In order to contribute with experimental evidence that could help to achieve a better understanding of the field for future regulation, in the present work, the biocompatibility of commercial P25TiO₂NPs (one type of TiO₂NPs used in sunscreen formulations) and two novel functionalized P25TiO₂NPs were evaluated under solar simulated irradiation. White light, generated by red, blue, and yellow LEDs, together with UV ones, was chosen to simulate the solar spectra. Functionalization of TiO₂NPs was made with antioxidant vitamins in order to prevent the expected photo-initiated ROS production when nanoparticles are exposed to the simulated solar spectra. Vitamin B2 (riboflavin) and vitamin C were chosen to carry out the functionalization because they are water-soluble, low-cost, and are a constitutive part of biological processes. In addition, it is known that both have the potential to prevent macromolecular oxidation by ROS [23], [24], [25], [26].

Hemolysis was studied on 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 500 μL of anticoagulated blood (donated by Laboratorio de Hemoderivados, UNC) in a rate of 1/10. A solution of NaCl 10% was used as the positive control and PBS as the negative control. Then, the samples were irradiated using the LED described above 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 samples were centrifuged and the absorbance at 540 nm was measured in the supernatants. The experiment was reproduced twice; the standard deviation was calculated and p-value < 0.05 were considered significant.

Most notably, a European Food Safety Authority safety assessment published in May 2021 pointed to genotoxicity concerns, as suggested by previous research. Genotoxicity is the ability of chemicals to damage genetic information such as DNA, which may lead to cancer.