不同骨水泥材料在炎症条件下对小鼠成纤维细胞L929的生物安全性研究

Research on biological biosafety of different bone cement materials to mouse fibroblast L929 under the inflammatory conditions

目的评价不同骨水泥材料在炎症条件下对小鼠成纤维细胞L929的生物安全性。方法分别采用CCK-8法、DAPI荧光染色法、FITC/PI凋亡染色法检测3种骨水泥材料对炎症条件下小鼠成纤维细胞L929细胞活性、细胞分裂生长及细胞凋亡的影响,同时利用RT-PCR和Western blot分别检测3种骨水泥材料对炎症条件下小鼠成纤维细胞L929中炎症相关因子IL-1、IL-2、IL-6、TNF-α、IFN-γ的mRNA水平和蛋白表达水平。结果聚甲基丙烯酸甲酯(PMMA)、纳米羟基磷灰石/聚酰胺66(n-HA/PA66)、磷酸钙骨水泥(CPC)在炎症条件下均能够显著抑制小鼠成纤维细胞L929的活性和增殖,CPC抑制最显著(P<0.05);PMMA、n-HA/PA66、CPC在炎症条件下均能够显著促进小鼠成纤维细胞L929的凋亡(P<0.05),3种骨水泥材料之间差异无统计学意义(P>0.05);3种骨水泥材料在炎症条件下对小鼠成纤维细胞L929不同炎症因子表达的影响不同,但CPC相对于PMMA、n-HA/PA66更能刺激细胞多种炎症因子的表达(P<0.05)。结论不同骨水泥材料在炎症条件下对小鼠成纤维细胞L929的生物安全性不同,其中PMMA、n-HA/PA66差别不大,而CPC在炎症条件下对小鼠成纤维细胞L929的影响最显著。

Objective To evaluate biosafety of different bone cement materials to mouse fibroblast L929 under the in- flammatory conditions. Methods The cell activity, the influence of the growth of cell division and cell apoptosis of mouse fibro- blast L929 treated by different bone cement materials under the inflammatory conditions were detected by the methods of CCK 8, DAPI staining fluorescence staining, FITC/PI apoptosis respectively. Expression of IL-1, IL-2, IL-6, TNF-a and IFN-7 of mouse fibroblast L929 treated by different bone cement materials under the inflammatory conditions were analyzed by RT-PCR and Western blot. Results The activity and proliferation of mouse fibroblast L929 were significantly inhibited by PMMA, n- HA/PA66 and CPC under the inflammatory conditions （P^0.05）, and CPC were the most; The apoptosis of mouse fibroblast L929 were significantly inhibited by PMMA, n-HA/PA66 and CPC （P〈0.05）, and there were no significant differences between three types of bone cement material （P〉0.05） ; The influence of different inflammatory factor expression in mouse fibro- blast L929 were different by the three types of bone cement material under the condition of inflammation. Relative to PMMA and n-HA/PA66, the CPC stimulates the expression of different inflammatory factor in cell more （P^O. 05）. Conclusion Dif- ferent bone cement material have different biological biosafety on mouse fibroblast L929 under the condition of inflammation, there are no significant differences between PMMA and n-HA/PA66, and CPC has most significant influence on mouse fibroblast L929 under the condition of inflammation.