Endocytosis of cerium oxide nanoparticles and modulation of reactive oxygen species in human ovarian and colon cancer cells

Cerium oxide nanoparticles (nanoceria) are widely reported to be cytocompatible and modulate intracellular reactive oxygen species (ROS) in a range of different cell types. In this study, nanoceria (d = 7 and 94 nm) synthesised by flame spray pyrolysis did not affect the proliferation of SKOV3 human ovarian and WiDr human colon cancer cell lines over a 72 h treatment period. The cellular accumulation of nanoceria was uniform and increased up to 24 h post-treatment before decreasing. The uptake of nanoceria in both cell lines was energy-dependent and was found to occur via non-specific pathways as well as clathrin-coated vesicles and caveolae. Nanoceria were localised predominantly in the cytoplasm and, to a lesser extent, with clathrin, caveolin-1 and lysosomes. The intracellular trafficking varied with particle size, treatment time and cell type. The larger nanoceria were found to scavenge intracellular ROS to a greater extent than the smaller nanoceria, and ROS scavenging was found to increase with treatment time. Together these data demonstrated that the diameter of the nanoceria and the cell types determined their mechanisms of uptake and intracellular localisation, as well as their ROS scavenging effects. Statement of Significance Cerium oxide nanoparticles (nanoceria) are a promising biomaterial that can catalytically scavenge reactive oxygen species (ROS). Modulation of ROS may potentially minimise the inflammatory effects of cancer. However, the antioxidant properties of nanoceria are reported to be pH-dependent and, thus, dependent on their mechanisms of endocytosis. This study is the first to examine the effects of particle size on the uptake and intracellular trafficking of flame spray-synthesised nanoceria in human cancer cells. This study demonstrated that the particle diameter, treatment time and cell type determined the mechanisms of uptake and intracellular localisation of nanoceria, as well as their ROS scavenging effects. This study highlighted the importance of testing new nanoparticle systems rather than making assumptions based on previous uptake studies.