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The vitaminC@P25TiO2NPs, on the other hand, were obtained through an optimized method based on Mallakpour et al. [27]. Initially, 0.02 g of P25TiO2NPs were dispersed in 1 mL of ultrapure water and stirred in a Vortex. Next, 100 μL of HCl (0.01 M) were added (pH 2) to 100 uL of P25TiO2NPs to avoid gel formation. Then, 100 μL of vitamin C dissolved in ultra-pure water (5.0 × 10−3 M) solution were added to the mixture and was ultrasonicated for 30 min. Finally, vitamin C was added in excess to gain a beige-orange color suspension, and the ultrasonication continued for another 30 min. The pellet obtained after centrifuging the suspension for 10 min at 4500 rpm was resuspended in ultrapure water, centrifuged again, and then lyophilized.

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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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