Frontiers in Toxicology (May 2022)

In vivo Mammalian Alkaline Comet Assay: Method Adapted for Genotoxicity Assessment of Nanomaterials

  • Renato Cardoso,
  • Maria Dusinska,
  • Andrew Collins,
  • Mugimane Manjanatha,
  • Stefan Pfuhler,
  • Marilyn Registre,
  • Rosalie Elespuru

DOI
https://doi.org/10.3389/ftox.2022.903896
Journal volume & issue
Vol. 4

Abstract

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The in vivo Comet assay measures the generation of DNA strand breaks under conditions in which the DNA will unwind and migrate to the anode in an electrophoresis assay, producing comet-like figures. Measurements are on single cells, which allows the sampling of a diversity of cells and tissues for DNA damaging effects. The Comet assay is the most common in vivo method for genotoxicity assessment of nanomaterials (NM). The Method outlined here includes a recommended step-by-step approach, consistent with OECD 489, taking into consideration the issues impacting assessment of NM, including choice of cells or systems, handling of NM test articles, dose determination, assay methods and data assessment. This method is designed to be used along with the accompanying “Common Considerations” paper, which discusses issues common to any genotoxicity assay using NM as a test article.

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