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Suitability of salivary leucocytes to assess DNA repair ability in human biomonitoring studies by the challenge-comet assay

2022-11Journal article Restricted access
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dc.contributor.authorFernández-Bertólez, Natalia
dc.contributor.authorLema-Arranz, Carlota
dc.contributor.authorFraga, Sónia
dc.contributor.authorTeixeira, João Paulo
dc.contributor.authorPásaro, Eduardo
dc.contributor.authorLorenzo-López, Laura
dc.contributor.authorValdiglesias, Vanessa
dc.contributor.authorLaffon, Blanca
dc.date.accessioned2023-03-22T11:10:15Z
dc.date.available2023-03-22T11:10:15Z
dc.date.issued2022-11
dc.description.abstractThe 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.pt_PT
dc.description.abstractHighlights: Objective: to validate the use of salivary leucocytes in the challenge-comet assay; DNA damage of different nature was repaired over the evaluated time points; Results demonstrated that salivary leucocytes were as suitable as PBMC for this assay; Results were similar in fresh and after 5 months of cryopreservation; Recommendations are given depending on the type of DNA repair to be assessed.pt_PT
dc.description.sponsorshipThis work was funded by the Spanish Ministry of Science and Innovation: MCIN/AEI/10.13039/501100011033 (Grants PID2020- 113788RB-I00 and PID2020-114908 GA-I00), NanoBioBarriers project (PTDC/MED-TOX/31162/2017), Xunta de Galicia (ED431B 2022/16), co-financed by the Operational Program for Competitiveness and Internationalization (POCI) through European Regional Development Funds (FEDER/FNR), Spanish Ministry of Education, Culture and Sport [BEAGAL18/00142 to V.V.], and Spanish Ministry of Economy and Competitiveness, co-financed by the European Social Fund [RYC-2015- 18394 to L.L,-L,]. Fundingpt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationChemosphere. 2022 Nov;307(Pt 4):136139. doi: 10.1016/j.chemosphere.2022.136139. Epub 2022 Aug 22pt_PT
dc.identifier.doi10.1016/j.chemosphere.2022.136139pt_PT
dc.identifier.issn0045-6535
dc.identifier.urihttp://hdl.handle.net/10400.18/8597
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherElsevierpt_PT
dc.relation.publisherversionhttps://www.sciencedirect.com/science/article/pii/S0045653522026327?via%3Dihubpt_PT
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/pt_PT
dc.subjectChallenge-Comet Assaypt_PT
dc.subjectDNA Damage Responsept_PT
dc.subjectFrozen Samplespt_PT
dc.subjectHuman Biomonitoringpt_PT
dc.subjectSalivary Leucocytespt_PT
dc.subjectGenotoxicidade Ambientalpt_PT
dc.titleSuitability of salivary leucocytes to assess DNA repair ability in human biomonitoring studies by the challenge-comet assaypt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/9471 - RIDTI/PTDC%2FMED-TOX%2F31162%2F2017/PT
oaire.citation.issuePt 4pt_PT
oaire.citation.startPage136139pt_PT
oaire.citation.titleChemospherept_PT
oaire.citation.volume307pt_PT
oaire.fundingStream9471 - RIDTI
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.embargofctAcesso de acordo com política editorial da revista.pt_PT
rcaap.rightsembargoedAccesspt_PT
rcaap.typearticlept_PT
relation.isProjectOfPublication1370de00-d228-47a7-8aed-320831dc4dcd
relation.isProjectOfPublication.latestForDiscovery1370de00-d228-47a7-8aed-320831dc4dcd

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