两种多孔生物玻璃支架材料的体外细胞相容性研究

Cytocompatibility of two porous bioactive glass-ceramic in vitro

目的对比研究采用同样原料但以不同方法制备的两种多孔生物玻璃支架材料的体外细胞相容性。方法溶胶-凝胶法制备A/W生物活性微晶玻璃(4006材料),熔融法制备多孔生物活性玻璃(45S5材料)。体外诱导分离培养及鉴定兔骨髓基质细胞(BMSCs),通过材料浸提液细胞毒性实验(MTT法)、细胞黏附实验、倒置相差显微镜、扫描电镜和环境扫描电镜比较4006材料和45S5材料对BMSCs细胞黏附和生长的影响,并探索与材料复合的最佳BMSCs细胞悬液浓度。结果 4006材料浸提液培养1 d时,其细胞活力与纯培养液间无统计学差异(P>0.05);但培养3 d后,其细胞活力低于纯培养液(P<0.01)。45S5材料浸提液培养的细胞活力明显低于纯培养液(P<0.01)。细胞与材料复合培养后,镜下见BMSCs在4006材料孔隙内贴壁生长良好,分泌基质活跃;而在45S5材料上细胞黏附生长较差。BMSCs与4006材料的黏附量随接种细胞浓度升高而升高,细胞悬液浓度为2×107个.mL-1时的细胞黏附量最高。结论溶胶-凝胶法制备的A/W生物活性微晶玻璃具有良好的生物活性和细胞相容性,具有作为骨组织工程支架材料的潜能。与其复合的细胞悬液浓度需要2×107个.mL-1或以上。

Objective To compare the cytocompatibility of two kinds porous bioactive glass-ceramic made by same raw materials. Methods Apatite/wollastonite bioactive glass-ceramic （4006） were prepared by sol-gel method, and bioactive glass （45S5） were prepared by melting method. Bone marrow stromal cells （BMSCs） were cultivated, differen- tiated and proliferated into osteoblasts, from a rabbit＇s marrow in the differentiation culture medium with active func- tion. The viability of BMSCs cultivated with extraction of these two kinds of biomaterial, which could represent the cytotoxicity effect of 4006 and 45S5 against BMSCs, was evaluated by the MTY assay. BMSCs were seeded and co- cultivated with two kinds of biomaterial scaffolds respectively in vitro. The proliferation and biological properties of cells adhered to scaffolds were observed by inverted phase contrast microscope, scanning electron microscope （SEM）, and environmental scanning electron microscope （ESEM）, and a suitable cell amount for seeding on the scaffold was searched. Results There was no difference on the viability of BMSCs only cultured for one day by complete extract of 4006 and culture medium （P〉0.05）, but there was significant difference between them when the ceils had been cultured for 3 days （P〈0.01）. The extract of 45S5 had significantly higher cytotoxicity than extract of culture medium （P〈0.01）. The BMSCs adhered, spread, and proliferated throughout the pores of the scaffold 4006, and the amountof cells adhered to 4006 was more than to 45S5. The adhered cells to 4006 increased with the rising amount of cells seeded. And 2×10^7 cells·mL^-1 suspension resulted in the highest eel1 adherence during the comparative cell adherence test. Conclusion Apatite/wollastonite bioac-tive glass-ceramic has good bioactivity and cytocompatibility. Therefore, it may have the potential to be a new cell vehicle for bone tissue engineering. And the suitable seeding cell amount of apatite/wollastonite bioactive glass-ceramic should be 2×l0^7cells·mL^-1 or even more than that.