{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,9]],"date-time":"2026-03-09T18:31:16Z","timestamp":1773081076571,"version":"3.50.1"},"reference-count":53,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,1,5]],"date-time":"2026-01-05T00:00:00Z","timestamp":1767571200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Toxicol."],"abstract":"<jats:p>\n                    The reduction in the number of animals being used in experimental assays has been a concern of the scientific community. In this sense, non-animal alternative methods have been increasingly tested. This study intended to explore how cell-based responses compare to organismal outcomes and if the former models could contribute to minimizing the number of live animals needed in subsequent stages of hazard\/risk assessment of chemicals on amphibians. For this, the toxicity of the commonly used solvent dimethyl sulfoxide (DMSO) was assessed in early life stages (embryos and tadpoles) of two anuran species (\n                    <jats:italic>Xenopus laevis<\/jats:italic>\n                    and\n                    <jats:italic>Pelophylax perezi<\/jats:italic>\n                    ) and in 2\u00a0cell lines of\n                    <jats:italic>X. laevis<\/jats:italic>\n                    (A6 and XTC-2). In the\n                    <jats:italic>in vivo<\/jats:italic>\n                    assays, mortality, teratogenic effects, and biometric parameters were evaluated, while for\n                    <jats:italic>in vitro<\/jats:italic>\n                    assays, the assessed endpoint was viability. Overall, the obtained data suggest similar sensitivity of both species and life stages to DMSO. The 96 h-LC\n                    <jats:sub>50<\/jats:sub>\n                    estimated for embryos and tadpoles were, respectively, 2.19% and 2.56% for\n                    <jats:italic>X. laevis<\/jats:italic>\n                    and 3.19 and 3.41 for\n                    <jats:italic>P. perezi<\/jats:italic>\n                    . The solvent DMSO induced several malformations in early life stages, which may have implications for the fitness of organisms at later stages. A slightly higher sensitivity to DMSO was observed in the\n                    <jats:italic>in vivo<\/jats:italic>\n                    approaches comparatively to\n                    <jats:italic>in vitro<\/jats:italic>\n                    approach (72 h-LC\n                    <jats:sub>50<\/jats:sub>\n                    of 3.10% and 2.62% for A6 and XTC-2, respectively), though it can not be considered significantly different. As such, it is suggested that the latter approach may be considered to serve for first screenings of the ecotoxicity of organic solvents. Such a strategy of using\n                    <jats:italic>in vitro<\/jats:italic>\n                    assays as screening tools, has the potential to reduce the number of animals to be used in subsequent\n                    <jats:italic>in vivo<\/jats:italic>\n                    testing phases by providing information for the refinement of concentrations to be tested in\n                    <jats:italic>in vivo<\/jats:italic>\n                    assays, thereby supporting both reduction and replacement objectives.\n                  <\/jats:p>","DOI":"10.3389\/ftox.2025.1672301","type":"journal-article","created":{"date-parts":[[2026,1,5]],"date-time":"2026-01-05T05:31:28Z","timestamp":1767591088000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Comparative assessment of solvents toxicity using early life stages of amphibians and cell lines: a case study with dimethyl sulfoxide"],"prefix":"10.3389","volume":"7","author":[{"given":"S\u00f3nia