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Titanium dioxide (TiO 2 ) nanoparticles are promising biomedical agents characterized by good biocompatibility. In this study, we explored the cytotoxicity of TiO 2-x nanoparticles with a different Ti 3+ (Ti 2+ )/Ti 4+ ratio and analyzed the effeciency of eryptosis indices as a tool in nanotoxicology.Methods. Two types of TiO 2-x nanoparticles (5 nm) with various Ti 3+ (Ti 2+ )/ Ti 4+ ratios in the crystal lattice were synthesized. 1-TiO 2-x nanoparticles contained 54% Ti 4+ , 38% Ti 3+ and 8% Ti 2+ , while the relative amount of Ti 4+ and Ti 3+ in the crystal lattice of 2-TiO 2-x nanoparticles was 63% and 37%, respectively. Cell viability and cell motility induced by TiO 2-x nanoparticles were investigated on primary broblast cultures. Eryptosis modulation by the nanoparticles along with cell death mechanisms was studied on rat erythrocytes.Results. We report that both TiO 2-x nanoparticles don't decrease the viability of broblasts simultaneously stimulating cell migration. Data from in vitro studies on erythrocytes indicate that TiO 2-x nanoparticles trigger eryptosis via ROS-(1-TiO 2-x ) and Ca 2+ -mediated mechanisms (both TiO 2-x nanoparticles) suggesting that evaluation of eryptosis parameters is a more sensitive nanotoxicological approach for TiO 2-x nanoparticles than cultured broblast assays.Conclusion. TiO 2-x nanoparticles are characterized by low toxicity against broblasts, but they induce eryptosis, which is shown to be a promising tool for nanotoxicity screening. The Ti 3+ (Ti 2+ )/ Ti 4+ ratio at least partially determines the cytotoxicity mechanisms for TiO 2-x nanoparticles.

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