Jimeno-Romero Alba, Gwinner Frederik, Müller Michelle, Mariussen Espen, Soto Manu, Kohl Yvonne
Fraunhofer Institute for Biomedical Engineering IBMT, 66280 Sulzbach, Saar, Germany.
Cell Biology and Environmental Toxicology Group, Research Centre for Experimental Marine Biology and Biotechnology, University of the Basque Country UPV/EHU, 48620 Plentzia, Basque Country, Spain.
Nanomaterials (Basel). 2021 Nov 20;11(11):3136. doi: 10.3390/nano11113136.
Determination of acute toxicity to vertebrates in aquatic environments is mainly performed following OECD test guideline 203, requiring the use of a large number of fish and with mortality as endpoint. This test is also used to determine toxicity of nanomaterials in aquatic environments. Since a replacement method for animal testing in nanotoxicity studies is desirable, the feasibility of fish primary cultures or cell lines as a model for nanotoxicity screenings is investigated here. primary cultures and RTgill-W1 cell line were exposed to several concentrations (0.1 to 200 ug/mL) of different nanoparticles (TiO, polystyrene and silver), and cytotoxicity, metabolic activity and reactive oxygen species formation were investigated after 24 and 48 h of exposure. Protein corona as amount of protein bound, as well as the influence of surface modification (-COOH, -NH), exposure media (Leibovitz's L15 or seawater), weathering and cell type were the experimental variables included to test their influence on the results of the assays. Data from all scenarios was split based on the significance each experimental variable had in the result of the cytotoxicity tests, in an exploratory approach that allows for better understanding of the determining factors affecting toxicity. Data shows that more variables significantly influenced the outcome of toxicity tests when the primary cultures were exposed to the different nanoparticles. Toxicity tests performed in RTgill-W1 were influenced only by exposure time and nanoparticle concentration. The whole data set was integrated in a biological response index to show the overall impact of nanoparticle exposures.
水生环境中对脊椎动物急性毒性的测定主要按照经合组织（OECD）测试指南203进行，该指南要求使用大量鱼类，并以死亡率作为终点指标。此测试也用于测定纳米材料在水生环境中的毒性。由于纳米毒性研究中需要动物试验的替代方法，因此本文研究了鱼类原代培养物或细胞系作为纳米毒性筛选模型的可行性。将原代培养物和RTgill-W1细胞系暴露于不同纳米颗粒（二氧化钛、聚苯乙烯和银）的几种浓度（0.1至200微克/毫升）下，在暴露24小时和48小时后研究细胞毒性、代谢活性和活性氧的形成。作为结合蛋白量的蛋白质冠层，以及表面修饰（-COOH、-NH）、暴露介质（莱博维茨L15培养基或海水）、老化和细胞类型的影响，均作为实验变量，以测试它们对检测结果的影响。根据每个实验变量在细胞毒性测试结果中的显著性，对所有情况的数据进行分类，采用探索性方法以便更好地理解影响毒性的决定因素。数据表明，当原代培养物暴露于不同纳米颗粒时，更多变量对毒性测试结果有显著影响。在RTgill-W1细胞系中进行的毒性测试仅受暴露时间和纳米颗粒浓度的影响。将整个数据集整合到一个生物反应指数中，以显示纳米颗粒暴露的总体影响。
