Silver bionanoparticles toxicity in trophoblast is mediated by nitric oxide and glutathione pathways

Silver bionanoparticles (AgNPs) biosynthesized by Pseudomonas aeruginosa culture supernatant have an important antibacterial activity mediated by ROS increase; however their toxicity in human cells is not known. Due to the high susceptibility of the developing tissues to xenobiotics, the aim of this study was to investigate the AgNPs effect on first trimester human trophoblasts. The HTR8/SVneo cell line was treated with AgNPs (0.3-1.5 pM), for 6 and 24 h. Cell viability, reactive nitrogen and oxygen species (RNS and ROS) production, nitric oxide synthase expression, antioxidant defenses and biomolecule damage were evaluated. The exposure to AgNPs produced changes in HTR8/SVneo cell morphology and decreased cell viability. Alterations in redox balance were observed, with an increase in ROS and RNS levels, and NOS2 protein expression. Superoxide dismutase and catalase augmented their activity accompanied with a decreased in glutathione content and glutathione S-transferase activity. Protein oxidation and genotoxic damage were observed at concentrations greater than 0.6 pM. The pre-incubation with l-NMMA, NAC, mannitol and peroxidase demonstrated that AgNPs-induced cytotoxicity was not mediated by HO and H₂O₂, but nitric oxide and glutathione pathways were implicated in cell death. Since reported AgNPs microbicidal mechanism is mediated by increasing ROS (mainly HO and H₂O₂) without an increase in RNS, this work indicates an interesting difference in the reactive species and oxidative pathways involved in AgNPs toxicity in eukaryotic and prokaryotic cells. Highlighting the importance of toxicity evaluation to determine the safety of AgNPs with pharmaceutical potential uses.