Kim Rosa, Heo Yunwi, Yoon Hakwon, Park June-Woo
Environmental Exposure and Toxicology Research Center, Korea Institute of Toxicology (KIT), Jinju, Republic of Korea.
Department of Ocean Integrated Science, Chonnam National University, Yeosu, Republic of Korea.
Front Toxicol. 2024 Nov 7;6:1476110. doi: 10.3389/ftox.2024.1476110. eCollection 2024.
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 () 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.
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-SiONPs). 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.
The FITC-SiONPs 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 LC values of several NMs were lower in dechorionated embryos than those in intact ones.
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.
为了满足评估纳米材料（NM）毒性以及符合“3R”原则（替代、减少和优化动物实验）的日益增长的需求，斑马鱼胚胎已成为研究NM毒性的一种有前景的替代模型。然而，斑马鱼胚胎被一层无细胞包膜——卵膜所包围，这限制了纳米材料的渗透性。本研究调查了去卵膜斑马鱼胚胎在评估NM毒性中的重要性。
我们利用共聚焦显微镜和场发射扫描电子显微镜结合能谱分析，使用50纳米的异硫氰酸荧光素掺入的二氧化硅纳米颗粒（FITC-SiONPs）观察纳米材料进入胚胎体的渗透性。我们研究了用链霉蛋白酶去除卵膜对斑马鱼胚胎的生理影响。根据斑马鱼胚胎中卵膜的有无，使用标准化方法ISO/TS 22082:2020比较纳米毒性。
FITC-SiONPs吸附在胚胎卵膜上；卵膜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卵膜胚胎。去卵膜斑马鱼胚胎未表现出负面生理影响。几种纳米材料在去卵膜胚胎中的半数致死浓度（LC）值低于完整胚胎。
去卵膜斑马鱼胚胎对纳米材料表现出比通常更高的敏感性。据我们所知，这是第一项使用新的标准化测试方法ISO/TS 22082:2020评估纳米毒性的研究，并且可能有助于提高去卵膜胚胎作为评估纳米毒性替代模型的实用性。
