The knowledge on potential harmful effects of metallic nanomaterials lags behind their increased use in consumer products and therefore, the safety data on various nanomaterials applicable for risk assessment are urgently needed. In this study, 11 metal oxide nanoparticles (MeOx NPs) prepared using flame pyrolysis method were analyzed for their toxicity against human alveolar epithelial cells A549, human epithelial colorectal cells Caco2 and murine fibroblast cell line Balb/c 3T3. The cell lines were exposed for 24 h to suspensions of 3-100 μg/mL MeOx NPs and cellular viability was evaluated using. Neutral Red Uptake (NRU) assay. In parallel to NPs, toxicity of soluble salts of respective metals was analyzed, to reveal the possible cellular effects of metal ions shedding from the NPs. The potency of MeOx to produce reactive oxygen species was evaluated in the cell-free assay. The used three cell lines showed comparable toxicity responses to NPs and their metal ion counterparts in the current test setting. Six MeOx NPs (Al2O3, Fe3O4, MgO, SiO2, TiO2, WO3) did not show toxic effects below 100 µg/mL. For five MeOx NPs, the averaged 24 h IC50 values for the three mammalian cell lines were 16.4 µg/mL for CuO, 22.4 µg/mL for ZnO, 57.3 µg/mL for Sb2O3, 132.3 µg/mL for Mn3O4 and 129 µg/mL for Co3O4. Comparison of the dissolution level of MeOx and the toxicity of soluble salts allowed to conclude that the toxicity of CuO, ZnO and Sb2O3 NPs was driven by release of metal ions. The toxic effects of Mn3O4 and Co3O4 could be attributed to the ROS-inducing ability of these NPs. All the NPs were internalized by the cells according to light microscopy studies but also proven by TEM, and internalization of Co3O4 NPs seemed to be most prominent in this aspect. In conclusion, this work provides valuable toxicological data for a library of 11 MeOx NPs. Combining the knowledge on toxic or non-toxic nature of nanomaterials may be used for safe-by-design approach.

Heavy metals are found naturally on Earth and exposure to them in the living environment is increasing as a consequence of human activity. The toxicity of six different metal oxide nanoparticles (NP) at different points in time was compared using resazurin assay. After incubating Caco2 and A549 cells with 100 lg/mL of Sb 2 O 3 , Mn 3 O 4 and TiO 2 nanoparticles (NPs) for 24 h no toxic effects were observed while Co 3 O 4 and ZnO NPs had moderate effects and CuO NPs were toxic below 100 lg/mL (24 h EC 25 = 11 for A549 and 71 lg/mL for Caco2). The long-term monitoring (up to 9 days) of cells to NPs revealed that the toxic effects of Mn 3 O 4 and Sb 2 O 3 NPs remarkably increased over time. The 9 day EC 50 values for Sb 2 O 3 NPs were 22 and 48 lg/mL for A549 and Caco2 cells; and for Mn 3 O 4 NPs were 47 and 29 lg/mL for A549 and Caco2 cells, respectively. In general, the sensitivity of the cell lines in the resazurin assay was comparable. Trans epithelial electrical resistance (TEER) measurements were performed for both cell types exposed to Co 3 O 4 , Sb 2 O 3 and CuO NPs. In TEER assay, the Caco2 cells were more susceptible to the toxic effects of these NPs than A549 cells, where the most toxic NPs were the Sb 2 O 3 NPs: the permeability of the Caco2 cell layer exposed to 10 lg/mL Sb 2 O 3 NPs already increased after 24 h of exposure.Electronic supplementary material The online version of this article

The use of phospholipid vesicles (liposomes) in EKC and CEC, as well as the use of CE for studying liposomes and lipid-analyte aggregates, are briefly reviewed. The works are presented in the order of appearance. The review describes the most common liposome dispersions used for achieving separation of neutral and charged compounds in liposome EKC. The CEC part is divided into two parts, discriminating between liposomes immobilized on capillaries for hindering analytes from interacting with the fused-silica wall and liposomes immobilized on capillaries for achieving chromatographic phases for separation of compounds. Finally, the use of CE for studying liposomes and lipid-analyte aggregates as analytes is discussed in brief.

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