Sensitive detection of DNA oxidation damage induced by nanomaterials.
From a toxicological point of view, nanomaterials are of interest; because - on account of their great surface area relative to mass - they tend to be more reactive than the bulk chemicals from which they are derived. They might in some cases have the potential to damage DNA directly, or could act via the induction of oxidative stress. The comet assay (single cell gel electrophoresis) is widely used to measure DNA strand breaks and also oxidised bases, by including in the procedure digestion with lesion-specific enzymes such as formamidopyrimidine DNA glycosylase (which converts oxidised purines to breaks) or endonuclease III (recognising oxidised pyrimidines). We summarise reports in which these enzymes have been used to study a variety of nanomaterials in diverse cell types. We also stress that it is important to carry out tests of cell viability alongside the genotoxicity assay, since cytotoxicity can lead to adventitious DNA damage. Different concentrations of nanomaterials should be investigated, concentrating on a non-cytotoxic range; and incubating for short and longer periods can give valuable information about the mode of damage induction. The use of lesion-specific enzymes can substantially enhance the sensitivity of the comet assay in detecting genotoxic effects.
University of Oslo, Department of Nutrition, Oslo, Norway; NorGenotech AS, Skreia, Norway. Electronic address: a.r.collins@medisin.uio.no.
Andrew
A
Collins
NorGenotech AS, Skreia, Norway; Norwegian Institute for Air Research, Department of Environmental Chemistry, Kjeller, Norway.
Naouale
N
El Yamani
Norwegian Institute for Air Research, Department of Environmental Chemistry, Kjeller, Norway.
Maria
M
Dusinska
D002470:Cell Survival; D020552:Comet Assay / Q000379:methods*; D004247:DNA / Q000032:analysis*; D004249:DNA Damage; D045647:DNA Glycosylases / Q000378:metabolism*; D004260:DNA Repair; D006801:Humans; D049329:Nanostructures / Q000737:chemistry*; D010084:Oxidation-Reduction; D018384:Oxidative Stress
D004247:DNA; D045647:DNA Glycosylases
Comet assay; DNA oxidation; Lesion-specific enzymes; Nanomaterials