Suitability of salivary leucocytes to assess DNA repair ability in human biomonitoring studies by the challenge-comet assay
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.18/8597 |
Resumo: | 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. |
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Suitability of salivary leucocytes to assess DNA repair ability in human biomonitoring studies by the challenge-comet assayChallenge-Comet AssayDNA Damage ResponseFrozen SamplesHuman BiomonitoringSalivary LeucocytesGenotoxicidade AmbientalThe 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.Highlights: 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.This 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,]. FundingElsevierRepositório Científico do Instituto Nacional de SaúdeFernández-Bertólez, NataliaLema-Arranz, CarlotaFraga, SóniaTeixeira, João PauloPásaro, EduardoLorenzo-López, LauraValdiglesias, VanessaLaffon, Blanca2023-03-22T11:10:15Z2022-112022-11-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.18/8597engChemosphere. 2022 Nov;307(Pt 4):136139. doi: 10.1016/j.chemosphere.2022.136139. Epub 2022 Aug 220045-653510.1016/j.chemosphere.2022.136139info:eu-repo/semantics/embargoedAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-20T15:42:39Zoai:repositorio.insa.pt:10400.18/8597Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T18:43:14.699963Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Suitability of salivary leucocytes to assess DNA repair ability in human biomonitoring studies by the challenge-comet assay |
title |
Suitability of salivary leucocytes to assess DNA repair ability in human biomonitoring studies by the challenge-comet assay |
spellingShingle |
Suitability of salivary leucocytes to assess DNA repair ability in human biomonitoring studies by the challenge-comet assay Fernández-Bertólez, Natalia Challenge-Comet Assay DNA Damage Response Frozen Samples Human Biomonitoring Salivary Leucocytes Genotoxicidade Ambiental |
title_short |
Suitability of salivary leucocytes to assess DNA repair ability in human biomonitoring studies by the challenge-comet assay |
title_full |
Suitability of salivary leucocytes to assess DNA repair ability in human biomonitoring studies by the challenge-comet assay |
title_fullStr |
Suitability of salivary leucocytes to assess DNA repair ability in human biomonitoring studies by the challenge-comet assay |
title_full_unstemmed |
Suitability of salivary leucocytes to assess DNA repair ability in human biomonitoring studies by the challenge-comet assay |
title_sort |
Suitability of salivary leucocytes to assess DNA repair ability in human biomonitoring studies by the challenge-comet assay |
author |
Fernández-Bertólez, Natalia |
author_facet |
Fernández-Bertólez, Natalia Lema-Arranz, Carlota Fraga, Sónia Teixeira, João Paulo Pásaro, Eduardo Lorenzo-López, Laura Valdiglesias, Vanessa Laffon, Blanca |
author_role |
author |
author2 |
Lema-Arranz, Carlota Fraga, Sónia Teixeira, João Paulo Pásaro, Eduardo Lorenzo-López, Laura Valdiglesias, Vanessa Laffon, Blanca |
author2_role |
author author author author author author author |
dc.contributor.none.fl_str_mv |
Repositório Científico do Instituto Nacional de Saúde |
dc.contributor.author.fl_str_mv |
Fernández-Bertólez, Natalia Lema-Arranz, Carlota Fraga, Sónia Teixeira, João Paulo Pásaro, Eduardo Lorenzo-López, Laura Valdiglesias, Vanessa Laffon, Blanca |
dc.subject.por.fl_str_mv |
Challenge-Comet Assay DNA Damage Response Frozen Samples Human Biomonitoring Salivary Leucocytes Genotoxicidade Ambiental |
topic |
Challenge-Comet Assay DNA Damage Response Frozen Samples Human Biomonitoring Salivary Leucocytes Genotoxicidade Ambiental |
description |
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. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-11 2022-11-01T00:00:00Z 2023-03-22T11:10:15Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10400.18/8597 |
url |
http://hdl.handle.net/10400.18/8597 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Chemosphere. 2022 Nov;307(Pt 4):136139. doi: 10.1016/j.chemosphere.2022.136139. Epub 2022 Aug 22 0045-6535 10.1016/j.chemosphere.2022.136139 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/embargoedAccess |
eu_rights_str_mv |
embargoedAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Elsevier |
publisher.none.fl_str_mv |
Elsevier |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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