Silver nanoparticles induce oxidative stress, apoptosis and impaired steroidogenesis in ovarian granulosa cells of cattle.

There have been increased effects of silver nanoparticle (Ag-NPs) on livestock during the past decade, but data related to adverse effects of Ag-NPs on reproductive tissues are limited. In the present study, the possible cytotoxic effects of Ag-NPs on oxidant/antioxidant balance, apoptosis and steroid hormone production in ovarian granulosa cells of cattle were studied.Cultured granulosa cells were treated with 10 nm Ag-NPs at various concentrations (1-100 µg/ml) for 24 h, and cell toxicity, oxidant/antioxidant markers, reactive oxygen species (ROS) production, abundances of apoptotic/antiapoptotic and steroidogenesis related mRNA transcripts were determined. The amount of DNA fragmentation was also determined using the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Results indicated treatment of granulosa cells with Ag-NPs induced an increase of reactive oxygen species production concomitant with increased malondialdehyde concentrations and decreased antioxidant enzyme activities. There was a maximal percentage of TUNEL+ cells (46.6%) after treatment with 50 and 100 µg/ml of Ag-NPs. The Ag-NPs could induce apoptosis in granulosa cells as indicated by the increase in caspase-3 activity, and larger abundance of BCL2 associated X (BAX) and lesser abundance of B-cell lymphoma 2 (BCL2) mRNA transcripts. The abundance of steroidogenic enzyme mRNA transcripts decreased concomitant with suppression of steroid hormone synthesis from Ag-NP-treated cells. Findings indicate silver nanoparticles (SNP) induce apoptosis and oxidative stress and change the pattern of steroid hormone synthesis in granulosa cells of cattle. The results indicate Ag-NPs may inhibit the function and viability of ovarian cells.