Titanium is a metal element found naturally in the environment. When it's exposed to oxygen in the air, it forms titanium oxides that are contained in many minerals, sands, soils, and dusts.

Even though it is one of the most-produced chemicals, the real and potential benefits of titanium dioxide are not without controversies. Dust inhalation may cause breathing problems. Titanium dioxide has been classified by the International Agency for Research on Cancer as an Group 2B carcinogen, a “possible carcinogen to humans,” based on studies of rats that inhaled the substance.

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.