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Lyophilized vitamins@P25TiO₂NPs were obtained through the described methods with excellent reproducibility and yield: over 99% of initial P25TiO₂NPs were functionalized.

No acute effects of nano-sized TiO₂ were observed in Danio rerio (zebrafish) embryos. Exposure of rainbow trout to TiO₂ NPs triggered lipid peroxidation, influence on the respiratory tract, disturbance in the metabolism of Cu and Zn, induction of intestinal erosion  and accumulation in kidney tissue. Linhua et al. exposed juvenile carp to 100 and 200 mg/ml of particles and TiO₂ observed no mortality. However, the fish suffered from oxidative stress and pathological changes in gill and liver. In the infaunal species Arenicola marina, exposure to TiO₂ NPs in sediment caused sub-lethal effects including decrease in casting rate and increase in cellular and DNA damage. Aggregated particles were visible in the lumen of the gut, but no uptake through the gut or the skin was observed.

Testing samples were made mixing 100 uL of TiO₂NPs suspensions (0.2 mg/mL and 0.02 mg/mL) and vitamins@P25TiO₂NPs (0.2 mg/mL and 0.02 mg/mL) with 100 μL ATCC 29,213 methicillin-sensitive Staphylococcus aureus (MSSA) (10⁷ in PBS, pH 7). Controls were made replacing nanoparticles with the same volume of PBS. The concentrations of nanoparticle suspensions were chosen based on the FDA approved maximal and the minimal amount usually found in sunscreens, which are 20% and 2% (this is equivalent to 0.2 mg/mL and 0.02 mg/mL for nanoparticles suspensions). The cream concentration, on the other hand, was an intermediate value of 10%.