Evaluating nanobiomaterial-induced DNA strand breaks using the alkaline comet assay

Area of Science:

• Nanomedicine
• Materials Science
• Toxicology

Background:

• Nanobiomaterials (NBMs) offer unique properties for medical applications but face toxicity concerns.
• Genotoxicity data for NBMs is limited, yet crucial for new medical device regulations requiring long-term exposure assessment.
• Existing research extensively covers immunotoxicity and cytotoxicity, but genotoxicity remains understudied.

Purpose of the Study:

• To evaluate the genotoxic potential of five distinct NBM formulations.
• To assess the applicability of the alkaline comet assay for NBM genotoxicity screening.
• To contribute to the understanding of NBM safety under new regulatory frameworks.

Main Methods:

• An interlaboratory comparison (ILC) study was conducted within the Horizon 2020 REFINE project.
• Five NBM formulations were tested: gold nanoparticle (AuNP), IR-780 dye-loaded liposome (LipImage™815), unloaded PACA nanoparticle (PACA), cabazitaxel-loaded PACA (CBZ-PACA), and NR668 dye-loaded PACA (NR668 PACA).
• The alkaline comet assay was employed to detect DNA strand breaks induced by NBMs.

Main Results:

• The alkaline comet assay demonstrated reproducibility and repeatability across laboratories for NBM genotoxicity assessment.
• The study successfully applied the alkaline comet assay to diverse NBM formulations, including nanoparticles and liposomes.
• The findings support the assay's suitability for pre-clinical evaluation of NBMs.

Conclusions:

• The alkaline comet assay is a reliable method for assessing NBM-induced DNA damage.
• This methodology is suitable for the pre-clinical genotoxicity assessment of nanobiomaterials.
• The study provides a validated workflow for NBM genotoxicity testing, aligning with regulatory demands.