In vitro, in the hemocytes of the marine mussel Mytilus hemocytes, suspension of TiO₂ NPs (Degussa P25, 10 μg/ml) stimulated immune and inflammatory responses, such as lysozyme release, oxidative burst and nitric oxide production. Vevers and Jha demonstrated the intrinsic genotoxic and cytotoxic potential of TiO₂ NPs on a fish-cell line derived from rainbow-trout gonadal tissue (RTG-2 cells) after 24 h of exposure to 50 μg/ml. Reeves et al. demonstrated a significant increase in the level of oxidative DNA damage in goldfish cells, and suggested that damage could not repaired by DNA repair mechanisms. Another suggestion from the mentioned study was that hydroxyl radicals are generated also in the absence of UV light. It has been shown that fish cells are generally more susceptible to toxic/oxidative injury than mammalian cells.

In 2021, the European Food Safety Authority concluded that titanium dioxide is no longer safe in foods due to the same concerns over nanoparticles. As a result, titanium dioxide is now banned as a food additive in the EU. Although studies have shown that the absorption of ingested titanium dioxide is low, evidence suggests that titanium dioxide nanoparticles can accumulate in the body over time. Health Canada deemed it safe in 2022 but noted concerns. Unlike their European counterparts, Canadian officials did not consider studies performed with titanium dioxide nanoparticles alone. 

One of the key challenges facing TiO2 industry suppliers is the fluctuation of raw material prices. The production of TiO2 requires raw materials such as ilmenite or rutile, which are subject to market price volatility. Suppliers must carefully monitor these fluctuations and adjust their production processes accordingly to ensure profitability.

The vitaminC@P25TiO₂NPs, on the other side, did not have any effect on cell protection against ROS. This might be due to the fact that vitamin C, a well-known scavenger of ROS, could behave as prooxidant and even promote ROS and lipid peroxidation [39]. It was recently described that at small concentrations of vitamin C, the prooxidant effects dominate; while in large concentrations the antioxidant ones predominate [40]. The effect also depends on the cell state and the interaction of vitamin C with light. In this case, ascorbic acid may act as an antenna to harvest visible light when conjugated to P25TiO₂NPs. Indeed, it was previously found that this combination (in some ratios) could have an improved photocatalytic activity, possibly due to a red shift in its light absorbance [41]. Further studies on vitaminC@P25TiO₂NPs were not conducted, because of the poor antioxidant capacity [42].

Amount: if too little titanium dioxide is added, the distance between particles is too large and there is not enough opacity, while if the titanium content is too high, the particles interfere with each other and poor efficiency is obtained.