By detecting biological effects before the development of clinical diseases, the use of biomarkers of early effects in human biomonitoring allows a prompt detection of the biological consequences of a specific situation, such as the exposure to air pollutants. Furthermore, biomarkers of early effects are measurable in a wide number of individuals, representing in more appropriately the global burden of effects that environmental exposures may determine in a population. Primary DNA damage detected by the comet assay showed to be sensitive and effective in the evaluation of the impact of human exposure to environmental mutagens and carcinogens. Several studies found a positive association between high level of air pollutants (PM and ozone, in particular) and DNA damage detected by comet assay. Most of these studies used blood cells. In the MAPEC_LIFE project, we assessed primary DNA damage performing comet assay on salivary leukocytes of children exposed to different levels of air pollution. This type of cells, directly exposed to air passage through the mouth, represents the target cells of this exposure and so is more appropriate than blood cells for the evaluation of its effects. Moreover, they are easy to retrieve, especially in children, using a simple and non-invasive method, which allows to obtain a great sample size. Despite these considerations, we encountered some difficulties working with buccal cells, especially regarding the number of cells retrieved per sample, the interference played by the buccal epithelial cells on the comet images, and the consequent extremely time-consuming analysis of the slides. Nevertheless, final results showed significantly different levels of DNA damage in child cells by the sampling season and town of residence. Moreover, statistical significant associations were found between primary DNA damage in salivary leukocytes and levels of some air pollutants, ozone in particular.
Comet assay in salivary leukocytes for the evaluation of early biological effects of air pollution exposure in children.
Bonetta Sa.;
2017-01-01
Abstract
By detecting biological effects before the development of clinical diseases, the use of biomarkers of early effects in human biomonitoring allows a prompt detection of the biological consequences of a specific situation, such as the exposure to air pollutants. Furthermore, biomarkers of early effects are measurable in a wide number of individuals, representing in more appropriately the global burden of effects that environmental exposures may determine in a population. Primary DNA damage detected by the comet assay showed to be sensitive and effective in the evaluation of the impact of human exposure to environmental mutagens and carcinogens. Several studies found a positive association between high level of air pollutants (PM and ozone, in particular) and DNA damage detected by comet assay. Most of these studies used blood cells. In the MAPEC_LIFE project, we assessed primary DNA damage performing comet assay on salivary leukocytes of children exposed to different levels of air pollution. This type of cells, directly exposed to air passage through the mouth, represents the target cells of this exposure and so is more appropriate than blood cells for the evaluation of its effects. Moreover, they are easy to retrieve, especially in children, using a simple and non-invasive method, which allows to obtain a great sample size. Despite these considerations, we encountered some difficulties working with buccal cells, especially regarding the number of cells retrieved per sample, the interference played by the buccal epithelial cells on the comet images, and the consequent extremely time-consuming analysis of the slides. Nevertheless, final results showed significantly different levels of DNA damage in child cells by the sampling season and town of residence. Moreover, statistical significant associations were found between primary DNA damage in salivary leukocytes and levels of some air pollutants, ozone in particular.
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