| Summary: | The aim of the present study was to investigate the eco-cytotoxicity of several forms of nanomaterials (NM), such as nano-CuO, nano-TiO<sub>2</sub>, nano-SiO<sub>2</sub> and nano-ZnO, on different aquatic species (<i>Raphidocelis subcapitata</i>, <i>Daphnia magna</i> and <i>Lemna minor</i>) following standard protocols and on human cell lines (Caco-2, SV-80, HepG2 and HaCaT). Predicted no-effect concentrations (PNEC) or hazard concentrations for 5% of the species (HC<sub>5</sub>) were also estimated based on the compilation of data available in the literature. Most of the NM agglomerated strongly in the selected culture media. For the ecotoxicity assays, nano-CuO and nano-ZnO even in particle agglomeration state were the most toxic NM to the freshwater organisms compared to nano-TiO<sub>2</sub> and nano-SiO<sub>2</sub>. Nano-ZnO was the most toxic NM to <i>R. subcapitata</i> and <i>D. magna</i>, while nano-CuO was found to be very toxic to <i>L. minor</i>. Nano-CuO was very toxic to Caco-2 and HepG2 cells, particularly at the highest tested concentrations, while the other NM showed no toxicity to the different cell lines. The HC<sub>5</sub> and PNEC values are still highly protective, due to data limitations. However, the present study provides consistent evidence of the potential risks of both nano-CuO and nano-ZnO against aquatic organisms and also their effects on public health.
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