High-throughput screening platform for engineered nanoparticle-mediated genotoxicity using CometChip technology.

Area of Science:

• Nanotechnology
• Genotoxicity
• Biomedical Engineering

Background:

• Engineered nanoparticles (ENPs) are increasingly used in consumer products, raising concerns about potential health risks.
• The genotoxic potential of ENPs, particularly their ability to damage DNA and promote cancer, remains incompletely understood.
• Assessing ENP genotoxicity requires efficient and reliable methods to evaluate DNA damage in exposed cells.

Purpose of the Study:

• To develop and validate a high-throughput screening assay for evaluating DNA damage induced by ENPs.
• To assess the genotoxicity of five industrially relevant ENPs (SiO2, ZnO, Fe2O3, Ag, CeO2) in human and Chinese hamster cell lines.
• To compare the efficacy of the novel CometChip assay with standard comet assay methods.

Main Methods:

• Utilized the CometChip technology, a microfabricated 96-well assay for high-throughput screening of DNA damage.
• Evaluated single-stranded DNA breaks, abasic sites, and alkali-sensitive sites in cells exposed to ENPs.
• Assessed cellular viability and proliferation using MTT and CyQuant NF assays in TK6 and H9T3 cell lines.

Main Results:

• Engineered nanoparticle exposure induced dose-dependent increases in DNA damage and cytotoxicity.
• Genotoxicity varied by ENP type and cell line, with observed profiles such as ZnO>Ag>Fe2O3>CeO2>SiO2 in TK6 cells and Ag>Fe2O3>ZnO>CeO2>SiO2 in H9T3 cells.
• The CometChip platform demonstrated efficient and reliable measurement of ENP-mediated DNA damage.

Conclusions:

• The CometChip assay is an effective tool for high-throughput nanogenotoxicity assessment.
• Different ENPs exhibit distinct genotoxic profiles across various cell types.
• This assay facilitates a better understanding of the health risks associated with ENP exposure.