Proceedings of The Physiological Society

Europhysiology 2018 (London, UK) (2018) Proc Physiol Soc 41, PCB197

Poster Communications

Cellular Effects of Silver Nanoparticles (AgNPs) on Human Keratinocyte Cell Line (HaCaT) in Vitro

K. Habas1, P. Asquith1, L. Shang1

1. Faculty of Life Sciences, University of Bradford, Bradford, United Kingdom.

Due to their intensive antibacterial properties, AgNPs have been widely used in many commercially products designed to direct contact with the skin [1]. This brings a growing concern on their pothential risks to human health [2]. Previous studies show that AgNPs can induce oxidative damage in the primary HaCaT cells responded to modulation of antioxidant capacity. AgNPs can generate elevated intracellular reactive oxygen species (ROS) and down-regulation of antioxidant enzymes such as GPX and SOD, resulting in the formation of DNA adducts [3]. The mechanisms involved in the endogenous antioxidants effect appear to be primarily independent on increased ROS production, but superoxide anions levels are presently under investigation. Further studies on the shape dependency of AgNPs in antimicrobial preparations could help to design an agent with the highest efficacy and low systemic toxicity [4]. This study aimed to investigate the mechanism of cytotoxic and genotoxic effects of AgNPs on human skin keratinocytes (HaCaT) cells. Primary cultures of HaCaT cells were maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with penicillin-streptomycin and supplement mix of fetal calf serum at 37 °C in a 5% CO2 atmosphere. They were then treated with in the presence and absences of 15μM of AgNPs for 2, 6 and 24 h. Genocytotoxic effects of AgNPs was assessed by the changes in various cellular parameters of HaCaT cells such as viability, superoxide anion radical production, lactate dehydrogenase release and the levels of the antioxidant enzymes, namely, catalase (CAT), glutathione peroxidase (GPX) and superoxide dismutase (SOD). Superoxide anion was detected using nitroblue tetrazolium NBT reduction assay. An LDH level was evaluated using the standard LDH kit, and activity of antioxidant enzymes such as CAT, GPX and SOD-1 were quantified using qPCR. Our results showed that AgNPs exposure can cause cellular oxidative damage accompanied by excessive production of superoxide anion and a significant reduction in antioxidant, but the low cytotoxic effect was observed. This suggested that AgNPs may have a potential toxic effect on the skin level only after long-term exposure.

Where applicable, experiments conform with Society ethical requirements