Assessment of Genotoxicity of Tissue Engineered Materials Based on Improved in vivo Hepatocyte Unscheduled DNA Synthesis(UDS)Assay
10.13241/j.cnki.pmb.2025.17.003
- VernacularTitle:基于改良体内肝细胞UDS试验的组织工程材料遗传毒性评估
- Author:
Luan-luan WANG
1
;
Li HOU
1
;
Xiang-yu CHU
1
;
Zi-yi YANG
1
;
Ling-xiao SUN
1
;
Xiao-fei WANG
1
;
Qiu-jin QU
1
;
Jing XU
1
;
Zeng-xiang LIU
1
;
Xiao-xia SUN
1
Author Information
1. 山东省医疗器械和药品包装检验研究院生物材料器械质量评价共性技术山东省工程研究中心 山东济南 250101
- Publication Type:Journal Article
- Keywords:
Genotoxicity;
Unscheduled DNA Synthesis Test;
5-ethynyl-2-deoxyuridine;
Tissue Engineered Materials
- From:
Progress in Modern Biomedicine
2025;25(17):2740-2748
- CountryChina
- Language:Chinese
-
Abstract:
Objective:An in vivo mammalian hepatocyte Unscheduled DNA Synthesis(UDS)test was used to evaluate the genotoxicity of Cross-linked Sodium Hyaluronate Gel and Bone Repair Materials,providing experimental evidence for establishing a UDS testing method for medical devices and materials.Methods:0.9％sodium chloride injection and cottonseed oil were used as the solvent for test materials and negative control,respectively.N-dimethylnitrosamine(NDMA)was used as the positive control for the early sampling times,and 2-acetylaminofluorene(2-AAF)was used as the positive control for the late sampling times.SD rats were administered a single dose for toxic exposure,and liver tissues were collected at 4 h and 16 h,respectively.Hepatocytes were isolated using collagenase perfusion.After labeling with 5-ethynyl-2'-deoxyuridine(EdU),and the net average fluorescence intensity(NAFI)of cell nuclei and nucleoplasm was measured by fluorescence microscope.Data from 50 cells were used to analyze the DNA repair level.Results:Compared with the negative control groups,the positive control groups(NDMA and 2-AAF)showed highly statistically significant differences in NAFI(P＜0.01),indicating successful induction of DNA damage.There was no statistically significant differences between the cross-linked sodium hyaluronate gel groups,bone repair material groups and the negative control group(P＞0.05),suggesting that these materials did not significantly induce DNA damage under the experimental conditions.Conclusion:This study first applied EdU labeling technology to the in vivo hepatic UDS assay,achieving non-radioactive labeling through click chemistry reactions.Under the conditions of this study,cross-linked sodium hyaluronate gel and bone repair materials did not exhibit genotoxicity.In the follow-up,the sample range can be expanded and the observation period can be prolonged to further improve the genotoxicity evaluation system of medical devices.
