不同浸提条件对医疗器械体外细胞毒性试验结果的影响
The Effect of Different Extraction Conditions on the in Vitro Cytotoxicity Test Results of Medical Devices
王蕊 1宋伟 1贺学英 1徐爽1
作者信息
- 1. 北京市医疗器械检验研究院(北京市医用生物防护装备检验研究中心) (北京 101111)
- 折叠
摘要
目的 探讨不同浸提条件(浸提 24、72 h)对医疗器械体外细胞毒性试验中细胞生长与细胞毒性结果的影响.方法 采用含血清MEM细胞培养液作为浸提介质,按照GB/T 16886.12-2017 推荐的浸提比例,将空白对照品、阴性对照品及 5 种不同医疗器械常用材料样品在(37±1)℃条件下分别浸提 24、72 h,依据GB/T 16886.5-2017 中的MTT法进行试验.比较浸提 24、72 h对细胞生长与细胞毒性结果的影响.结果 与浸提 24 h比较,浸提72 h后空白对照组吸光度偏低,相对偏差为-3.0%,差异无统计学意义(P>0.05);被检样品试验结果显示,样品1、样品2有细胞毒性,样品3和样品5无细胞毒性,浸提24、72 h结果一致,相对增殖率(RGR)变化小;样品 4 有细胞毒性,但浸提 72 h后RGR明显降低,降低幅度为 93.7%,提示有毒物质溶出量增多.结论 结合显微镜观察细胞形态,确认延长浸提时间至 72 h对细胞生长的影响可以忽略.对于被检样品组,延长浸提时间至 72 h各样品RGR呈降低趋势.对于RGR>80%、<60%的医疗器械样品,浸提24、72 h的评价结果一致,评价风险较小.而对于RGR处于临界值(60%～80%)的医疗器械样品,浸提 24、72 h的结果可能不一致,评价风险较大,尤其是长期接触人体的医疗器械,建议采取浸提 72 h的条件进行体外细胞毒性试验,以降低评价风险.
Abstract
Objective To explore the effects of different extraction conditions(extraction for 24 and 72 hours)on cell growth and cytotoxicity results in in vitro cytotoxicity tests of medical devices.Methods With the use of MEM cell culture medium containing serum as the extraction medium,according to the recommended extraction ratio in GB/T 16886.12-2017,Blank control samples,negative control samples,and 5 different commonly used medical device material samples were extracted at(37±1)℃for 24 and 72 hours,respectively,and MTT tests were conducted according to GB/T 16886.5-2017.The effects of extraction with 24 and 72 hours of on cell growth and cytotoxicity results were compared.Results Compared with 24 hours of extraction,the absorbance of the blank control group was lower after 72 hours of extraction,with a relative deviation of-3.0%,and the difference was not statistically significant(P>0.05);The results showed that sample 1 and sample 2 showed cytotoxicity,while sample 3 and sample 5 showed no cytotoxicity.The results were consistent for 24 and 72 h after extraction,and the relative proliferation rate(RGR)changed little;Sample 4 had cytotoxicity,but the RGR decreased significantly after 72 h of extraction,with a reduction of 93.7%,indicating that the dissolved amount of toxic substances increased.Conclusions By observing the cell morphology under a microscope,it was confirmed that extending the extraction time to 72 hours had negligible effects on cell growth.For the tested sample group,the RGR of each sample showed a decreasing trend by extending the extraction time to 72 hours.For medical device samples with RGR>80%and<60%,the evaluation results after 24 and 72 hours of extraction were consistent,and the evaluation risk was relatively low.For medical device samples with RGR at the critical value(60%to 80%),the results of extraction for 24 and 72 hours might be inconsistent,and the evaluation risk was high.Especially for medical devices that are in long-term contact with human body,it is recommended to use the condition of 72 h extraction for in vitro cytotoxicity test to reduce the risk of evaluation.
关键词
细胞毒性试验/体外/MTT法/浸提Key words
Cytotoxicity tests/In vitro/MTT method/Extraction引用本文复制引用
出版年
2024
NETL
NSTL
万方数据