采用体内碱性彗星试验检测两种聚醚醚酮材料的遗传毒性

doi:10.3969/j.issn.1004-616x.2022.05.010

摘要/Abstract

摘要： 目的：采用哺乳动物体内碱性彗星试验，检测两种聚醚醚酮材料的遗传毒性，为医疗器械及其材料的遗传毒性体内碱性彗星试验方法的建立提供依据。方法：采用0.9%氯化钠注射液(SC)和棉籽油(CSO)两种介质制备聚醚醚酮材料的试验液，以SC和CSO作为阴性对照，甲基磺酸甲酯(MMS)作为阳性对照。选取 SD大鼠 70只，雌雄各半，大鼠连续两次(间隔 24 h)染毒，SC组和MMS阳性对照组按照 10 mL/kg的剂量由静脉途径染毒，CSO组按照 5 mL/kg的剂量由腹腔途径染毒，末次染毒后 3 h处死大鼠，称取肝脏和肾脏的质量并进行组织病理学检查。取肝脏、肾脏和外周血分别制备单细胞悬液进行碱性彗星试验，以尾部DNA百分比、尾矩和Olive尾矩为分析指标判断DNA损伤程度。结果：试验组的大鼠肝脏系数、肾脏系数分别与SC和CSO阴性对照组相比差异均无统计学意义(P >0.05)，所有大鼠的肝脏和肾脏形态结构正常，未见明显的组织病理学改变。与 SC和 CSO阴性对照组相比，MMS阳性对照组尾部DNA含量百分比、尾矩和Olive尾矩的差异有统计学意义(P<0.01)，两种聚醚醚酮材料组间的差异无统计学意义(P>0.05)。结论：应用体内碱性彗星试验的研究方法，在本研究条件下，两种聚醚醚酮材料的试验液不能诱导大鼠肝细胞DNA链断裂，未检测出遗传毒性。

关键词: 体内碱性彗星试验, 医疗器械, 甲基磺酸甲酯, 遗传毒性, 聚醚醚酮材料

Abstract: OBJECTIVE: An in vivo mammalian alkaline comet test was used to detect genotoxicity of two kinds of polyether ether ketone materials,and to develop the method for genotoxicity evaluation of medical devices.METHODS: Two polyether ether ketone materials were extracted with 0.9% sodium chloride injection (SC) and cottonseed oil (CSO) into rats,and used as the test solution.SC and CSO were prepared the same way and served as the negative controls.Methyl methanesulfonate (MMS) was used as positive control.70 SD rats (half males and half females) received two treatments at 24 h intervals.The SC and MMS groups of rats received treatments through intravenous injection at a dose of 10 mL/kg.The CSO group received treatments through intraperitoneal injection at a dose of 5 mL/kg.All rats were sacrificed at 3 h after the last treatments.Their livers and kidneys were weighed and histopathological examined.Single-cell suspensions were prepared from liver,kidney and peripheral blood for the alkaline comet assay which determined DNA damage via% tail DNA,tail moments and Olive tail moments.RESULTS: Under the conditions of this study,the liver and kidney coefficients of the test solution and the MMS positive control group showed no statistically significant differences compared with the SC and CSO negative control groups,respectively.The shape and structure of the liver and kidney of all rats were normal without obvious histopathological changes.There were statistically significant differences in the% tail DNA,tail moment and Olive tail moment in the MMS positive control group (P<0.01) and there were no statistically significant differences in the two polyether ether ketone materials groups (P>0.05) compared with the SC and CSO negative control groups.CONCLUSION: The method of in vivo alkaline comet assay for genotoxicity of medical devices was successfully tested.The two polyether ether ketone materials were considered unable to induce DNA strand breakage in the rat liver under the conditions of this study.

Key words: in vivo alkaline comet assay, medical device, methyl methanesulfonate, genotoxicity, polyether ether ketone materials

中图分类号:

Q355

孙令骁, 刘香东, 刘增祥, 王鸾鸾, 朱福余, 孙晓霞, 刘成虎. 采用体内碱性彗星试验检测两种聚醚醚酮材料的遗传毒性[J]. 癌变·畸变·突变, 2022, 34(5): 388-394,399.

SUN Lingxiao, LIU Xiangdong, LIU Zengxiang, WANG Luanluan, ZHU Fuyu, SUN Xiaoxia, LIU Chenghu. Application of an in vivo alkaline comet assay in genotoxicity of two polyether ether ketone materials[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2022, 34(5): 388-394,399.

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