In Vitro Evaluation of DNA Damage Induction by Silver (Ag), Gold (Au), Silica (SiO₂), and Aluminum Oxide (Al₂O₃) Nanoparticles in Human Peripheral Blood Mononuclear Cells

Abstract

Nanoparticles (NPs) are increasingly applied in a wide range of technological and medical applications. While their use offers numerous benefits, it also raises concerns regarding their safety. Therefore, understanding their cytotoxic effects and DNA-damaging properties is crucial for ensuring the safe application of NPs. In this study, DNA-damaging properties of PVP-coated silver, silica, aluminum oxide (13 nm and 50 nm), and gold (5 nm and 40 nm) NPs in human peripheral blood mononuclear cells (PBMCs) were investigated. NPs‘ internalization and induction of reactive oxygen species were evaluated using flow cytometry. Cytotoxic properties were determined using a dual acridine orange/ethidium bromide staining technique while DNA-damaging properties were assessed using an alkaline comet assay. We observed that Ag, SiO2, and both sizes of Al2O3 NPs were efficiently internalized by human PBMCs, but only PVP-AgNPs (at 10–30 µg/mL) and SiO2 NPs (at concentrations > 100 µg/mL) induced significant DNA damage after a 24 h exposure. In contrast, the uptake of both sizes of gold nanoparticles was limited, though they were able to cause significant DNA damage after a 3 h exposure. These findings highlight the different responses of human PBMCs to various NPs, emphasizing the importance of their size, composition, and internalization rates in nanotoxicology testing.

Keywords

• nanoparticles
• DNA damage
• cytotoxicity
• comet assay
• reactive oxygen species