Silica nanoparticles and silver-doped silica nanoparticles induce endoplasmatic reticulum stress response and alter cytochrome P4501A activity

Engineered silica nanoparticles (SiO2-NPs) find widespread application and may lead to exposure of humans and the environment. Here we compare the effects of SiO2-NPs and SiO2-NPs doped with silver (SiO2–Ag-NPs) on survival and cellular function of human liver cells (Huh7) and Pimephales promelas (fathead minnow) fibroblast cells (FMH). In Huh7 cells we investigate effects on the endoplasmatic reticulum (ER), including ER stress, and interactions of nanoparticles (NPs) with metabolizing enzymes and efflux transporters. The NPs formed agglomerates/aggregates in cell culture media as revealed by SEM and TEM. SiO2 and SiO2–1% Ag-NPs were taken up into cells as demonstrated by agglomerates occurring in vesicular-like structures or freely dispersed in the cytosol. Cytotoxicity was more pronounced in Huh7 than in FMH cells, and increased with silver content in silver-doped NPs. Dissolved silver was the most significant factor for cytotoxicity. At toxic and non-cytotoxic concentrations SiO2-NPs and SiO2–1% Ag-NPs induced perturbations in the function of ER. In Huh7 cells NPs induced the unfolded protein response (UPR), or ER stress response, as demonstrated in induced expression of BiP and splicing of XBP1 mRNA, two selective markers of ER stress. Additionally, SiO2–1% Ag-NPs and AgNO3 induced reactive oxygen species. Pre-treatment of Huh7 cells with SiO2–1% Ag-NPs followed by exposure to the inducer benzo(a)pyrene caused a significant reduced induction of CYP1A activity. NPs did not alter the activity of ABC transporters. These data demonstrate for the first time that SiO2-NPs and SiO2–1% Ag-NPs result in perturbations of the ER leading to the ER stress response. This represents a novel and significant cellular signalling pathway contributing to the cytotoxicity of NPs.

► Cellular effects of silica nanoparticles (SiO2-NPs) and silver-doped SiO2-NPs are assessed.
► Human liver cells Huh7 are more sensitive to cytotoxicity than fish cells.
► SiO2-NPs and silver-doped SiO2-NPs induce endoplasmatic reticulum (ER) stress response.
► ER stress response is a novel mode of action contributing to ER damage and adverse effects.
► NPs negatively interact with CYP1A induction and induce oxidative stress.