珊瑚羟基磷灰石材料对富血小板血浆诱导培养人骨髓基质干细胞成骨特性的影响

Effect of coral hydroxyapatite on the osteogenic property of platelet-rich plasma-induced human bone marrow stromal stem cells

背景：前期实验中发现富血小板血浆能有效促进人骨髓基质干细胞的增殖及诱导分化，且对细胞的生物学特性无明显影响。目的：拟进一步验证富血小板血浆诱导培养的人骨髓基质干细胞与珊瑚羟基磷灰石材料复合后对细胞生长的影响。设计、时间及地点：单一样本观察，于2007-10／2008—04在中山大学实验室完成。材料：YHJ500珊瑚羟基磷灰石材料：多孔，孔隙相通，孔径范围280～370μm，平均325μm。方法：抽取6名健康志愿者自体外周静脉血获取富血小板血浆。采用密度梯度离心法获取健康志愿者骨髓间充质干细胞。将由富血小板血浆传代培养的人骨髓基质干细胞按照5×10^6的浓度与YHJ500珊瑚羟基磷灰石材料复合。主要观察指标：分别于第2，4，6，8天检测细胞形态学、与材料复合后细胞增殖、细胞微量蛋白及碱性磷酸酶含量。结果：与材料复合培养的人骨髓基质干细胞复合后，12-24h可见到细胞开始与材料黏附，随时间延长材料表面黏附细胞增多，材料周边细胞增殖速度及形态未受到明显影响。细胞微量蛋白含量与碱性磷酸酶测定显示单纯细胞培养组与细胞材料复合培养组人骨髓基质干细胞的细胞微量蛋白水平无明显差异，且均随时间的延长而增加。结论：富血小板血浆诱导培养的人胃髓基质干细胞生物学特性无明显变化，其与YHJ500珊瑚羟基磷灰石材料生物相容性好，YHJ500珊瑚羟基磷灰石材料适合作为组织工程的支架材料。

BACKGROUND： Studies have reported that, platelet-rich plasma is effective to promote the proliferation and differentiation of human bone marrow stromal stem cells （BMSCs）. Moreover, platelet-rich plasma has no impact on the biological property of BMSCs. OBJECTIVE： To verity the growth of BMSCs induced by platelet-rich plasma by the combination with coral hydroxyapatite. DESIGN, TIME AND SETTING： Single sample observation was performed in the laboratory of Sun Yat-sen University from October 2007 to April 2008. MATERIALS： YHJ500 coral hydroxyapatite was porous and the pores were interconnected, pore size ranged 280-370 μm with an average of 325 μm. METHODS： Platelet-rich plasma was harvested from peripheral vein in six health volunteers. BMSCs were obtained using density gradient centrifugation from health volunteers. Subsequent to the passage culture, the BMSCs were combined with YHJS00 coral hydroxyapatite at a density of 5×10^6. MAIN OUTCOME MEASURES： At 2, 4, 6 and 8 days, the cellular morphology, cellular proliferation, cell microprotein level and alkaline phosphatase were determined. RESULTS： After the BMSCs were compound cultured with the materials for 12-24 hours, cells began to adhere to materials and the adherent cells increased along with the time. The proliferation speed and morphology of the cells surrounding the materials had not been influenced. Cell microprotein level and alkaline phosphatase were determined and there was no significant difference in the microprotein level of the BMSC between culture control group and compound culture group, the microprotein level increased with the time. CONCLUSION： The human BMSCs induced by platelet-rich plasma has no changes of biological property, and is well biocompatible to YHJ500 coral hydroxyapatite. YHJ500 coral hydroxyapatite is suitable as a scaffold material of tissue engineering.