国产雷帕霉素药物洗脱支架体外细胞毒性研究

目的: 比较国产雷帕霉素药物洗脱支架的体外细胞毒性的差异,为国产药物洗脱支架的细胞毒性标准的制定和体外安全性评价提供参考。 方法: 将国产雷帕霉素药物洗脱支架制备成相应的浸提液,与L929 细胞接触培养,通过倒置显微镜观察其形态,采用MTT(四唑盐) 比色法量化细胞毒性,计算相对增殖率(RGR) ,并进行细胞毒性评价。 结果: I号支架第5天浸提液的细胞相对增殖率为47.3%±2.84%,细胞毒性分级为3级;第6天浸提液的细胞相对增值率为119.2%±7.1%,细胞毒性分级为0级。II号支架第5天浸提液的细胞相对增殖率为55.5%±3.7%,细胞毒性分级为2级,第6天浸提液的细胞相对增值率为61.4%±4.7%,细胞毒性分级为2级;第7天浸提液的细胞相对增值率为117.1%±10.1%,细胞毒性分级为0级;将I号支架24 h浸提液稀释2倍至稀释256倍后其细胞毒性均为2级,稀释512倍后,其细胞毒性为1级;将II号支架24 h浸提液稀释4倍至稀释64倍后其细胞毒性均为2级,稀释128倍至512倍,其细胞毒性为1级或0级。 结论: 当国产雷帕霉素药物洗脱支架的药物释放到第6~7天或者将所含药物浓度稀释一定倍数后,其细胞毒性明显减少,表明其细胞毒性呈剂量依赖性。

Objective: To compare the cellular toxic response with respect to the different sirolimus-eluting stents and evaluate the biocompatibility of them. Methods :Two sirolimus-eluting stents were incubated into serum-supplemented minimum essential medium for different days and different doses.L929 cells were used.The cellular morphology was observed by phase contrast inverted microscopy and proliferation of the cells was examined using mitochondrial function (MTT) assay.Relative growth rate (RGR) was calculated and the biological reactivity was described and rated on a scale of 0 to 4. Results: The observed response to NO.I sirolimus-eluting stent for the 5th day was grade 3 and for the 6th day was grade 0.The observed responses to NO.II sirolimus-eluting stent for the 5th day and the 6th day were both grade 2 and for the 7th day was grade 0.The biological reactivity was grade 1 when the extraction fluid of NO.I sirolimus-eluting stent was diluted 512 times.The biological reactivity was grade 1 when the extraction fluid of NO.II sirolimus-eluting stent was diluted 128 times. Conclusions: It showed the responses to sirolimus-eluting stent were grade 0 or 1 when sirolimus were eluted or diluted.