Dias","family":"Coelho","sequence":"first","affiliation":[]},{"given":"Diana","family":"Campos","sequence":"additional","affiliation":[]},{"given":"M\u00f3nica","family":"Almeida","sequence":"additional","affiliation":[]},{"given":"Carla","family":"Quintaneiro","sequence":"additional","affiliation":[]},{"given":"Miguel","family":"Oliveira","sequence":"additional","affiliation":[]},{"given":"Isabel","family":"Lopes","sequence":"additional","affiliation":[]}],"member":"1965","published-online":{"date-parts":[[2026,1,5]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/s12302-021-00491-1","article-title":"Interspecific sensitivity of European amphibians towards two pesticides and comparison to standard test species","volume":"33","author":"Adams","year":"2021","journal-title":"Environ. Sci. Eur."},{"key":"B2","doi-asserted-by":"publisher","first-page":"376","DOI":"10.2307\/1536954","article-title":"Body size as a factor in the metamorphosis of tadpoles","volume":"61","author":"Adolph","year":"1931","journal-title":"Biol. Bull."},{"key":"B3","doi-asserted-by":"publisher","first-page":"87","DOI":"10.1080\/10807030802615543","article-title":"Behavioral disturbances: the missing link between sub-organismal and supra-organismal responses to stress? Prospects based on aquatic research","volume":"15","author":"Amiard-Triquet","year":"2009","journal-title":"Hum. Ecol. Risk Assess."},{"key":"B4","doi-asserted-by":"publisher","first-page":"471","DOI":"10.1080\/02757540.2022.2076838","article-title":"Ecotoxic effects of the vehicle solvent dimethyl sulfoxide on Raphidocelis subcapitata, Daphnia magna and Brachionus calyciflorus","volume":"38","author":"Andrade-Vieira","year":"2022","journal-title":"Chem. Ecol."},{"key":"B5","doi-asserted-by":"publisher","first-page":"2771","DOI":"10.1002\/etc.674","article-title":"Stage\u2010dependent susceptibility to copper in Rhinella arenarum embryos and larvae","volume":"30","author":"Aronzon","year":"2011","journal-title":"Environ. Toxicol."},{"key":"B6","article-title":"Standard guide for conducting the frog embryo teratogenesis assay \u2013 Xenopus (FETAX)","volume-title":"Standard guide E1439-12","year":"2012"},{"key":"B7","article-title":"Standard guide for conducting acute toxicity tests on test materials with fishes, macroinvertebrates, and amphibians","volume-title":"Standard guide E729-96","year":"2014"},{"key":"B8","doi-asserted-by":"publisher","first-page":"67","DOI":"10.1016\/j.jobaz.2014.09.005","article-title":"Biological factors controlling developmental duration, growth and metamorphosis of the larval green toad, Bufo viridis viridis","volume":"67","author":"Bekhet","year":"2014","journal-title":"J. Basic Appl. Zoology"},{"key":"B9","doi-asserted-by":"publisher","first-page":"213","DOI":"10.1111\/ede.12035","article-title":"Developmental origins of a novel gut morphology in frogs","volume":"15","author":"Bloom","year":"2013","journal-title":"Evol. Dev."},{"key":"B10","doi-asserted-by":"publisher","first-page":"e87609","DOI":"10.1371\/journal.pone.0087609","article-title":"The B-subdomain of the Xenopus laevis XFIN KRAB-AB domain is responsible for its weaker transcriptional repressor activity compared to human ZNF10\/Kox1","volume":"9","author":"Born","year":"2014","journal-title":"PLoS ONE"},{"key":"B11","doi-asserted-by":"publisher","first-page":"141707","DOI":"10.1016\/j.scitotenv.2020.141707","article-title":"A burning issue: the effect of organic ultraviolet filter exposure on the behaviour and physiology of Daphnia magna","volume":"750","author":"Boyd","year":"2021","journal-title":"Sci. Total Environ."},{"key":"B54","doi-asserted-by":"publisher","first-page":"103759","DOI":"10.1016\/j.dci.2020.103759","article-title":"Xela DS2 and Xela VS2: two novel skin epithelial-like cell lines from adult African clawed frog (Xenopus laevis) and their response to an extracellular viral dsRNA analogue","volume":"112","author":"Bui-Marinos","year":"2020","journal-title":"Dev. Comp. Immunol."