不同孔径β-TCP对大鼠骨髓间充质干细胞增殖及成骨作用的体外研究  

Effect of various pore sizes of porous beta-tricalcium phosphate on proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cell: An in vitro study

作　　者：刘楠[1] 李海[1] 赵黎[1] 胡玲珑[1] 

机构地区：[1]上海交通大学医学院附属新华医院小儿骨科,200092

出　　处：《国际骨科学杂志》2012年第6期413-416,共4页International Journal of Orthopaedics

基　　金：教育部高等学校博士学科点专项科研基金(20090073120037);上海市科委医学引导类项目(09411962600);上海市卫生局青年科研项目(2008Y069)

摘　　要：目的探讨不同孔径β-磷酸三钙(β-TCP)对大鼠骨髓间充质干细胞(BMSC)增殖及成骨分化的作用。方法体外原代培养BMSC,取第2代细胞悬液100μl(细胞浓度2×106/ml)接种于5组不同孔径(187～300、300～375、375～500、500～750、750～830μm)的圆盘状多孔β-TCP材料中。采用细胞计数试剂盒(CCK-8)测定BMSC接种后3、24、48、72h增殖情况;扫描电镜观察接种72h后BMSC在多孔β-TCP上贴附、生长情况;碱性磷酸酶(ALP)试剂盒检测接种72h后BMSC的ALP活性。结果接种后48hBMCS在各组多孔β-TCP上均有显著增殖,其中500～750μm组增殖最快(P<0.05);扫描电镜观察显示,BMCS在多孔β-TCP材料上黏附、伸展、聚集,生长状态良好;接种后72h各组BMCS的ALP活性均明显增高,其中500～750μm组ALP活性最高(P<0.05)。结论不同孔径的多孔β-TCP对BMSC增殖及成骨分化均有促进作用,其中500～750μm孔径的促进作用最明显,可能是促进BMSC增殖及成骨分化的最适孔径。Objective To investigate the effect of various pore sizes of porous beta-triealcium phosphate （β-TCP） on the proliferation and osteogenie differentiation of the bone marrow mesenchymal stern cells （BMSCs） in rat. Methods BMSCs were isolated from rat femur and cultured in vitro. The same volume of 100μl of second passage cells suspension with concentration of 2 × 10^6/ml were implanted in porous β-TCP with disc shape and various pore sizes （18-230μm, 300-375 μm, 375-500μm, 500- 750μm, 750 830μm）. Cell proliferation was evaluated at 3, 24, 48, 72 hours after implantation by cell counting kit-8 （CCK-8）. The morphology of BMSCs growing on β-TCP was observed by scanning electron microscope （SEM）. Osteogenic differentiation of BMSCs was evaluated in terms of activity of alkaline phosphatase （ALP） in cells at 72 hours after implantation by ALP assay. Results BMSCs in all groups grew fast at 48 hours after implantation, while the cells in group of 500-750μm showed the highest proliferation （P〈0. 05）. Good adhesion, extension and aggregation of the cells were also demonstrated in each group by means of SEM. ALP activity of BMSCs in all groups increased at 72 hours after implantation, while cells in group of 500-750μm increased most significantly （P〈0. 05）. Conclusions Various pore sizes （pore sizes range： 187-830μm） of porous β-TCP could enhance the proliferation and osteogenic differentiation of BMSCs implanted, the highest improvement has been found in the porous β-TCP with pore size of 500-750μm, which is supposed to be the optimal pore size for growth and osteogenie differentiation of BMSCs.

关 键 词：Β-磷酸三钙 骨髓间充质干细胞 细胞增殖 碱性磷酸酶活性 体外研究 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]