Novel dose metric for apparent cytotoxicity effects generated by in vitro cell exposure to silica nanoparticles

This study aimed at identifying the dose metric applicable to studies on the viability of cells exposed to nanoparticles (NPs) in vitro. A previously reported set of data was evaluated very carefully. The extent of cell death after 24-h exposure of three cell lines to suspended silica NPs (<30 nm) was quantified using four different viability/cytotoxicity assays. Data on NP uptake in cells after 6-h exposure were also reported. Evidence is provided that, in spite of the small size of the NPs, mass transport to the cells cannot be explained solely by diffusion. Gravitational settling must have contributed significantly, presumably as the result of the formation of large agglomerates. Appropriately adjusted response data, with typically 22 combinations of mass concentration and height of the medium for each cell line, could be integrated in universal diagrams, provided the dose was quoted in terms of the areal density of NP mass delivered to the cells. Loss of viability became observable only if cells were exposed to the equivalent of 1 to 5 closely packed layers of NPs; the dose required for complete cell death ranged between 4 and about 20 layers of NPs. The results suggest that the cell-death phenomena observed in the evaluated work and in many similar studies reported in the literature constitute a matter of cell overload with nanostructured matter. This finding also implies that the toxic potential of individual silicate NPs is very low. Strategies for the design of advanced future work are outlined.