骨髓间充质干细胞复合羟基磷灰石/磷酸三钙植骨材料的体外研究  

IN VITRO STUDIES OF BONE MARROW MESENCHYMAL STEM CELLS ON HA/TCP BONE GRAFT SCAFFOLD

作　　者：王健[1] 张海燕[1] 胡勇[2] 卢玉仙[1] 夏春林[1] 

机构地区：[1]苏州大学医学院解剖学教研室暨博习临床解剖学研究所,苏州215123 [2]江苏省建湖县人民医院骨科,建湖224700

出　　处：《解剖学报》2008年第4期539-542,共4页Acta Anatomica Sinica

基　　金：江苏省卫生厅科研基金项目(H200558)

摘　　要：目的研究兔骨髓间充质干细胞(BMSCs)在羟基磷灰石/磷酸三钙(HA/TCP)植骨材料上的黏附增殖情况。方法抽取兔股骨骨髓,进行贴壁培养BMSCs。在成骨诱导液中诱导BMSCs,于7d用钙钴法检测碱性磷酸酶活性,10d进行茜素红矿化结节染色;在成脂诱导液中诱导BMSCs,于21d进行油红O染色。将BMSCs接种到HA/TCP植骨材料上,加入成骨诱导液,采用倒置显微镜、荧光显微镜及扫描电镜检测,并采用四氮唑蓝(MTT)比色法测定HA/TCP植骨材料上BMSCs的增殖情况。结果BMSCs在成骨诱导液中7d碱性磷酸酶呈强阳性,10d矿化结节染色呈橘红色;在成脂诱导液中,21d油红O染色呈阳性。BMSCs在HA/TCP植骨材料上孔隙周围及孔隙内生长良好并大量增殖。MTT分析结果显示,HA/TCP对BMSCs的体外增殖无抑制作用。结论BMSCs与HA/TCP植骨材料有良好的生物相容性。Objective To study the adhesion and proliferation of rabbit bone marrow mesenchymal stem cells（BMSCs） on the HA/TCP bone graft scaffold. Methods BMSCs were harvested from the iliac bone and obtained by using adhesive culture method. BMSCs were induced with osteogenie medium, at the 7th day, the cells were stained by the calcium cobalt method to show the activities of alkaline phosphatase （ALP）. At the 10th day, the mineralized nodules of osteoblasts stained with chinalizarin. Otherwise BMSCs were induced with adipogenic medium,and at the 21st day the cells stained with oil red O. We cultured the BMSCs on the surface of a matrix scaffold with osteogenic medium. The morphologic characters were checked by light microscopy, fluorescence microscopy and scanning electronic microscopy, and the proliferation of BMSCs were assayed using the MTT test. Results At the 7th day after osteoblastic induction, ALP was strongly positive, and at the 10th day, mineralized nodules stained with chinalizarin were jacinth. At the 21th day after adipogenic induction, the adipocytes were stained with oil red O. BMSCs grew well around or in pores of the HA/TCP bone graft scaffold and could be seen using both fluorescence microscopy and scanning electron microscopy. The MTT test assay showed that the HA/TCP couldn＇ t inhibit the proliferation of BMSCs. Conclusion BMSCs have a good biocompatibility with a HA/TCP bone graft scaffold.

关 键 词：骨髓间充质干细胞 羟基磷灰石 磷酸三钙 诱导 扫描电镜 兔 

分 类 号：R322.8[医药卫生—人体解剖和组织胚胎学]