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This study investigated the toxicity of ZnO-NP and ZnO-BK using alternative in vitro and in vivo models. The Salmonella\/Microsome assay (\u00b1metabolic activation) was conducted according to the Organisation for Economic Co-operation and Development (OECD) guidelines. HepG2 cell viability and genotoxicity were assessed using the WST-1 and cytokinesis-block micronucleus (CBMN) assays. Acute toxicity was evaluated using the fish embryotoxicity (FET) test in zebrafish (Danio rerio) embryos. Both forms induced mutagenic responses in TA98 and TA102 strains, while ZnO-BK showed mutagenicity in TA100 only in the absence of metabolic activation. In HepG2 cells, both forms induced dose-dependent cytotoxicity in the WST-1 model, with ZnO-BK exhibiting greater cytotoxic potency. The CBMN assay revealed higher cytogenetic damage for ZnO-BK, including micronuclei, nuclear buds, and nucleoplasmic bridges, while both forms exhibited dose-dependent cytotoxic and cytostatic effects. In vivo, ZnO-NP exerted a more pronounced impact on embryonic development, altering eye diameter and body size compared to ZnO-BK. Overall, both forms demonstrated significant toxicological effects, highlighting their relevance for regulatory toxicogenetics and environmental safety.<\/jats:p>","DOI":"10.1021\/acsomega.6c07375","type":"journal-article","created":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T21:01:46Z","timestamp":1790110906000},"page":"60104-60118","source":"Crossref","is-referenced-by-count":0,"title":["A Comparative Analysis of Zinc Oxide as Nanoparticles\nand Its Bulk Form through In Vitro Toxicogenetics and In Vivo Zebrafish\nEmbryotoxicity Models"],"prefix":"10.1021","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9278-4441","authenticated-orcid":true,"given":"Carlos Fernando","family":"Araujo-Lima","sequence":"first","affiliation":[{"name":"Universidade Federal do Estado do Rio de Janeiro (UNIRIO) , , , ,","place":["Rio de Janeiro, RJ, Brazil, 20211-010"]},{"name":"Universidade do Estado do Rio de Janeiro (UERJ) , , , ,","place":["Rio de Janeiro, RJ, Brazil, 20551-030"]},{"name":"Universidade Federal do Estado do Rio de Janeiro (UNIRIO) , , , ,","place":["Rio de Janeiro, RJ, Brazil, 20211-010"]},{"name":"Universidade Federal do Estado do Rio de Janeiro (UNIRIO) , , , ,","place":["Rio de Janeiro, RJ, Brazil, 20211-010"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3438-1481","authenticated-orcid":true,"given":"Magno","family":"Maciel-Magalha\u0303es","sequence":"additional","affiliation":[{"name":"Fundac\u0327a\u0303o Oswaldo Cruz (FIOCRUZ) , , ,","place":["Rio de Janeiro, Brazil, 21040-900"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marcela Correia da Rocha","family":"Honorato","sequence":"additional","affiliation":[{"name":"Universidade Federal do Estado do Rio de Janeiro (UNIRIO) , , , ,","place":["Rio de Janeiro, RJ, Brazil, 20211-010"]},{"name":"Universidade do Estado do Rio de Janeiro (UERJ) , , , ,","place":["Rio de Janeiro, RJ, Brazil, 20551-030"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hana de Jesu\u0301s Galiza","family":"Gomes","sequence":"additional","affiliation":[{"name":"Universidade Federal do Estado do Rio de Janeiro (UNIRIO) , , , ,","place":["Rio de Janeiro, RJ, Brazil, 20211-010"]},{"name":"Universidade do Estado do Rio de Janeiro (UERJ) , , , ,","place":["Rio de Janeiro, RJ, Brazil, 20551-030"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pedro Paulo Saldanha","family":"Coimbra","sequence":"additional","affiliation":[{"name":"Universidade Federal do Estado do Rio de Janeiro (UNIRIO) , , , ,","place":["Rio de Janeiro, RJ, Brazil, 20211-010"]},{"name":"Universidade do Estado do Rio de Janeiro (UERJ) , , , ,","place":["Rio de Janeiro, RJ, Brazil, 20551-030"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nayara Ceci\u0301lia do Couto","family":"Guedes","sequence":"additional","affiliation":[{"name":"Fundac\u0327a\u0303o Oswaldo Cruz (FIOCRUZ) , , 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20551-030"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4254-4415","authenticated-orcid":true,"given":"Beatriz Rodrigues","family":"Canabarro","sequence":"additional","affiliation":[{"name":"Centro Brasileiro de Pesquisas Fi\u0301sicas (CBPF) , ,","place":["Rio de Janeiro, Brazil, 22290-180"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriele Fa\u0301tima","family":"de Souza","sequence":"additional","affiliation":[{"name":"Fundac\u0327a\u0303o Oswaldo Cruz (FIOCRUZ) , , ,","place":["Rio de Janeiro, Brazil, 21040-900"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thais Morais","family":"de Brito","sequence":"additional","affiliation":[{"name":"Fundac\u0327a\u0303o Oswaldo Cruz (FIOCRUZ) , , ,","place":["Rio de Janeiro, Brazil, 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Nanobiotechnol."},{"key":"2026100610483992600_cit4","doi-asserted-by":"publisher","DOI":"10.1016\/j.cis.2022.102640","article-title":"Nanoparticles: From\nSynthesis to Applications and Beyond","volume":"303","author":"Selmani","year":"2022","journal-title":"Adv. Colloid\nInterface Sci."