Protective effect of supplementation with water soluble β-carotene and α-tocopherol in boar sperm cooling-freezing extender, but not in the thawing extender

Cryopreservation of sperm is a crucial tool for the long-term preservation of male genetic material, causing significant issues in motility, membrane, and acrosome integrity, among other parameters. Antioxidants have been used to cope with these detrimental effects. We tested 1. the toxicity of a wi...

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Autores: Martín-San Juan, Adrián, de Mercado, Eduardo, Nieto-Cristóbal, Helena, Cabero, Andrea, Silvestre, Miguel Ángel, Morrell, J. M., Álvarez-Rodríguez, Manuel
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2025
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/394921
Acceso en línea:http://hdl.handle.net/10261/394921
https://api.elsevier.com/content/abstract/scopus_id/85217064891
Access Level:acceso abierto
Palabra clave:Cryopreservation
Pig
Thermal-resistance test
α-tocopherol
β-carotene
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spelling Protective effect of supplementation with water soluble β-carotene and α-tocopherol in boar sperm cooling-freezing extender, but not in the thawing extenderMartín-San Juan, Adriánde Mercado, EduardoNieto-Cristóbal, HelenaCabero, AndreaSilvestre, Miguel ÁngelMorrell, J. M.Álvarez-Rodríguez, ManuelCryopreservationPigThermal-resistance testα-tocopherolβ-caroteneCryopreservation of sperm is a crucial tool for the long-term preservation of male genetic material, causing significant issues in motility, membrane, and acrosome integrity, among other parameters. Antioxidants have been used to cope with these detrimental effects. We tested 1. the toxicity of a wide range of water soluble β-carotene (β: 250–4000 µM) and α-tocopherol (α: 31–496 µM) concentrations on boar ejaculated sperm (n = 6) in parameters as motility, viability, acrosome reaction, apoptosis, oxidation, mitochondrial activation and membrane potential; 2. the effect of various β-carotene (250–1000 µM) and α-tocopherol (31–124 µM) concentrations added to the cooling-freezing or thawing extenders (n = 30) before (0 min) and after 90 min incubation (37 °C). Toxicity results showed a decrease in the proportion of live spermatozoa with non-reacted acrosome from 75.1 ± 3.3 % using β250/α31 to 60.1 ± 5.7 % and 59.3 ± 5.4 % in samples with β2000/α248 and β4000/α496 respectively (p < 0.05), suggesting a detrimental effect of the highest concentrations. Antioxidant supplementation in the cooling-freezing extender decreased the apoptotic and oxidized spermatozoa in β500/α62 and β1000/α124, relative to the control. In contrast, antioxidants addition to the thawing extender induced some detrimental effects in several sperm parameters analyzed. In conclusion, water-soluble β-carotene and α-tocopherol prevent acrosome reaction and oxidation during cooling-freezing on boar sperm. High concentrations of these antioxidants negatively impacted motility and mitochondrial function, suggesting cytotoxic effects and potential capacitation-like changes. The β1000/α124 showed protective effects during cryopreservation, but post-thawing supplementation may stimulate oxidative stress rather than prevent it.This research was funded by the projects PID2019-108320RJI00, RYC2020-028615-I, PID2022-136561OB-I00, CNS2023-144564 and CIAICO/2022/174 funded by MCIN/AEI/10.13039/501100011033 (Spain), Generalitat Valenciana and FEDER, FSE and NextGenerationEU/PRTR Funds (EU). A.C. developed part of the experimental approaches during an internship as a Final Degree Project (Universidad de Alcalá, Madrid, Spain).Peer reviewedElsevier BVAgencia Estatal de Investigación (España)Ministerio de Ciencia, Innovación y Universidades (España)Generalitat ValencianaEuropean CommissionUniversidad de AlcaláCabero, Andrea [0009-0008-7978-2143]Silvestre, Miguel Ángel [0000-0001-9260-5792]Morrell, J. M. [0000-0002-5245-7331]Álvarez-Rodríguez, Manuel 0000-0003-0120-354XConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202520252025info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/394921https://api.elsevier.com/content/abstract/scopus_id/85217064891reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/PID2022-136561OB-I00info:eu-repo/grantAgreement/AEI//CNS2023-144564info:eu-repo/grantAgreement/AEI//CIAICOThe underlying dataset has been published as supplementary material of the article in the publisher platform at DOI https://doi.org/10.1016/j.anireprosci.2025.107792https://doi.org/10.1016/j.anireprosci.2025.107792Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/3949212026-05-22T06:33:51Z
dc.title.none.fl_str_mv Protective effect of supplementation with water soluble β-carotene and α-tocopherol in boar sperm cooling-freezing extender, but not in the thawing extender
title Protective effect of supplementation with water soluble β-carotene and α-tocopherol in boar sperm cooling-freezing extender, but not in the thawing extender
spellingShingle Protective effect of supplementation with water soluble β-carotene and α-tocopherol in boar sperm cooling-freezing extender, but not in the thawing extender
Martín-San Juan, Adrián
Cryopreservation
Pig
Thermal-resistance test
α-tocopherol
β-carotene
title_short Protective effect of supplementation with water soluble β-carotene and α-tocopherol in boar sperm cooling-freezing extender, but not in the thawing extender
title_full Protective effect of supplementation with water soluble β-carotene and α-tocopherol in boar sperm cooling-freezing extender, but not in the thawing extender
title_fullStr Protective effect of supplementation with water soluble β-carotene and α-tocopherol in boar sperm cooling-freezing extender, but not in the thawing extender
title_full_unstemmed Protective effect of supplementation with water soluble β-carotene and α-tocopherol in boar sperm cooling-freezing extender, but not in the thawing extender
title_sort Protective effect of supplementation with water soluble β-carotene and α-tocopherol in boar sperm cooling-freezing extender, but not in the thawing extender
dc.creator.none.fl_str_mv Martín-San Juan, Adrián
de Mercado, Eduardo
Nieto-Cristóbal, Helena
Cabero, Andrea
Silvestre, Miguel Ángel
Morrell, J. M.