},{"key":"B12","doi-asserted-by":"publisher","first-page":"e70121","DOI":"10.1371\/journal.pone.0070121","article-title":"Larger body size at metamorphosis enhances survival, growth and performance of young cane toads (Rhinella marina)","volume":"8","author":"Cabrera-Guzm\u00e1n","year":"2013","journal-title":"PLoS One"},{"key":"B13","doi-asserted-by":"publisher","first-page":"1789","DOI":"10.3390\/molecules22111789","article-title":"Dimethyl sulfoxide (DMSO) decreases cell proliferation and TNF-\u03b1, IFN-\u03b3, and IL-2 cytokines production in cultures of peripheral blood lymphocytes","volume":"22","author":"Costa","year":"2017","journal-title":"Molecules"},{"key":"B14","doi-asserted-by":"publisher","first-page":"237","DOI":"10.1002\/jat.2550070403","article-title":"Development of a reconstituted water medium and preliminary validation of the frog embryo teratogenesis assay \u2014 Xenopus (FETAX)","volume":"7","author":"Dawson","year":"1987","journal-title":"J. Appl. Toxicol."},{"key":"B15","doi-asserted-by":"publisher","first-page":"1413","DOI":"10.1007\/s10646-022-02598-4","article-title":"Acute toxicity characterization of organic UV-filters and chronic exposure revealing multigenerational effects in Daphnia magna","volume":"31","author":"de Paula","year":"2022","journal-title":"Ecotoxicology"},{"key":"B16","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1002\/jat.2550120111","article-title":"Teratogenic assessment of four solvents using the frog embryo teratogenesis assay \u2014 Xenopus (FETAX)","volume":"12","author":"Dresser","year":"1992","journal-title":"J. Appl. Toxicol."},{"key":"B17","unstructured":"Chemical production and consumption statistics\n          \n          \n          2024"},{"key":"B18","doi-asserted-by":"publisher","first-page":"1556788","DOI":"10.3389\/fenvs.2025.1556788","article-title":"Assessing species bias in amphibian research on endocrine disruptors: beyond Xenopus laevis","volume":"13","author":"Fr\u0105tczak","year":"2025","journal-title":"Front. Environ. Sci."},{"key":"B19","doi-asserted-by":"publisher","first-page":"4472","DOI":"10.3390\/molecules27144472","article-title":"Dimethyl sulfoxide: a bio-friendly or bio-hazard chemical? The effect of DMSO in human fibroblast-like synoviocytes","volume":"27","author":"Gallardo-Villagr\u00e1n","year":"2022","journal-title":"Molecules"},{"key":"B20","doi-asserted-by":"publisher","first-page":"1317","DOI":"10.1096\/fj.13-235440","article-title":"Unexpected low\u2010dose toxicity of the universal solvent DMSO","volume":"28","author":"Galvao","year":"2014","journal-title":"FASEB J."},{"key":"B55","first-page":"183","article-title":"Simplified table for staging anuran embryos and larvae with notes on identification","volume":"16","author":"Gosner","year":"1960","journal-title":"Herpetologica"},{"key":"B22","doi-asserted-by":"publisher","first-page":"804033","DOI":"10.3389\/ftox.2021.804033","article-title":"DMSO concentrations up to 1% are safe to be used in the zebrafish embryo developmental toxicity assay","volume":"3","author":"Hoyberghs","year":"2021","journal-title":"Front. Toxicol."},{"key":"B56","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1111\/j.1749-7345.1992.tb00766.x","article-title":"Reporting fish growth: a review on the basics","volume":"23","author":"Hopkins","year":"1992","journal-title":"J. World Aquac. Soc."},{"key":"B23","doi-asserted-by":"publisher","first-page":"107","DOI":"10.1016\/j.scitotenv.2017.09.260","article-title":"Unsuitable use of DMSO for assessing behavioral endpoints in aquatic model species","volume":"615","author":"Huang","year":"2018","journal-title":"Sci. Total Environ."