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From studies deemed relevant, the experts found that titanium dioxide as a food additive is poorly absorbed by the gastrointestinal tract of mice and rats, with no adverse effects observed in short-term studies in rodents receiving titanium dioxide in their diets. No observed adverse effect levels (NOAELs) of 15,000 milligrams per kilogram of bodyweight (mg/kg BW) per day and 5,000 mg/kg BW per day—the highest doses tested—were established for mice and rats, respectively.

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The production of ROS was studied on white blood cells as a model to screen the effect on eukaryotic cells after being exposed to samples and solar simulated irradiation (according to the level of penetration under the skin). For that purpose, the leukocytes were separated from anticoagulated fresh blood using the Ficoll-Hypaque reactive in a well-known technique [33]. Then, 50 μL of suspensions of P25TiO2NPs (0.2 mg/mL and 0.02 mg/mL), vitaminB2@P25TiO2NPs (0.2 mg/mL and 0.02 mg/mL) and vitamin B2 (0.2 mg/mL and 0.02 mg/mL) were prepared and mixed with 50 μL of white blood cells suspension. A solution of 3% H2O2 was used as positive control and PBS as negative control. Then, the samples were irradiated using the LED panel for 3 and 6 h to simulate the light penetration into the skin. Also, a set of samples was kept in the dark as control. Finally, the ROS were detected through the colorimetric assay employing the nitroblue tetrazolium salt (NBT salt) and the absorbance at 650 nm was measured. The experiment was reproduced twice; the standard deviation was calculated and p-value < 0.05 were considered significant.

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