外周血微核试验检测可降解生物材料补片的遗传毒性

doi:10.3969/j.issn.1004-616x.2023.05.010

癌变·畸变·突变 ›› 2023, Vol. 35 ›› Issue (5): 387-391.doi: 10.3969/j.issn.1004-616x.2023.05.010

外周血微核试验检测可降解生物材料补片的遗传毒性

王国伟¹, 孙晓霞¹, 车国喜¹, 盖潇潇¹, 屈秋锦¹, 王焱¹, 汪晓飞¹, 杜晓丹², 刘成虎¹

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, 山东济南 250101;
2. 中国食品药品检定研究院, 北京 102629

收稿日期:2023-05-23 修回日期:2023-09-05 发布日期:2023-10-13
通讯作者: 杜晓丹, 刘成虎
作者简介:王国伟,E-mail:383052249@qq.com。
基金资助:
“十三五”国家重点研发专项(2016YFC1103205)；山东省医疗器械和药品包装检验研究院项目(ZX202120)

Genotoxicity evaluation of biodegradable patches using the micronucleus test in peripheral blood of mice

WANG Guowei¹, SUN Xiaoxia¹, CHE Guoxi¹, GAI Xiaoxiao¹, QU Qiujin¹, WANG Yan¹, WANG Xiaofei¹, DU Xiaodan², LIU Chenghu¹

1. Shandong Institute of Medical Device and Pharmaceutical Packing Inspection/NMPA Key Laboratory for Safety Evaluation of Biomaterials and Medical Devices, Jinan 250101, Shandong;
2. National Institutes for Food and Drug Control, Beijing 102629, China

Received:2023-05-23 Revised:2023-09-05 Published:2023-10-13

摘要/Abstract

摘要： 目的： 用流式细胞术检测小鼠外周血中的网织红细胞微核(MN-RET)与成熟红细胞微核(MN-NCE)，探究可降解生物材料补片的潜在遗传毒性风险。方法： 实验设短周期和长周期给予受试物两种方案，小鼠分别腹腔注射介质对照液、样品试验液、环磷酰胺溶液(25、50 mg/kg)，每日给药1次，短周期连续给予受试物3 d后采集外周血，流式细胞术检测外周血中的微核；长周期连续给予受试物14 d后采集外周血并摘取脾脏，流式细胞术检测外周血中的微核并分析脾脏细胞的凋亡情况，同时用HE对脾脏染色后进行组织病理学检查。结果： 不同周期条件下样品试验组小鼠外周血中MN-RET百分率和MN-NCE百分率相较于介质对照组差异均无统计学意义(P>0.05)；长周期给予样品试验组小鼠脾脏细胞凋亡率相较于介质对照组差异不显著(P>0.05)，脾脏组织病理学检查无明显病理性改变。而长周期给予受试物后，与介质对照组相比，25和50 mg/kg环磷酰胺组晚期凋亡细胞率均显著升高(P<0.05或P<0.01)，且50 mg/kg环磷酰胺组小鼠脾脏出现明显的白髓萎缩、淋巴细胞减少，红髓扩张、髓外造血增加等病理性改变。结论： 该可降解生物材料补片在外周血微核试验中不存在遗传毒性风险。在给予试物不同周期条件下，选择分析小鼠外周血中的MN-RET和MN-NCE，在可降解生物材料补片的遗传毒性风险研究中有重要的意义。

关键词: 流式细胞术, 微核试验, 遗传毒性, 可降解生物补片

Abstract: OBJECTIVE:Based on flow cytometry,genotoxicity of biodegradable patch was investigated by studying the induction of micronuclei in peripheral reticulocytes (MN-RET) and mature erythrocytes (MN-NCE) in mice under different administration conditions. METHODS: For the short-term administration condition, mice were intraperitoneally injected with vehicle control,test solution,and cyclophosphamide solution (25,50 mg/kg) once a day for 3 days. Then,peripheral blood samples were collected,and the micronucleus were detected by flow cytometry. For the long-term administration condition, mice were continuous administrated for 14 days. Then,peripheral blood samples were collected,and spleens were enucleated. Micronucleus frequencies as well as splenic cell apoptosis were detected by flow cytometry. In addition, spleens were histopathologically examined after hematoxylin-eosin staining (HE). RESULTS: There was no significant differences in the percentage of MN-RET and MN-NCE in both treatment and control groups under the two different administration conditions (P> 0.05). In addition, there was no significant difference in the apoptosis rate of spleen cells between the two groups (P>0.05),and there were no significant pathological changes in the spleen tissue. On the other hand, significant and positive results were observed with the cyclophosphamide groups. CONCLUSION: The micronucleus test results show that the biodegradable patches had no potential genotoxic risk. In addition,the selection and analysis of MN-RET and MN-NCE in mouse peripheral blood under different dosing cycle conditions was useful in detecting genotoxic risk of chemicals.

Key words: flow cytometry, micronucleus test, genotoxicity, biodegradable patches

中图分类号:

R994.6

王国伟, 孙晓霞, 车国喜, 盖潇潇, 屈秋锦, 王焱, 汪晓飞, 杜晓丹, 刘成虎. 外周血微核试验检测可降解生物材料补片的遗传毒性[J]. 癌变·畸变·突变, 2023, 35(5): 387-391.

WANG Guowei, SUN Xiaoxia, CHE Guoxi, GAI Xiaoxiao, QU Qiujin, WANG Yan, WANG Xiaofei, DU Xiaodan, LIU Chenghu. Genotoxicity evaluation of biodegradable patches using the micronucleus test in peripheral blood of mice[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2023, 35(5): 387-391.

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外周血微核试验检测可降解生物材料补片的遗传毒性

Genotoxicity evaluation of biodegradable patches using the micronucleus test in peripheral blood of mice

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