},{"key":"B24","volume-title":"Global assessment report on biodiversity and ecosystem services of the intergovernmental science-policy platform on biodiversity and ecosystem services (vers\u00e3o 1)","author":"Brondizio","year":"2019"},{"key":"B25","unstructured":"The IUCN Red List of threatened species. IUCN Red List of Threatened Species\n          \n          \n          2023"},{"key":"B26","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/B978-0-323-85215-9.00047-7","article-title":"Alternative methods to animal experimentation and their role in modern toxicology","volume-title":"Toxicological risk assessment and multi-system health impacts from exposure","author":"Kandarova","year":"2021"},{"key":"B27","doi-asserted-by":"publisher","first-page":"129405","DOI":"10.1016\/j.chemosphere.2020.129405","article-title":"Combined toxicity of dimethyl sulfoxide (DMSO) and vanadium towards zebrafish embryos (Danio rerio): unexpected synergistic effect by DMSO","volume":"270","author":"Kim","year":"2021","journal-title":"Chemosphere"},{"key":"B29","doi-asserted-by":"publisher","first-page":"347","DOI":"10.1038\/s41684-024-01476-2","article-title":"Toward a common interpretation of the 3Rs principles in animal research","volume":"53","author":"Lauwereyns","year":"2024","journal-title":"LabAnimal"},{"key":"B30","doi-asserted-by":"publisher","first-page":"211","DOI":"10.1007\/s00204-008-0279-9","article-title":"Alternative methods to safety studies in experimental animals: role in the risk assessment of chemicals under the new European Chemicals Legislation (REACH)","volume":"82","author":"Lilienblum","year":"2008","journal-title":"Archives Toxicol."},{"key":"B31","doi-asserted-by":"publisher","first-page":"889","DOI":"10.1016\/j.chemosphere.2005.07.063","article-title":"Solvent toxicity to amphibian embryos and larvae","volume":"63","author":"Marquis","year":"2006","journal-title":"Chemosphere"},{"key":"B32","doi-asserted-by":"publisher","first-page":"29120241536","DOI":"10.1098\/rspb.2024.1536","article-title":"The impact of chemical pollution across major life transitions: a meta-analysis on oxidative stress in amphibians","volume":"291","author":"Martin","year":"2024","journal-title":"Proc. R. Soc. B"},{"key":"B33","doi-asserted-by":"publisher","first-page":"358","DOI":"10.1242\/jeb.151043","article-title":"Reversible developmental stasis in response to nutrient availability in the Xenopus laevis central nervous system","volume":"220","author":"McKeown","year":"2017","journal-title":"J. Exp. Biol."},{"key":"B34","doi-asserted-by":"publisher","first-page":"e200908","DOI":"10.2903\/j.efsa.2022.e200908","article-title":"The use of NAMs and omics data in risk assessment","volume":"20","author":"Miccoli","year":"2022","journal-title":"EFSA J."},{"key":"B35","doi-asserted-by":"publisher","first-page":"932","DOI":"10.1186\/s12967-025-06807-6","article-title":"Dimethyl sulfoxide in cryopreserved mesenchymal stromal cell therapy products: is there a safety risk to patients?","volume":"23","author":"Niebergall-Roth","year":"2025","journal-title":"J. Transl. Med."},{"key":"B36","volume-title":"Normal table of","author":"Nieuwkoop","year":"1994"},{"key":"B37","volume-title":"Guidance document for the diagnosis of endocrine-related histopathology of fish gonads","author":"Development","year":"2009"},{"key":"B38","volume-title":"Test No. 203 - fish acute toxicity test","author":"Development","year":"2019"},{"key":"B39","volume-title":"Test No. 249 - fish Cell Line Acute Toxicity - the RTgill-W1 cell line assay","author":"Development","year":"2021"},{"key":"B40","first-page":"31","article-title":"Effect of quercetin and DMSO on skeletal myogenesis from C2C12 skeletal muscle cells with special reference to PKB\/Akt activity, myogenin and Bcl-2 expression","volume":"11","author":"Orzechowski","year":"2001","journal-title":"Basic Appl. Myology"},{"key":"B41","article-title":"Cell viability assays","volume-title":"Assay guidance manual","author":"Ris","year":"2016"},{"key":"B43","doi-asserted-by":"publisher","first-page":"1864","DOI":"10.1002\/etc.2257","article-title":"Salinity and copper interactive effects on perez\u2019s frog Pelophylax perezi","volume":"32","author":"Santos","year":"2013","journal-title":"Environ. Toxicol. Chem."