目的:研究体外雷奈酸锶对钛(Ti)颗粒刺激单核/巨噬细胞分泌溶骨因子及其膜上RANK表达的影响,探讨雷奈酸锶预防人工关节假体周围无菌性松动的可能性。方法:体外培养单核巨噬细胞白血病细胞RAW264.7,制备Ti颗粒,并采用MTT法检测绘制RAW264.7细胞增殖曲线,寻找雷奈酸锶最佳干预浓度。将RAW264.7分为3组:Ti微粒组(体积分数为0.1%的含Ti微粒+质量分数为10%的胎牛血清DMEM培养基)、Ti+雷奈酸锶组(体积分数为0.1%的含Ti微粒+最佳浓度雷奈酸锶+含质量分数为10%的胎牛血清DMEM培养基)和对照组(含质量分数为10%的胎牛血清DMEM常规培养基)。采用ELISA法检测各组培养液白细胞介素-1(IL-1)、肿瘤坏死因子-α(TNF-α)浓度。采用实时荧光定量SYBR GREEN法检测RANK m RNA表达。结果:Ti微粒组及Ti+雷奈酸锶组的细胞培养液中IL-1、TNF-α的含量及RAW264.7细胞RANK m RNA的表达量明显高于对照组(P〈0.05)。而Ti+雷奈酸锶组的IL-1、TNF-α的含量及RAW264.7细胞RANK m RNA的表达量明显低于Ti微粒组(P〈0.05)。结论:Ti颗粒能刺激单核/巨噬细胞分泌IL-1、TNF-α,并能促进单核/巨噬细胞表达RANK。雷奈酸锶能明显抑制Ti颗粒诱导单核细胞分泌IL-1、TNF-α,以及抑制单核/巨噬细胞表达RANK。
[ABSTRACT]Objective:To study the effects of strontium ranelate in vitro on titanium(Ti)particles stimulating mononuclear macrophage to secrete osteolysis factor and its RANK expression in order to probe the possibility of strontium ranelate to prevent periprosthetic aseptic loosening.Methods:Cultured in vitro mononucle-ar macrophage leukemia cells RAW264.7,prepared Ti particles,and detected and drew the cell proliferation curve of RAW264.7 with MTT method,to look for the best intervention concentration of strontium ranelate.RAW264.7 were divided into 3 groups:a Ti group(Ti particles with the volume ratio of 0.1% + 10% FBS DMEM),a Ti + strontium ranelate group(Ti particles with the volume ratio of 0.1% + strontium ranelate with best concentra-tion + 10% FBS DMEM)and a control group(10% FBS DMEM).The ELISA method was used to detect the concentration of interleukin-1(IL-1)and tumor necrosis factor-α(TNF-α)in each culture solution.Detected the expression of RANK mRNA with the real-time lfuorescence quantitative SYBR GREEN method.Results:The content of IL-1 and TNF-α and the expression of RANK mRNA of RAW264.7 cells in the cell culture fluid of the Ti group and the Ti + strontium ranelate group were signiifcantly higher than those in the control group (P 〈 0.05).While the content of IL-1 and TNF-α and the expression of RANK mRNA of RAW264.7 cells in the cell culture lfuid of the Ti + strontium ranel-ate group were signiifcantly lower than those in the Ti group,the difference between thembeing statistically significant(P 〈 0.05).Conclusion:Ti particles can stimulate mononuclear macrophage to secrete IL-1 and TNF-α and its RANK expression.Strontium ranelate can obviously inhibit Ti particles to secrete IL-1,TNF-α and the expression of RANK.