{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,16]],"date-time":"2026-02-16T11:21:03Z","timestamp":1771240863423,"version":"3.50.1"},"reference-count":82,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2026,2,14]],"date-time":"2026-02-14T00:00:00Z","timestamp":1771027200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"AL4AnimalS","award":["LA\/P\/0059\/2020"],"award-info":[{"award-number":["LA\/P\/0059\/2020"]}]},{"name":"CIISA","award":["UIDB\/50006\/2020"],"award-info":[{"award-number":["UIDB\/50006\/2020"]}]},{"name":"CIISA","award":["UID\/CVT\/0027\/2020"],"award-info":[{"award-number":["UID\/CVT\/0027\/2020"]}]},{"name":"EU through Project Cryostore","award":["EXPL\/BIA-BQM\/1361\/2021"],"award-info":[{"award-number":["EXPL\/BIA-BQM\/1361\/2021"]}]},{"name":"EU through Project Cryostore","award":["101120454"],"award-info":[{"award-number":["101120454"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Veterinary Sciences"],"abstract":"<jats:p>Assisted reproductive technologies (ART) use has increased over the past decades. However, reports concerning ART\u2019s low efficiency continue to emerge, citing causes related to lower embryo quality and pregnancy rates compared to their in vivo counterparts. One of the setbacks of ART is oxidative stress, which can impair embryo developmental rates. Mitochondrial redox and energetic homeostasis determine both cell survival and death, so mitochondria are a key target for therapeutic intervention strategies. In the present work, our objective was to improve the quality of viable embryos by adding new mitochondria-targeted antioxidants in the embryo culture media to reduce oxidative stress. Two naturally derived antioxidants synthesized by our team, AntiOxBEN2 and AntiOxCIN4, based on hydroxybenzoic and hydroxycinnamic scaffolds, respectively, were studied in two different experimental protocols (here called experiments). The first experiment investigated the effects of the antioxidants on embryo development to determine their optimal concentrations. The first assay of the first experiment focused on the effects of AntiOxCIN4 at concentrations of 1, 2.5, and 10 \u03bcM, while the second assay focused on the effects of AntiOxBEN2 at the same concentrations. A control group without supplementation was run simultaneously. The second experiment aimed to compare the best concentrations of these antioxidant molecules in the embryo culture media and their effect on embryos\u2019 resistance to vitrification\/warming. In each experiment, the embryos were morphologically evaluated, and the total and viable cell numbers were examined. Reactive oxygen species (ROS) and mitochondrial polarization were also evaluated using specific fluorescent dyes. In experiment 1, an increased embryo quality was identified by using 2.5 \u03bcM AntiOxCIN4 (p = 0.03) and 2.5 \u03bcM AntiOxBEN2 (p = 0.001). Moreover, blastocysts supplemented with 2.5 \u03bcM AntiOxCIN4 had higher viability (p = 0.008), while those supplemented with 2.5 \u03bcM AntiOxBEN2 presented a greater total cell number (p = 0.01). An improvement in embryo cryosurvival following the supplementation during the culture process with either antioxidant was identified in experiment 2, with superior expansion scores after vitrification\/warming and culture (2.5 \u03bcM AntiOxCIN4,\u00a0p = 0.056 and 2.5 \u03bcM AntiOxBEN2,\u00a0p = 0.059). In conclusion, both AntiOxCIN4 and AntiOxBEN2 had a beneficial effect on embryo development and cryosurvival, suggesting a potential intervention to reduce oxidative stress in assisted reproductive technologies.