},{"key":"B44","doi-asserted-by":"publisher","first-page":"158746","DOI":"10.1016\/j.scitotenv.2022.158746","article-title":"Effects of aqueous extracts of wildfire ashes on tadpoles of Pelophylax perezi: influence of plant coverage","volume":"854","author":"Santos","year":"","journal-title":"S"},{"key":"B45","doi-asserted-by":"publisher","first-page":"106664","DOI":"10.1016\/j.aquatox.2023.106664","article-title":"Impacts of aqueous extracts of wildfire ashes on aquatic life-stages of Xenopus laevis: influence of plant coverage","volume":"262","author":"Santos","year":"","journal-title":"Aquat. Toxicol."},{"key":"B46","doi-asserted-by":"publisher","first-page":"29","DOI":"10.3390\/toxics8020029","article-title":"Effect of 10 UV filters on the brine shrimp Artemia salina and the marine microalga Tetraselmis sp","volume":"8","author":"Thorel","year":"2020","journal-title":"Toxics"},{"key":"B47","doi-asserted-by":"publisher","first-page":"830","DOI":"10.1080\/14786419.2018.1508147","article-title":"Allurin and egg jelly coat impact on in-vitro fertilization success of endangered Albanian water frog, Pelophylax shqipericus","volume":"34","author":"Turani","year":"2020","journal-title":"Nat. Prod. Res."},{"key":"B48","doi-asserted-by":"publisher","first-page":"593","DOI":"10.1002\/etc.1718","article-title":"Implications of the solvent vehicles dimethylformamide and dimethylsulfoxide for establishing transcriptomic endpoints in the zebrafish embryo toxicity test","volume":"31","author":"Turner","year":"2012","journal-title":"Environ. Toxicol."},{"key":"B49","doi-asserted-by":"publisher","first-page":"4641","DOI":"10.1038\/s41598-019-40660-0","article-title":"DMSO induces drastic changes in human cellular processes and epigenetic landscape in vitro","volume":"9","author":"Verheijen","year":"2019","journal-title":"Sci. Rep."},{"key":"B50","doi-asserted-by":"publisher","first-page":"209","DOI":"10.1641\/0006-3568(2001)051[0209:EOCOBB]2.0.CO;2","article-title":"Effects of contaminants on behavior: biochemical mechanisms and ecological consequences: killifish from a 830 contaminated site are slow to capture prey and escape predators; altered 831 neurotransmitters and thyroid may be responsible for this behavior, which may 832 produce population changes in the fish and their major prey, the grass shrimp","volume":"51","author":"Weis","year":"2001","journal-title":"Bioscience"},{"key":"B51","doi-asserted-by":"publisher","first-page":"483","DOI":"10.1002\/jat.3920","article-title":"Assessment of sublethal ecotoxicity of solvents on larvae of a model native amphibian (Lithobates pipiens)","volume":"40","author":"Young","year":"2020","journal-title":"J. Appl. Toxicol."},{"key":"B52","doi-asserted-by":"publisher","first-page":"e107447","DOI":"10.1371\/journal.pone.0107447","article-title":"Dimethyl sulfoxide damages mitochondrial integrity and membrane potential in cultured astrocytes","volume":"9","author":"Yuan","year":"2014","journal-title":"PLoS One"},{"key":"B53","doi-asserted-by":"publisher","first-page":"153","DOI":"10.1016\/S0925-4773(01)00341-0","article-title":"Gene expression in the embryonic xenopus liver","volume":"103","author":"Zorn","year":"2001","journal-title":"Mech. Dev."}],"container-title":["Frontiers in Toxicology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/ftox.2025.1672301\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,5]],"date-time":"2026-01-05T05:31:31Z","timestamp":1767591091000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/ftox.2025.1672301\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1,5]]},"references-count":53,"alternative-id":["10.3389\/ftox.2025.1672301"],"URL":"https:\/\/doi.org\/10.3389\/ftox.2025.1672301","relation":{},"ISSN":["2673-3080"],"issn-type":[{"value":"2673-3080","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,5]]},"article-number":"1672301"}}