Cell Biology in Environmental Toxicology (CBET) Research Group, Dept. Zoology and Animal Cell Biology, Faculty of Science and Technology and Research Centre for Experimental Marine Biology and Biotechnology PIE-UPV/EHU, University of the Basque Country UPV/EHU, E-48080, Bilbao, Basque Country, Spain.
Department of Bioscience - Soil Fauna Ecology and Ecotoxicology, Vejlsøvej 25. Building M3.14, 8600, Silkeborg, Denmark.
Ecotoxicol Environ Saf. 2019 Nov 15;183:109545. doi: 10.1016/j.ecoenv.2019.109545. Epub 2019 Aug 22.
Earthworm immune cells (coelomocytes) have become a target system in ecotoxicology due to their sensitivity against a wide range of pollutants, including silver nanoparticles (AgNPs). Presently, in vitro approaches (viability assays in microplate, flow cytometry, cell sorting) with primary cultures of Eisenia fetida coelomocytes have been successfully used to test the toxicity and the dissimilar response of cell subpopulations (amoebocytes and eleocytes) after PVP-PEI coated AgNPs and AgNO exposures. In order to obtain reliable data and to accurately assess toxicity with coelomocytes, first an optimal culture medium and the most responsive assay were determined. AgNPs posed a gradual decrease in coelomocytes viability, establishing the LC value in RPMI-1640 medium at 6 mg/l and discarding that the observed cytotoxicity was attributable to its coating agent PVP-PEI. Exposure to AgNPs caused selective cytotoxicity in amoebocytes, which correlated with the Ag concentrations measured in sorted amoebocytes and reinforced the idea of dissimilar sensitivities among amoebocytes and eleocytes. Silver nano and ionic forms exerted similar toxicity in coelomocytes. The in vitro approaches with coelomocytes of E. fetida performed in this study have the capacity to predict impairments caused by pollutants at longer exposure levels and thus, provide rapid and valuable information for eco(nano)toxicology.
由于蚯蚓免疫细胞（体腔细胞）对包括银纳米颗粒（AgNPs）在内的多种污染物具有敏感性，因此它们已成为生态毒理学的目标系统。目前，已经成功地使用欧洲红蚯蚓体腔细胞的原代培养物的体外方法（微孔板中的活力测定、流式细胞术、细胞分选）来测试 PVP-PEI 涂层的 AgNPs 和 AgNO 暴露后细胞亚群（阿米巴细胞和白细胞）的毒性和不同反应。为了获得可靠的数据并准确评估体腔细胞的毒性，首先确定了最佳的培养基和最敏感的测定方法。AgNPs 逐渐降低体腔细胞的活力，在 RPMI-1640 培养基中确定 LC 值为 6mg/l，并排除观察到的细胞毒性归因于其涂层剂 PVP-PEI。AgNPs 对阿米巴细胞造成选择性细胞毒性，这与分选的阿米巴细胞中测量的 Ag 浓度相关，并强化了阿米巴细胞和白细胞之间敏感性不同的观点。纳米银和离子银形式在体腔细胞中表现出相似的毒性。本研究中使用欧洲红蚯蚓体腔细胞进行的体外方法能够预测在较长暴露水平下污染物造成的损伤，从而为生态（纳米）毒理学提供快速而有价值的信息。