<\/jats:p>","DOI":"10.3390\/vetsci13020190","type":"journal-article","created":{"date-parts":[[2026,2,16]],"date-time":"2026-02-16T10:14:32Z","timestamp":1771236872000},"page":"190","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Hydroxycinnamic and Hydroxybenzoic-Based Mitochondriotropic Antioxidants Improve Bovine Embryo Quality and Cryo-Survival"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-5385-4012","authenticated-orcid":false,"given":"Filipa","family":"Ferreira","sequence":"first","affiliation":[{"name":"INIAV-Instituto Nacional de Investiga\u00e7\u00e3o Agr\u00e1ria e Veterin\u00e1ria, Unidade de Biotecnologia e Recursos Gen\u00e9ticos, Quinta da Fonte Boa, 2005-048 Vale de Santar\u00e9m, Portugal"},{"name":"Geobiotec, Faculdade de Ci\u00eancias e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal"}]},{"given":"Beatriz","family":"Louren\u00e7o","sequence":"additional","affiliation":[{"name":"INIAV-Instituto Nacional de Investiga\u00e7\u00e3o Agr\u00e1ria e Veterin\u00e1ria, Unidade de Biotecnologia e Recursos Gen\u00e9ticos, Quinta da Fonte Boa, 2005-048 Vale de Santar\u00e9m, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0834-5698","authenticated-orcid":false,"given":"Jos\u00e9","family":"Teixeira","sequence":"additional","affiliation":[{"name":"CNC UC-Center for Neuroscience and Cell Biology, University of Coimbra, 3000-548 Coimbra, Portugal"},{"name":"CIBB-Center for Innovative Biomedicine and Biotechnology, University of Coimbra, 3000-548 Coimbra, Portugal"},{"name":"Department of Life Sciences, University of Coimbra, 3000-548 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9410-6874","authenticated-orcid":false,"given":"Fernando","family":"Cagide","sequence":"additional","affiliation":[{"name":"MedInUP, Center for Drug Discovery and Innovative Medicines, Department of Biomedicine, Faculty of Medicine, University of Porto, 4200-319 Porto, Portugal"},{"name":"Department of Chemistry and Biochemistry, Faculty of Sciences, University of Porto, 4169-007 Porto, Portugal"}]},{"given":"Sofia","family":"Benfeito","sequence":"additional","affiliation":[{"name":"MedInUP, Center for Drug Discovery and Innovative Medicines, Department of Biomedicine, Faculty of Medicine, University of Porto, 4200-319 Porto, Portugal"},{"name":"Department of Chemistry and Biochemistry, Faculty of Sciences, University of Porto, 4169-007 Porto, Portugal"}]},{"given":"Fernando","family":"Lidon","sequence":"additional","affiliation":[{"name":"Geobiotec, Faculdade de Ci\u00eancias e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1050-2402","authenticated-orcid":false,"given":"Fernanda","family":"Borges","sequence":"additional","affiliation":[{"name":"MedInUP, Center for Drug Discovery and Innovative Medicines, Department of Biomedicine, Faculty of Medicine, University of Porto, 4200-319 Porto, Portugal"},{"name":"Department of Chemistry and Biochemistry, Faculty of Sciences, University of Porto, 4169-007 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5201-9948","authenticated-orcid":false,"given":"Paulo J.","family":"Oliveira","sequence":"additional","affiliation":[{"name":"CNC UC-Center for Neuroscience and Cell Biology, University of Coimbra, 3000-548 Coimbra, Portugal"},{"name":"CIBB-Center for Innovative Biomedicine and Biotechnology, University of Coimbra, 3000-548 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7514-6130","authenticated-orcid":false,"given":"Rosa M. Lino Neto","family":"Pereira","sequence":"additional","affiliation":[{"name":"INIAV-Instituto Nacional de Investiga\u00e7\u00e3o Agr\u00e1ria e Veterin\u00e1ria, Unidade de Biotecnologia e Recursos Gen\u00e9ticos, Quinta da Fonte Boa, 2005-048 Vale de Santar\u00e9m, Portugal"},{"name":"CIISA-Centro de Investiga\u00e7\u00e3o Interdisciplinar em Sanidade Animal, Faculdade de Medicina Veterin\u00e1ria, Universidade de Lisboa, Av. Universidade T\u00e9cnica, 1300-477 Lisboa, Portugal"},{"name":"AL4AnimalS-Associate Laboratory for Animal and Veterinary Sciences, Faculdade de Medicina Veterin\u00e1ria, Universidade de Lisboa, Av. Universidade T\u00e9cnica, 1300-477 Lisboa, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Pereira, R., Marques, C.C., Pimenta, J., Barbas, J.P., Baptista, M.C., Diniz, P., Torres, A., and Lopes-da-Costa, L. (2020). 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