Álvarez-Rodríguez, Manuel
author Martín-San Juan, Adrián
author_facet Martín-San Juan, Adrián
de Mercado, Eduardo
Nieto-Cristóbal, Helena
Cabero, Andrea
Silvestre, Miguel Ángel
Morrell, J. M.
Álvarez-Rodríguez, Manuel
author_role author
author2 de Mercado, Eduardo
Nieto-Cristóbal, Helena
Cabero, Andrea
Silvestre, Miguel Ángel
Morrell, J. M.
Álvarez-Rodríguez, Manuel
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Agencia Estatal de Investigación (España)
Ministerio de Ciencia, Innovación y Universidades (España)
Generalitat Valenciana
European Commission
Universidad de Alcalá
Cabero, Andrea [0009-0008-7978-2143]
Silvestre, Miguel Ángel [0000-0001-9260-5792]
Morrell, J. M. [0000-0002-5245-7331]
Álvarez-Rodríguez, Manuel 0000-0003-0120-354X
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Cryopreservation
Pig
Thermal-resistance test
α-tocopherol
β-carotene
topic Cryopreservation
Pig
Thermal-resistance test
α-tocopherol
β-carotene
description Cryopreservation of sperm is a crucial tool for the long-term preservation of male genetic material, causing significant issues in motility, membrane, and acrosome integrity, among other parameters. Antioxidants have been used to cope with these detrimental effects. We tested 1. the toxicity of a wide range of water soluble β-carotene (β: 250–4000 µM) and α-tocopherol (α: 31–496 µM) concentrations on boar ejaculated sperm (n = 6) in parameters as motility, viability, acrosome reaction, apoptosis, oxidation, mitochondrial activation and membrane potential; 2. the effect of various β-carotene (250–1000 µM) and α-tocopherol (31–124 µM) concentrations added to the cooling-freezing or thawing extenders (n = 30) before (0 min) and after 90 min incubation (37 °C). Toxicity results showed a decrease in the proportion of live spermatozoa with non-reacted acrosome from 75.1 ± 3.3 % using β250/α31 to 60.1 ± 5.7 % and 59.3 ± 5.4 % in samples with β2000/α248 and β4000/α496 respectively (p < 0.05), suggesting a detrimental effect of the highest concentrations. Antioxidant supplementation in the cooling-freezing extender decreased the apoptotic and oxidized spermatozoa in β500/α62 and β1000/α124, relative to the control. In contrast, antioxidants addition to the thawing extender induced some detrimental effects in several sperm parameters analyzed. In conclusion, water-soluble β-carotene and α-tocopherol prevent acrosome reaction and oxidation during cooling-freezing on boar sperm. High concentrations of these antioxidants negatively impacted motility and mitochondrial function, suggesting cytotoxic effects and potential capacitation-like changes. The β1000/α124 showed protective effects during cryopreservation, but post-thawing supplementation may stimulate oxidative stress rather than prevent it.
publishDate 2025
dc.date.none.fl_str_mv 2025
2025
2025
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/394921
https://api.elsevier.com/content/abstract/scopus_id/85217064891
url http://hdl.handle.net/10261/394921
https://api.elsevier.com/content/abstract/scopus_id/85217064891
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
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info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/PID2022-136561OB-I00
info:eu-repo/grantAgreement/AEI//CNS2023-144564
info:eu-repo/grantAgreement/AEI//CIAICO
The underlying dataset has been published as supplementary material of the article in the publisher platform at DOI https://doi.org/10.1016/j.anireprosci.2025.107792
https://doi.org/10.1016/j.anireprosci.2025.107792
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
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dc.publisher.none.fl_str_mv Elsevier BV
publisher.none.fl_str_mv Elsevier BV
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
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