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 TiO2, 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.
≥ 5 % of standard sample
Testing samples were made mixing 100 uL of TiO2NPs suspensions (0.2 mg/mL and 0.02 mg/mL) and vitamins@P25TiO2NPs (0.2 mg/mL and 0.02 mg/mL) with 100 μL ATCC 29,213 methicillin-sensitive Staphylococcus aureus (MSSA) (107 in PBS, pH 7). Controls were made replacing nanoparticles with the same volume of PBS. The concentrations of nanoparticle suspensions were chosen based on the FDA approved maximal and the minimal amount usually found in sunscreens, which are 20% and 2% (this is equivalent to 0.2 mg/mL and 0.02 mg/mL for nanoparticles suspensions). The cream concentration, on the other hand, was an intermediate value of 10%.
In order to achieve the same solids content, the larger filler and the binder should be reduced if necessary.