New Data on Variously Directed Dose-Response Relationships and the Combined Action Types for Different Outcomes of in Vitro Nanoparticle Cytotoxicity.

Spherical selenium-oxide and copper-oxide nanoparticles (SeO-NP with mean diameter 51 ± 14 nm and CuO-NP with mean diameter 21 ± 4 nm) were found to be cytotoxic for human fibroblast-like cells in vitro, as judged by decreased ATP-dependent luminescence. Compared with SeO-NP, CuO-NP produced a somewhat stronger effect of this kind. Along with cell hypertrophy developing in response to certain doses of SeO-NP and CuO-NP, our experiment also revealed doses causing a decrease in cell and cell-nucleus sizes. We observed both monotonic and different variants of nonmonotonic dose-response relationship. For the latter, we have succeeded in constructing adequate mathematical expressions based on the generalized hormesis paradigm that we had considered previously in respect of CdS-NP and PbS-NP cytotoxicity for cardiomyocites. It was demonstrated as well that combined toxicity of SeO-NP and CuO-NP is of different types depending on the outcome.

More data on in vitro assessment of comparative and combined toxicity of metal oxide nanoparticles.

Isolated and combined damaging effects of PbO and CuO nanoparticles were estimated on an established line of human fibroblasts by a decrease in: (a) the cellular dehydrogenase activity (MTT Assay), (b) the ATP content (Luminescent Cell Viability Assay), (c) the cellular proliferation, viability, spreading, and attachment to substrate evaluated integrally by continuous impedance-based measurement of the Normalized Cell Index. Using all these indices, we demonstrate an explicit dependence of cell damage on the concentrations of both metal oxide nanoparticle (MeO-NP) species. This dependence is adequately approximated with a hyperbolic function. At equal exposure levels, PbO-NP and CuO-NP demonstrate quantitatively similar cytotoxicities. The same was observed previously for some non-specific in vivo toxicity measures. The combined in vitro cytotoxicity has also been described mathematically using the Response Surface Methodology and found to be represented by various types, thus corroborating, in this respect also, the findings of a previous animal experiment with the same MeO-NPs.

Are in vivo and in vitro assessments of comparative and combined toxicity of the same metallic nanoparticles compatible, or contradictory, or both? A juxtaposition of data obtained in respective experiments with NiO and Mn3O4 nanoparticles.

Comparative and combined damaging effects of NiO and Mn3O4 nanoparticles were estimated on cultures of several established human cell lines. The cytotoxicity indices used were: (a) reduction in cellular dehydrogenase activity, (b) decrease in the ATP-content, (c) for SH-SY5Y cells also decrease in the tyrosine hydroxylase content. The combined cytotoxicity was modeled using the Response Surface Methodology. When assessing the stability of metal oxide nanoparticles (MeO-NPs) in cultural media used by us, we found that the addition of the fetal bovine serum (FBS) to them renders NiO-NPs and, to even greater extent, Mn3O4-NPs exponentially slow soluble while without FBS their dissolution was virtually undetectable. At the same time, sedimentation of these MeO-NPs noticeably slowed down in the presence of the same FBS. We have found dependence of cell damage on concentrations of MeO-NPs and higher cytotoxicity of Mn3O4-NP compared with NiO-NP. Thus, comparative assessment of the NPs unspecific toxicity obtained in our animal experiments was reproduced by the "in vitro" tests. However, with respect to manganese-specific brain damage "in vivo" discovered previously, present experiments on neurons "in vitro" showed only a certain enhancing effect of Mn3O4-NP on the action of NiO-NP, but the role of NiO-NP in the combination prevailed.