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JECFA also evaluated estimates of dietary exposure to titanium dioxide, estimating the maximum 95^th percentile of exposure to be 10 mg/kg BW per day. Overall, considering the low oral absorption of titanium dioxide as a food additive, the committee reaffirmed the ADI “not specified” that was established at the 13^th meeting.

A  2023 study published in the journal Environmental Research, scientists examined the effect of titanium dioxide nanoparticles on important gut bacteria in mice. Their results showed “the growth inhibitory effects could be associated with cell membrane damage caused by titanium dioxide nanoparticles to the bacterial strains. Metabolomics analysis showed that TiO2 NPs caused alterations in multiple metabolic pathways of gut bacteria, such as tryptophan and arginine metabolism, which were demonstrated to play crucial roles in regulating gut and host health.” The researchers also found that four different neuroprotective metabolites “were significantly reduced” in urine and in vitro bacteria and vivo urine samples. The researchers concluded: “Increasing evidence implies that the gut microbiome plays a profound role in regulating host metabolism. Our results illustrated that TiO2 NPs hindered the growth of four beneficial gut bacterial strains.”

As mentioned above, these oxide NPs are harmful in part because both anatase and rutile forms are semiconductors and produce ROS. Particularly, P25 kind has band-gap energies estimated of 3.2 and 3.0 eV, equivalent to radiation wavelengths of approximately 388 and 414 nm, respectively. Irradiation at these wavelengths or below produces a separation of charge, resulting in a hole in the valence band and a free electron in the conduction band, due to the electron movement from the valence to conduction bands. These hole–electron pairs generate ROS when they interact with H₂O or O₂ [43,44]. It was described that they can cause an increase in ROS levels after exposure to UV-visible light [45]. The NBT assay in the studied samples showed that bare P25TiO₂NPs produce a large amount of ROS, which is drastically reduced by functionalization with vitamin B2 (Fig. 5). This vitamin, also known as riboflavin, was discovered in 1872 as a yellow fluorescent pigment, [46] but its function as an essential vitamin for humans was established more than sixty years later, and its antioxidant capacity was not studied until the end of the XX century [47,48]. This antioxidant role in cells is partially explained because the glutathione reductase enzyme (GR) requires it for good functionality. This enzyme is the one in charge of the conversion of oxidized glutathione to its reduced form which acts as a powerful inner antioxidant and can quench the ROS [49,50]. The cost of this action is that the glutathione is converted to the oxidized form and needs to be recovered by the GR. Consequently, the cells need more vitamin B2. Another glutathione action is the protection against hydroperoxide. This activity is also mediated by riboflavin. Therefore, local delivery of this vitamin seems to significantly help the cells in their fight to keep the oxidative balance, once they are exposed to high levels of ROS.

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