磷酸钙/聚乳酸羟基乙酸复合骨水泥对破骨前体细胞炎症因子表达的影响

Effects of calcium phosphate / poly( lactic-co-glycolic acid) composite cement on expression of inflammation factors in osteoclast precursors

目的观察磷酸钙/聚乳酸羟基乙酸复合骨水泥(CPC/PLGA)对破骨前体细胞炎症因子TNF-α、IL-1、IL-6表达的影响,了解PLGA促进CPC降解的机理。方法 1根据高效液相色谱法检测出的乳酸、羟基乙酸浓度配置CPC/PLGA替代液。2将RAW264.7细胞分别培养在DMEM培养基(对照组),第6小时CPC初凝块浸提液(A组)及CPC/PLGA替代液(B组)。通过CCK-8法检测各组细胞的增殖变化,并通过RT-PCR法检测各组中TNF-α、IL-1、IL-6 mRNA的表达情况。结果增殖实验结果示,A组细胞的增殖率在5 d时达到最高值(1.01±0.02),而B组细胞的增殖率在5 d时达到最高值(1.07±0.02),均与对照组相比有统计学意义(P<0.05)。RT-PCR法示A组和B组细胞TNF-αmRNA、IL-1 mRNA在6 h时表达最高,IL-6mRNA在12 h时表达最高,与对照组比较均有统计学差异(P<0.05)。结论磷酸钙/聚乳酸羟基乙酸复合骨水泥调控破骨前体细胞炎症因子增高。

Objective To investigate the effects of calcium phosphate / poly（ lactic-co-glycolic acid） composite cement（ CPC / PLGA） on expression of inflammation factors TNF-α,IL-1,IL-6 in osteoclast precursors,and to understand the mechanism of PLGA in promoting the degrading of CPC. Methods 1 According to the concentration of lactic acid and glycolic acid detected by high performance liquid chromatography（ HPLC） method,CPC / PLGA alternative solution was configurated. 2 RAW264. 7 cells were cultured in DMEM medium（ control group）,6 h CPC clot soaking liquid（ group A） and CPC / PLGA alternative liquid（ group B）. The proliferation of RAW264. 7 cells were tested by CCK-8 method. Then the cells from each group were stimulated,and subsequently the mRNA expressions of TNF-α,IL-1,IL-6 were analyzed through RT-PCR. Results Proliferation assay revealed that the cell proliferation of group A reached its maximum which was 1. 01 ± 0. 02 at 5 d,while group B at 5 d was 1. 07 ± 0. 02,and compared to control group,the differences were statistically significant（ P〈0. 05）; RT-PCR showed that TNF-α mRNA,IL-1mRNA expression in 6 h was highest in group A and group B,while IL-6 mRNA expression in 12 h was highest,and compared with the control group the differences were statistically significant（ P〈0. 05）. Conclusion CPC / PLGA can increase the expression of inflammation factors in osteoclast precursors.