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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.

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In addition to the toxic effects of TiO₂ NPs, discussed in previous chapters, these NPs have been also shown to promote photosynthesis and nitrogen metabolism, resulting in the enhanced growth of spinach. It increases the absorption of light and accelerates the transfer and transformation of the light energy. It was also found that treatment with nano-sized TiO₂ significantly increased the level of antioxidant enzymes, and decreased the ROS accumulation and malonyldialdehyde content in spinach chloroplasts under visible and UV irradiation. TiO₂ NPs also increased the superoxide dismutase activity of germinating soybean, enhanced its antioxidant ability, and promoted seed germination and seedling growth.