Suitability of Salivary Leucocytes to Assess DNA Repair Ability in Human Biomonitoring Studies by the Challenge-Comet Assay
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http://hdl.handle.net/2183/31866
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Suitability of Salivary Leucocytes to Assess DNA Repair Ability in Human Biomonitoring Studies by the Challenge-Comet AssayAutor(es)
Data
2022Cita bibliográfica
Natalia Fernández-Bertólez, Carlota Lema-Arranz, Sónia Fraga, João Paulo Teixeira, Eduardo Pásaro, Laura Lorenzo-López, Vanessa Valdiglesias, Blanca Laffon (2022). Suitability of salivary leucocytes to assess DNA repair ability in human biomonitoring studies by the challenge-comet assay. Chemosphere, Volume 307, Part 4, 136139. ISSN 0045-6535. https://doi.org/10.1016/j.chemosphere.2022.136139. (https://www.sciencedirect.com/science/article/pii/S0045653522026327)
Resumo
[Abstract] The challenge-comet assay is a simple but effective approach that provides a quantitative and functional determination of DNA repair ability, and allows to monitor the kinetics of repair process. Peripheral blood mononuclear cells (PBMC) are the cells most frequently employed in human biomonitoring studies using the challenge-comet assay, but having a validated alternative of non-invasive biomatrix would be highly convenient for certain population groups and circumstances. The objective of this study was to validate the use of salivary leucocytes in the challenge-comet assay. Leucocytes were isolated from saliva samples and challenged (either in fresh or after cryopreservation) with three genotoxic agents acting by different action mechanisms: bleomycin, methyl methanesulfonate, and ultraviolet radiation. Comet assay was performed just after treatment and at other three additional time points, in order to study repair kinetics. The results obtained demonstrated that saliva leucocytes were as suitable as PBMC for assessing DNA damage of different nature that was efficiently repaired over the evaluated time points, even after 5 months of cryopreservation (after a 24 h stimulation with PHA). Furthermore, a new parameter to determine the efficacy of the repair process, independent of the initial amount of damage induced, is proposed, and recommendations to perform the challenge-comet assay with salivary leucocytes depending on the type of DNA repair to be assessed are suggested. Validation studies are needed to verify whether the method is reproducible and results reliable and comparable among laboratories and studies.
Palabras chave
Challenge-Comet assay
DNA damage Response
Frozen samples
Human biomonitoring
Salivary leucocytes
DNA damage Response
Frozen samples
Human biomonitoring
Salivary leucocytes
Descrición
Financiado para publicación en acceso aberto: Universidade da Coruña/CISUG
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Dereitos
Atribución-NoComercial-SinDerivadas 4.0 Internacional (CC BY-NC-ND 4.0)
ISSN
0045-6535