cultured in vitro. Methods Rabbit BMSCs cultured for 1 week /n vitro were induced to osteoblast, to extract RNA, alkaline phosphatase（ALP） and type 1 Collagen（Col-Ⅰ） mRNA expression were measured using RT-PCR technique at different time. And mineralization were observed by VonKossa staining.Then induced BMSCs were seeded into TBC to form cell-scaffold construct,and the cell-scaffold construct cultured continually in the same culture medium were examined with scaning electronic microscopy （SEM）, Results In vitro induced BMSCs exhibited an osteogenic phynotype. After 2 weeks, ALP and type 1 Collagen were expressed in the experimental BMSCs, and calcium nodules were observed by VonKossa staining. And 1^st .7^th day ,a lot of cells grew in and on TBC, 14^th day, cells grew all over scaffold, collagenic fibers and calcic granules was found on the cell-scaffold construct. Conclusions In vitro rabbit BMSCs can be successfully induced to osteoblast, and expressing osteogenic characteristics significantly. After prepared cells-scaffold composite, TBC have good compatibihty with induced BMSCs.

关 键 词：骨髓基质细胞 成骨细胞 煅烧骨 生物相容性 骨髓基质细胞 体外复合培养 牛煅烧骨 体外成骨 分化鉴定 PCR技术检测 实验研 细胞诱导分化 mRNA表达 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R-332[医药卫生—基础医学]