Fate and Toxicity of Engineered Inorganic Nanoparticles

Engineered nanomaterials are increasingly used in a variety of industrial processes and consumer products. Numerous studies have reported toxicity of different NPs during the last years. Thus, there are growing concerns about the potential impacts to the health and environment of engineered nanoparticles (NPs). However, some methodological problems complicate the interpretation of nanotoxicity studies. On the one hand, some NPs have shown to interfere with classical toxicity assays based on colorimetric or fluorescent measurements. On the other hand, most NPs tend to aggregate in media used in toxicity tests, which complicates the interpretation of the toxicity results. The first objective of this dissertation was to evaluate a novel impedance-based and label-free real time cell analyzer (RTCA) as a high throughput method for screening the cytotoxicity of nanoparticles and to validate the RTCA results using a conventional cytotoxicity test (MTT). Several inorganic NPs were tested for potential cytotoxicity to human bronchial epithelial cells (16HBE14o-). In general, there was a good correlation in cytotoxicity measurements between the two methods. Moreover, none of the NPs tested showed interference with the impedance measurements performed by the RTCA system. The results demonstrate the potential and validity of the impedance-based RTCA technique to rapidly screen for NP toxicity. The second objective of this dissertation was to assess the toxicity of different inorganic NPs to the eukaryotic cell model Saccharomyces cerevisiae, and to test the influence of NP aggregation state in their toxicity. Nanotoxicity was assessed by monitoring oxygen consumption in batch cultures and by analysis of cell membrane integrity. Mn₂O₃ NPs showed the highest inhibition of O₂ consumption and cell membrane damage, while the other NPs caused low or no toxicity to the yeast. Most NPs showed high tendency to aggregate in the assay medium, so a non-toxic dispersant was used to improve