French Agency for Food, Environmental and Occupational Health & Safety (ANSES), Fougères Laboratory, Toxicology of Contaminants Unit, Fougères, France.
French Agency for Food, Environmental and Occupational Health & Safety (ANSES), Fougères Laboratory, Toxicology of Contaminants Unit, Fougères, France.
Chemosphere. 2024 Feb;350:140975. doi: 10.1016/j.chemosphere.2023.140975. Epub 2023 Dec 22.
Nanomaterials (NMs) are defined as materials with at least one external dimension below 100 nm. Their small size confers them interesting unique physico-chemical properties, hence NMs are increasingly used in a diversity of applications. However, the specific properties of NMs could also make them more harmful than their bulk counterparts. Therefore, there is a crucial need to deliver efficient NM hazard assessment in order to sustain the responsible development of nanotechnology. This study analysed the genotoxic potential of several NMs: one titanium dioxide (TiO) and two zinc oxide NMs (ZnO) that were tested up to 100 μg/mL on 2D and 3D hepatic HepaRG models. Genotoxicity analysis was performed comparing the alkaline comet assay in classical and high throughput formats. Moreover, oxidative DNA lesions were investigated with the Fpg-modified comet assay. Results showed that TiO NMs were not cytotoxic and not genotoxic in either cell model, although a small increase in the % tail DNA was observed in 3D HepaRG cells at 100 μg/mL in the classical format. The two ZnO NMs (ZnO S. NMs a commercial suspension and NM110 provided by the European Union Joint Research Centre) induced a concentration-dependent increase in cytotoxicity that was more pronounced in the 2D (>20% cytotoxicity was observed for ZnO S. at concentrations greater than 25 μg/mL, and for NM 110 at 50 μg/mL) than in the 3D model (more than 20% cytotoxicity for ZnO S. NMs at 50 μg/mL). While ZnO S. NMs induced DNA damage associated with cytotoxicity (at 25 and 50 μg/mL in 2D and 50 μg/mL in 3D), NM110 showed a clear genotoxic effect at non-cytotoxic concentrations (25 μg/mL in 2D and at 25 and 50 μg/mL in 3D). No major differences could be observed in the comet assay in the presence or absence of the Fpg enzyme. High throughput analysis using CometChip® mostly confirmed the results obtained with the classical format, and even enhanced the detection of genotoxicity in the 3D model. In conclusion, this study demonstrated that new approach methodologies (NAMs), 3D models and the high throughput format for the comet assay, were more efficient in the detection of genotoxic effects, and are therefore promising approaches to improve hazard assessment of NMs.
纳米材料（NMs）被定义为至少有一个外部尺寸小于 100nm 的材料。它们的小尺寸赋予了它们有趣的独特物理化学性质，因此 NMs 越来越多地应用于各种应用中。然而，NMs 的特殊性质也可能使它们比其块状对应物更具危害性。因此，迫切需要进行有效的 NM 危害评估，以维持纳米技术的负责任发展。本研究分析了几种纳米材料的遗传毒性潜力：一种二氧化钛（TiO）和两种氧化锌纳米材料（ZnO），在二维和三维肝 HepaRG 模型中测试了高达 100μg/mL。通过经典和高通量格式的碱性彗星试验比较进行遗传毒性分析。此外，还使用 Fpg 修饰的彗星试验研究了氧化 DNA 损伤。结果表明，TiO NMs 在两种细胞模型中均无细胞毒性和遗传毒性，尽管在经典格式下，3D HepaRG 细胞在 100μg/mL 时观察到 %尾 DNA 略有增加。两种 ZnO NMs（ZnO S. 由欧盟联合研究中心提供的商业悬浮液和 NM110）诱导了浓度依赖性的细胞毒性增加，在 2D 中更为明显（对于 ZnO S.，浓度大于 25μg/mL 时观察到超过 20%的细胞毒性，对于 NM110，浓度大于 50μg/mL 时观察到超过 20%的细胞毒性），而在 3D 模型中则不明显（对于 ZnO S. NMs，浓度大于 50μg/mL 时观察到超过 20%的细胞毒性）。虽然 ZnO S. NMs 诱导了与细胞毒性相关的 DNA 损伤（在 2D 中为 25 和 50μg/mL，在 3D 中为 50μg/mL），但 NM110 在非细胞毒性浓度下表现出明显的遗传毒性作用（在 2D 中为 25μg/mL，在 2D 和 50μg/mL 中为 3D）。在存在或不存在 Fpg 酶的情况下，彗星试验的高通量分析大多证实了经典格式的结果，甚至增强了对 3D 模型中遗传毒性的检测。总之，本研究表明，新方法方法（NAMs）、3D 模型和彗星试验的高通量格式在检测遗传毒性方面更有效，因此是改进 NM 危害评估的有前途的方法。
Toxicol In Vitro. 2022-2
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