恒河猴骨髓基质干细胞与新型可吸收羟基磷灰石及β-磷酸三钙体外相容性的观察

In vitro biocompatibility of novel absorbable hydroxyapatite and AO artificial bone β-tricalcium phosphate with rhesus bone marrow stromal cells

目的观察恒河猴骨髓基质干细胞(rBMSC)与新型可吸收羟基磷灰石(HA)及AO人工骨茁-磷酸三钙(茁-TCP)的体外相容性。为在灵长类动物体内构建组织工程化骨作材料方面的准备。方法取第三代恒河猴骨髓基质细胞与材料复合培养。实验分3组:A组将rBMSC与HA复合培养;B组rBMSC与茁-TCP复合培养;C组为对照,只有细胞不加材料。倒置相差显微镜、扫描电镜观察各组细胞形态及增殖情况,MTT法半定量检测细胞增殖。结果倒置显微镜下见HA组细胞生长良好,与对照组无显著差异。茁-TCP组有一些细小颗粒脱落散在分布于板底或细胞表面,有少量细胞脱落死亡。扫描电镜见HA组细胞贴附、增殖良好,茁-TCP组细胞贴附率不高。MTT法显示HA组细胞增殖与对照组接近,而茁-TCP组则显著低于对照组。结论新型HA与恒河猴BMSc生物相容性好,可用作恒河猴骨组织工程的支架材料,AO人工骨需要作性能上的改进。

Objective To study the in vitro biocompatibility of novel hydroxyapatite (HA) and AO artificial bone β-tricalcium phosphate (β-TCP) with rhesus bone marrow stromal cells (rBMSCs) . Methods The third passage of rBMSCs were cultured with HA and β-TCP respectively, with the cells cultured without the materials as the control. The morphology and proliferation of cells were observed by inverted phase-contrast microscope and scanning electron microscope (SEM). MTT assay was used to semiquantitatively evaluate the cell proliferation. Results The rBMSC cocultured with HA exhibited good growth as observed under inverted phase-contrast microscope, without significant difference from the cells in the control group. Some small particles were seen pealing off from β-TCP, and some of the cells died. Under SEM, rBMSCs showed good adhesion to HA with obvious proliferation, but the ratio of adhesive cells was not as high as that in β-TCP group. MTT assay showed no significant difference in the cell number between HA and the control groups, but the cell number in β-TCP group was notably less than that of control group. Conclusion Novel HA has good biocompatibility with rBMSCs for bone tissue engineering, and AO artificial bone still needs improvement to serve as scaffold material for BMSCs.