新型铁基可降解生物材料的体外细胞毒性评价研究  

Study on in vitro cytotoxicity of novel iron-based biodegradable biomaterials

作　　者：盖潇潇 朱艺馨 孙晓霞 屈秋锦 张贵 刘成虎 GAI Xiao-xiao;ZHU Yi-xin;SUN Xiao-xia;QU Qiu-jin;ZHANG Gui;LIU Cheng-hu(Shandong Institute of Medical Device and Pharmaceutical Packing Inspection,NMPA Key Laboratory for Safety Evaluation of Biomaterials and Medical Devices,Shandong Key Laboratory of Biological Evaluation for Medical Devices,Jinan 250101,China;Biotyx Medical(Shenzhen)Co.,Ltd.,Shenzhen 518109,Guangdong Province,China)

机构地区：[1]山东省医疗器械和药品包装检验研究院,国家药品监督管理局生物材料器械安全性评价重点实验室,山东省医疗器械生物学评价重点实验室,济南250101 [2]元心科技(深圳)有限公司,广东深圳518109

出　　处：《医疗卫生装备》2025年第3期35-41,共7页Chinese Medical Equipment Journal

基　　金：国家重点研发计划课题(2016YFC1103205)。

摘　　要：目的:针对新型铁基可降解生物材料的降解特性,探究科学且适用于铁基可降解生物材料体外细胞毒性的评价方法。方法:选取含铁量高于99%、含氮量为0.03%~0.20%的支架半成品渗氮铁管和纯铁管作为试验样品,并将其按照不同的操作分为厌氧处理组、常规处理组、常规处理除铁颗粒组、常规处理洗板组。其中,厌氧处理组在厌氧工作站中进行厌氧处理;常规处理组在生物安全柜中进行常规处理;常规处理除铁颗粒组进行常规处理后再经Midimacs Starting Kit手动分选器进行除铁处理;常规处理洗板组先用0.9%氯化钠注射液进行洗板处理,再加入四唑盐试剂进行常规处理。采用不同浸提介质(模拟体液、细胞培养液、磷酸盐缓冲液、0.9%氯化钠注射液)对各组试验样品进行浸提处理,比较不同组中铁基可降解生物材料的降解产物对L929细胞存活率的影响。结果:厌氧处理组和常规除铁颗粒组无潜在的细胞毒性。常规处理组台盼蓝染色显示有明显的细胞毒性,但四唑盐试剂与降解产物反应导致出现假阴性。常规处理洗板组可以消除铁颗粒带来的假阴性,但仍表现出潜在的细胞毒性,而且会洗掉部分细胞,影响结果的准确性。结论:该研究提出的方法适用于新型铁基可降解生物材料的体外细胞毒性评价,可为该类材料的应用安全性评价提供重要支撑。Objective To propose an in vitro cytotoxicity evaluation method for novel iron-based biodegradable mateirals focusing on their degrading characteristics.Methods Half-finished scaffolds of nitrided iron tubes with a higher iron content of over 99%and a nitrogen content of 0.03% to 0.20% were selected as test samples,and based on the treatment mode were divided into an anaerobic treatment group,a non-anaerobic treatment group,a non-anaerobic treatment with removed iron particle group and a non-anaerobic treatment with plate-washing group.The anaerobic treatment group was processed in an anaerobic workstation;the conventional treatment group underwent standard handling in a biosafety cabinet;the conventional treatment with removed iron particle group was subjected to iron particle elimination using a Midimacs Starting Kit manual sorter after conventional treatment;and the conventional treatment with plate-washing group was rinsed with 0.9% sodium chloride injection before tetrazolium salt reagent treatment.Different extraction media(simulated body fluid,cell culture medium,phosphate buffer and 0.9% sodium chloride injection)were used for the immersion treatment of the test samples in each group to compare the effects of the degradation products on the survival rate of L929 cells.Results The anaerobic treatment group and the conventional treatment with removed iron particle group exhibited no detectable cytotoxicity.Trypan blue staining revealed significant cytotoxicity in the conventional treatment group,while false-negative results emerged due to interactions between the tetrazolium salt reagent and degradation products.In the non-anaerobic treatment with washing plate group,the false negative from iron particles was eliminated while potential cytotoxicity was showen,and the result accuracy was affected because some cells were washed out.Conclusion The proposed method can be used for the in vitro cytotoxicity evaluation,and facilitates the safety evaluation of the application of such materials.

关 键 词：铁基 可降解生物材料 体外细胞毒性 可降解金属支架 渗氮铁管 纯铁管 

分 类 号：R318.08[医药卫生—生物医学工程]