The toxicity of P25TiO₂NPs was evaluated in both prokaryotic (Fig. 3) and eukaryotic cells (Fig. 4). The XTT assay was chosen to measure the cell viability in bacterial cultures of MSSA, a normal skin microbiota microorganism. The reduction in the viability of samples with bare NPs is notorious, possibly due to the described ROS production from the interaction of P25TiO₂NPs with light [37]. This effect seems to be avoided when they are functionalized with vitamin B2. Also, the most concentrated vitaminB2@P25TiO₂NPs sample (0.2 mg/mL) shows up to 60% more absorbance after 6 h compared to the bare NPs (due to normal cell replication). This may indicate that the antioxidant effect of the vitamin B2 coating is greater than the oxidation damage produced by the NPs. This protective capacity could be attributed to the glutathione redox cycle and the conversion of reduced riboflavin to its oxidized form [38]. Values of cell viability greater than 100% are not rare and could be understood because the XTT assay actually measure metabolic activity when reducing the tetrazole to formazan. It is usually assumed that conversion is dependent on the number of viable cells, but it could also be related to an expected increased enzymatic activity when cells are exposed to small doses of some new substance. Further analysis showed that this effect was not the only one responsible for better cell viability of vitaminB@P25TiO₂NPs treated samples.

Freshwater algae show low-to-moderate susceptibility to TiO₂ exposure, with more pronounced toxic effects in the presence of UV irradiation. It has also been shown that nano-sized TiO₂ is significantly more toxic to algae Pseudokirchneriella sub-capitata than submicron-sized TiO₂. Hund-Rinke and Simon  reported that UV irradiated 25 nm TiO₂ NPs are more toxic to green freshwater algae Desmodesmus subspicatus than UV irradiated 50 nm particles, which is in agreement with Hartmann et al. UV irradiated TiO₂ NPs also inactivated other algae species such as Anabaena, Microcystis, Melsoira and Chroococcus. It was demonstrated that smaller particles have a greater potential to penetrate the cell interior than submicron-sized particles and larger aggregates. Studies have shown that the amount of TiO₂ adsorbed on algal cells can be up to 2.3 times their own weight.

Algaecidal effect of Lithopon: After 5 years of exposure to weathering in Alpen (Lower Rhine)  

The agency makes this exception for several approved color additives.