Dechorionated zebrafish embryos improve evaluation of nanotoxicity.

IF 3.6 Q2 TOXICOLOGY Frontiers in toxicology Pub Date : 2024-11-07 eCollection Date: 2024-01-01 DOI:10.3389/ftox.2024.1476110
Rosa Kim, Yunwi Heo, Hakwon Yoon, June-Woo Park
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Abstract

Introduction: In response to the growing need to evaluate nanomaterial (NM) toxicity and compliance with the "3Rs" principles (replacement, reduction, and refinement of animal experiments), zebrafish (Danio rerio) embryos have emerged as a promising alternative model for studies on NM toxicity. However, zebrafish embryos are surrounded by an acellular envelope, the chorion, which limits the permeability of NMs. The present study investigated the importance of dechorionated zebrafish embryos for evaluating NM toxicity.

Methods: We utilized confocal microscopy and field-emission scanning electron microscopy with energy-dispersive spectroscopy to observe the permeability of NMs into the embryonic body using 50-nm fluorescein 5 (6)-isothiocyanate-incorporated silica nanoparticles (FITC-SiO2NPs). We investigated the physiological effects of removing the chorion using pronase on zebrafish embryos. Nanotoxicity was compared depending on the presence or absence of the chorion in zebrafish embryos using the standardized method ISO/TS 22082:2020.

Results: The FITC-SiO2NPs were adsorbed onto the embryonic chorion; the Si content was higher in the chorion than in the embryonic body and higher in the intact zebrafish embryos than in the dechorionated ones. Dechorionated zebrafish embryos exhibited no negative physiological effects. The LC50 values of several NMs were lower in dechorionated embryos than those in intact ones.

Conclusion: Dechorionated zebrafish embryos exhibited greater sensitivity to NMs than usual. To the best of our knowledge, this is the first study to evaluate NM toxicity using a new standardized test method, ISO/TS 22082:2020, and could contribute towards the increased utility of dechorionated embryos as an alternative model for the evaluation of nanotoxicity.

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去绒毛的斑马鱼胚胎改进了纳米毒性评估。
简介:为了满足评估纳米材料(NM)毒性和遵守 "3R "原则(替代、减少和改进动物实验)的日益增长的需求,斑马鱼(Danio rerio)胚胎已成为研究 NM 毒性的一种有前途的替代模型。然而,斑马鱼胚胎周围有一层无细胞包膜--绒毛膜,这限制了核磁的渗透性。本研究探讨了去绒毛斑马鱼胚胎对评估核磁共振毒性的重要性:方法:我们利用共聚焦显微镜和场发射扫描电子显微镜以及能量色散光谱仪,使用 50 纳米荧光素 5 (6) - 异硫氰酸酯并入的二氧化硅纳米粒子(FITC-SiO2NPs)观察 NMs 对胚胎的渗透性。我们研究了使用代孕酶去除绒毛对斑马鱼胚胎的生理影响。使用 ISO/TS 22082:2020 标准方法比较了斑马鱼胚胎中绒毛膜存在与否所产生的纳米毒性:结果:FITC-SiO2NPs 被吸附在胚胎绒毛上;绒毛中的硅含量高于胚胎体,完整斑马鱼胚胎中的硅含量高于去绒毛的斑马鱼胚胎。去绒毛的斑马鱼胚胎没有表现出负面的生理影响。结论:去绒毛斑马鱼胚胎中几种 NM 的 LC50 值低于完整斑马鱼胚胎:结论:脱绒毛斑马鱼胚胎对 NMs 的敏感性高于正常斑马鱼胚胎。据我们所知,这是首次使用新的标准化测试方法(ISO/TS 22082:2020)来评估 NM 毒性的研究,有助于提高去绒毛膜胚胎作为纳米毒性评估替代模型的实用性。
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