Development of an assay to assess genotoxicity by particulate matter extract

Mol Med Rep. 2017 Apr;15(4):1738-1746. doi: 10.3892/mmr.2017.6171. Epub 2017 Feb 6.

Abstract

The current study describes a method for assessing the oxidative potential of common environmental stressors (ambient air particulate matter), using a plasmid relaxation assay where the extract caused single-strand breaks, easily visualised through electrophoresis. This assay utilises a miniscule amount (11 µg) of particulate matter (PM) extract compared to other, cell‑based methods (~3,000 µg). The negative impact of air pollution on human health has been extensively recognised. Among the air pollutants, PM plays an eminent role, as reflected in the broad scientific interest. PM toxicity highly depends on its composition (metals and organic compounds), which in turn has been linked to multiple health effects (such as cardiorespiratory diseases and cancer) through multiple toxicity mechanisms; the induction of oxidative stress is considered a major mechanism among these. In this study, the PM levels, oxidative potential, cytotoxicity and genotoxicity of PM in the region of Larissa, Greece were examined using the plasmid relaxation assay. Finally, coffee extracts from different varieties, derived from both green and roasted seeds, were examined for their ability to inhibit PM-induced DNA damage. These extracts also exerted an inhibitory effect on xanthine oxidase and catalase, but had no effect against superoxide dismutase. Overall, this study highlights the importance of assays for assessing the oxidative potential of widespread environmental stressors (PM), as well as the antioxidant capacity of beverages and food items, with the highlight being the development of a plasmid relaxation assay to assess the genotoxicity caused by PM using only a miniscule amount.

MeSH terms

  • Antioxidants / pharmacology
  • Cell Death / drug effects
  • Coffea / chemistry
  • DNA Cleavage / drug effects
  • DNA Damage*
  • Humans
  • Mutagenicity Tests / methods*
  • Particulate Matter / toxicity*
  • Plant Extracts / pharmacology
  • Polyphenols / analysis

Substances

  • Antioxidants
  • Particulate Matter
  • Plant Extracts
  • Polyphenols