Different dermal cell types have been reported to differ in their sensitivity to nano-sized TiO₂ . Kiss et al. exposed human keratinocytes (HaCaT), human dermal fibroblast cells, sebaceous gland cells (SZ95) and primary human melanocytes to 9 nm-sized TiO₂ particles at concentrations from 0.15 to 15 μg/cm² for up to 4 days. The particles were detected in the cytoplasm and perinuclear region in fibroblasts and melanocytes, but not in kerati-nocytes or sebaceous cells. The uptake was associated with an increase in the intracellular Ca²⁺ concentration. A dose- and time-dependent decrease in cell proliferation was evident in all cell types, whereas in fibroblasts an increase in cell death via apoptosis has also been observed. Anatase TiO₂ in 20–100 nm-sized form has been shown to be cytotoxic in mouse L929 fibroblasts. The decrease in cell viability was associated with an increase in the production of ROS and the depletion of glutathione. The particles were internalized and detected within lysosomes. In human keratinocytes exposed for 24 h to non-illuminated, 7 nm-sized anatase TiO_2, a cluster analysis of the gene expression revealed that genes involved in the “inflammatory response” and “cell adhesion”, but not those involved in “oxidative stress” and “apoptosis”, were up-regulated. The results suggest that non-illuminated TiO₂ particles have no significant impact on ROS-associated oxidative damage, but affect the cell-matrix adhesion in keratinocytes in extracellular matrix remodelling. In human keratinocytes, Kocbek et al. investigated the adverse effects of 25 nm-sized anatase TiO₂ (5 and 10 μg/ml) after 3 months of exposure and found no changes in the cell growth and morphology, mitochondrial function and cell cycle distribution. The only change was a larger number of nanotubular intracellular connections in TiO₂-exposed cells compared to non-exposed cells. Although the authors proposed that this change may indicate a cellular transformation, the significance of this finding is not clear. On the other hand, Dunford et al. studied the genotoxicity of UV-irradiated TiO₂ extracted from sunscreen lotions, and reported severe damage to plasmid and nuclear DNA in human fibroblasts. Manitol (antioxidant) prevented DNA damage, implying that the genotoxicity was mediated by ROS.

Both P25TiO₂NPs (with or without vitamin B2) were not found beyond the epidermis in 99% of the analyzed TEM images (Fig. 8). This is coherent with previous findings showing that nanoparticles greater than 50 nm can not penetrate the skin, even in vivo models with movement, stretching, and friction [54]. However, in one of the zones, a few nanoparticles were observed inside a hair follicle. This could be due to the follicle exposure after the localized rupture of this physical barrier when rats were shaved in order to clean the area for cream topical administration. This finding suggests that nanoparticle-based sunscreen should not be applied on recently shaved or harmed skin, in order to avoid nanoparticle skin penetration.

Although barium sulfate is almost completely inert, zinc sulfide degrades upon exposure to UV light, leading to darkening of the pigment. The severity of this UV reaction is dependent on a combination of two factors; how much zinc sulfide makes up the pigments formulation, and its total accumulated UV exposure. Depending on these factors the pigment itself can vary in shade over time, ranging from pure white all the way to grey or even black. To suppress this effect, a dopant may be used, such as a small amount of cobalt salts, which would be added to the formulation. This process creates cobalt-doped zinc sulfide. The cobalt salts help to stabilize zinc sulfide so it will not have as severe a reaction to UV exposure.