In a 2020 study published in the Journal of Trace Elements in Medicine and Biology, researchers conducted an in vitro experiment to analyze the effects of TiO2 nanoparticles on a human neuroblastoma (SH-SY5Y) cell line. The scientists evaluated “reactive oxygen species (ROS) generation, apoptosis, cellular antioxidant response, endoplasmic reticulum stress and autophagy.” The results showed that exposure to the nanoparticles “induced ROS generation in a dose dependent manner, with values reaching up to 10 fold those of controls. Nrf2 nuclear localization and autophagy also increased in a dose dependent manner. Apoptosis increased by 4- to 10-fold compared to the control group, depending on the dose employed.” 

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.