新型有序纳米介孔生物玻璃材料对体外培养成骨细胞的影响

Influence of mesoporous bioactive glass on proliferation, adhesion, differentiation and mineralization in osteoblasts in vitro

目的探讨新型有序纳米介孔生物活性玻璃材料(mesoporous bioactive glass,MBG)对体外培养成骨细胞(osteoblast,OB)粘附、增殖、分化及矿化的影响。方法用混合酶消化法分离大鼠颅盖骨成骨细胞,进行原代培养;成骨细胞与不同浓度的MBG浸提液作用后,MTT法测定细胞增殖;PNPP法检测碱性磷酸酶活性;矿化结节计数和面积测量分析MBG对成骨细胞矿化能力的影响。同时扫描电镜对直接在MBG材料上培养的OB进行细胞形态学和粘附情况分析。结果0.5%以下各浓度组MBG浸提液均对成骨细胞增殖无影响,而1%浓度组对细胞增殖有轻度抑制,与对照组相比差异有统计学意义(P<0.01);各浓度MBG浸提液均可以增强碱性磷酸酶活性,与对照组相比,其中0.0625、0.125、0.25%浓度明显提高ALP活性,差异有统计学意义(P<0.01);0.0625%浓度时,矿化结节数目差异无统计学意义(P>0.05),而矿化结节面积差异有统计学意义(P<0.01);扫描电镜下细胞形态分化良好,细胞相互之间及细胞与材料之间结合良好。结论不同浓度MBG浸提液对成骨细胞的增殖和分化有不同的作用,低浓度对成骨细胞的增殖和分化有更好的作用。MBG材料有良好的生物相容性,有望成为新型骨组织修复替代材料或骨组织工程支架材料。

Objective To investigate the effects of mesoporous bioaetive glass （MBC） on proliferation, adhesion, differentiation and mineralization in osteuhlasts in vitro. Methods Osteoblastie cells were harvested by sequential enzyme digestion from calvaria of Sprague Dawley rats and then cultured. They were treated with leaching liquor of mesoporous bioaetive glass to analyze proliferation MTT methods. PNPP method was used to assay ALP activity and analyze the mineralization ability of nsteoblasts. The number and area of mineralizatinn nodes were ohserved. Other nsteoblasts were cultured on the MBG plate to evaluate cellular morphology, and adhesion by SEM. Results MBG did not affect the proliferation of osteoblasts except at the concentration of 1% （P〈0.01）. The ALP activity was higher at all concentrations, especially at 0.0625, 0.125 and 0.25% , as compared to control （P〈0.01）. MBG significantly enhanced osteoblastic mineralization at the concentration of 0.0625%. Viable osteoblasts were seen on MBG, presenting with perfect adhesion. Conclusions Different concentrations of MBG presented different effects on proliferation, differentiation and mineralization in osteoblasts in vitro. MBG showed perfect hiocompatibility, suggesting thai MBG is a prnmisiag material for hone repair and tissue engineering.