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

摘要

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-6 mRNA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/PL-GA) 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 per-formance liquid chromatography ( HPLC) method, CPC/PLGA alternative solution was configurated. ② RAW264. 7 cells were cul-tured 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 prolifera-tion 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-αmRNA, 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.