人牙髓细胞复合不同孔径羟基磷灰石/磷酸三钙支架材料的生物学行为  

Biological behavior of human dental pulp cells on hydroxyapatite/tricalcium phosphate scaffold with different pore sizes

作　　者：彭伟伟[1] 翟万银[2] 江龙[1] 李莉芬[1] 常江[2] 朱亚琴[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院口腔综合科上海市口腔医学重点实验室,上海市200011 [2]中国科学院上海硅酸盐研究所生物材料与组织工程中心,上海市200050

出　　处：《中国组织工程研究与临床康复》2010年第51期9567-9571,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家教育部出国留学人员基金(2008-0890-09)"人牙髓细胞CXCR4+细胞的筛选;鉴定和诱导分化";上海市科学技术委员会项目(08DZ2271100"上海市口腔医学重点实验室";08JC1414500"SDF-1/CXCR4轴在修复性牙本质形成中的作用机理研究");上海市教委科研创新重点项目(09ZZ116)"人牙髓CXCR4+细胞分选;鉴定和增殖分化研究"~~

摘　　要：背景:有文献表明,通过组织工程的方法将人牙髓细胞复合羟基磷灰石/磷酸三钙多孔支架材料修复牙缺损具有可行性。然而究竟多大孔径的支架材料最有利于人牙髓细胞的生长及分化,至今尚无定论。目的:观察人牙髓细胞复合不同孔径羟基磷灰石/磷酸三钙支架材料后黏附、增殖和分化等生物学行为。方法:人牙髓细胞接种至3种不同孔径的羟基磷灰石/磷酸三钙材料上,采用荧光显微镜以及扫描电镜检测细胞在材料表面的黏附生长情况,然后通过细胞的黏附率实验与MTT比色法观察人牙髓细胞在材料表面的黏附与增殖特性。不同孔径的羟基磷灰石/磷酸三钙支架复合人牙髓细胞后分别用生长培养基和矿化诱导液培养,于接种后第4,7,10天检测碱性磷酸酶活性。结果与结论:人牙髓细胞在3种不同孔径支架材料表面和孔隙内均能顺利黏附并增殖。其中100～300μm组支架材料黏附率最高,MTT结果显示接种3d后300～500μm组能较好地促进细胞增殖。培养10d后,复合在100～300μm和300～500μm材料上的人牙髓细胞,其碱性磷酸酶活性显著高于500～700μm组。提示与500～700μm孔径相比,孔径为100～300μm和300～500μm的羟基磷灰石/磷酸三钙材料能更好地促进人牙髓细胞黏附、增殖和分化。BACKGROUND：Studies prove the feasibility of repairing dental defects using human dental pulp cells combined with hydroxyapatite/tricalcium phosphate （HA/TCP） scaffolds by means of tissue engineering methods.However,it is not clear that which pore size of scaffold can promote the growth and differentiation of dental pulp cells well.OBJECTIVE：To study the biological behaviors,such as adherence,proliferation and differentiation of human dental pulp cells on the HA/TCP scaffold with different pore sizes.METHODS：Human dental pulp cells were seeded into the HA/TCP scaffold with three kinds of pore sizes.Fluorescence microscope and scanning electronic microscope were used to observe the adherence and growth of cells to HA/TCP.Subsequently,the adherence and proliferation were assayed by adhesion rate and MTT tests,respectively.The complex of human dental pulp cells and HA/TCP scaffolds with different pore sizes were cultured with both growth medium and osteogenic medium,then the activity of alkaline phosphatase was determined at 4,7 and 10 days.RESULTS AND CONCLUSION：Human dental pulp cells adhered to the surface and the pores of HA/TCP scaffold successfully,and proliferated well.The highest rate of adhesion was observed in the group of the scaffold with pore size in the range of 100-300μm.Results of MTT showed that 300-500μm pore size group could promote cellular proliferation after 3 days.The alkaline phosphatase activity of human dental pulp cells,compound with the scaffolds with pore size of 100-300μm and 300-500μm,was significantly higher than 500-700μm group after 10 days incubation.The scaffold with the pore size in range of either 100-300μm or 300-500μm promoted the adhesion,proliferation and differentiation of human dental pulp cells better than the group of 500-700μm.

关 键 词：人牙髓细胞 羟基磷灰石/磷酸三钙 孔径 黏附 增殖 分化 

分 类 号：R318[医药卫生—生物医学工程]