},{"issue":"11","key":"2026100610483992600_cit5","doi-asserted-by":"publisher","DOI":"10.3390\/catal12111386","article-title":"Classification,\nSynthetic, and Characterization Approaches to Nanoparticles, and Their\nApplications in Various Fields of Nanotechnology: A Review","volume":"12","author":"Khan","year":"2022","journal-title":"Catalysts"},{"issue":"3","key":"2026100610483992600_cit6","doi-asserted-by":"publisher","first-page":"223","DOI":"10.1080\/17518253.2020.1802517","article-title":"Detail Review on Chemical,\nPhysical and Green Synthesis, Classification, Characterizations and\nApplications of Nanoparticles","volume":"13","author":"Ijaz","year":"2020","journal-title":"Green Chem. Lett.\nRev."},{"issue":"4","key":"2026100610483992600_cit7","doi-asserted-by":"publisher","DOI":"10.3390\/jfb13040274","article-title":"Metal Oxide Nanoparticles: Review of Synthesis, Characterization\nand Biological Effects","volume":"13","author":"Negrescu","year":"2022","journal-title":"J. Funct. Biomater."},{"issue":"17","key":"2026100610483992600_cit8","doi-asserted-by":"publisher","DOI":"10.3390\/nano12173066","article-title":"Current Research on Zinc Oxide Nanoparticles: Synthesis,\nCharacterization, and Biomedical Applications","volume":"12","author":"Mandal","year":"2022","journal-title":"Nanomaterials"},{"key":"2026100610483992600_cit9","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1155\/2018\/1062562","article-title":"The Advancing of Zinc Oxide Nanoparticles for Biomedical\nApplications","volume":"2018","author":"Jiang","year":"2018","journal-title":"Bioinorg. Chem. Appl."},{"issue":"4","key":"2026100610483992600_cit10","doi-asserted-by":"publisher","DOI":"10.3390\/pr11041193","article-title":"Zinc Oxide Nanoparticles: Synthesis,\nCharacterization, Modification, and Applications in Food and Agriculture","volume":"11","author":"Zhou","year":"2023","journal-title":"Processes"},{"issue":"10","key":"2026100610483992600_cit11","doi-asserted-by":"publisher","DOI":"10.3390\/nano15100754","article-title":"Zinc Oxide\nNanoparticles in Modern Science and Technology: Multifunctional Roles\nin Healthcare, Environmental Remediation, and Industry","volume":"15","author":"Lebaka","year":"2025","journal-title":"Nanomaterials"},{"issue":"9","key":"2026100610483992600_cit12","doi-asserted-by":"publisher","DOI":"10.3390\/nano13091500","article-title":"Recent Advances\nin ZnO Nanomaterial-Mediated Biological Applications and Action Mechanisms","volume":"13","author":"Xie","year":"2023","journal-title":"Nanomaterials"},{"issue":"17","key":"2026100610483992600_cit13","doi-asserted-by":"publisher","DOI":"10.3390\/w14172669","article-title":"Single and\nCombined Toxicity Effects of Zinc Oxide Nanoparticles: Uptake and\nAccumulation in Marine Microalgae, Toxicity Mechanisms, and Their\nFate in the Marine Environment","volume":"14","author":"Hazeem","year":"2022","journal-title":"Water"},{"issue":"4","key":"2026100610483992600_cit14","doi-asserted-by":"publisher","first-page":"300","DOI":"10.1080\/15376516.2018.1553221","article-title":"Zinc Oxide\nNanoparticles Impacts: Cytotoxicity, Genotoxicity, Developmental Toxicity,\nand Neurotoxicity","volume":"29","author":"Singh","year":"2019","journal-title":"Toxicol. Mech. Methods"},{"issue":"3","key":"2026100610483992600_cit15","doi-asserted-by":"publisher","DOI":"10.3390\/pharmaceutics17030281","article-title":"Nanotechnology to Overcome Blood\u2013Brain Barrier\nPermeability and Damage in Neurodegenerative Diseases","volume":"17","author":"Jime\u0301nez","year":"2025","journal-title":"Pharmaceutics"},{"issue":"1","key":"2026100610483992600_cit16","doi-asserted-by":"publisher","DOI":"10.1186\/s11671-016-1508-4","article-title":"Is Neurotoxicity of Metallic Nanoparticles the Cascades of Oxidative\nStress?","volume":"11","author":"Song","year":"2016","journal-title":"Nanoscale Res. Lett."},{"issue":"1","key":"2026100610483992600_cit17","doi-asserted-by":"publisher","DOI":"10.3390\/toxics14010011","article-title":"Zinc Oxide Nanoparticle-Induced Neurotoxicity:\nUnderlying Molecular Mechanisms and Future Perspectives","volume":"14","author":"Chen","year":"2026","journal-title":"Toxics"},{"issue":"9","key":"2026100610483992600_cit18","doi-asserted-by":"publisher","first-page":"5921","DOI":"10.1007\/s13204-023-02863-w","article-title":"Cosmetic Nanomaterials\nin the Environment: Nano-Zinc Oxide and Zinc-Influence on Soil Microorganisms","volume":"13","author":"Abdelmeged","year":"2023","journal-title":"Appl. Nanosci."},{"issue":"1","key":"2026100610483992600_cit19","doi-asserted-by":"publisher","first-page":"219","DOI":"10.1515\/mgmc-2022-0024","article-title":"Comparative Toxic Effect of Bulk\nZinc Oxide (ZnO) and ZnO Nanoparticles on Human Red Blood Cells","volume":"45","author":"Salami","year":"2022","journal-title":"Main Group Met. Chem."},{"key":"2026100610483992600_cit20","article-title":"Test No. 471:\nBacterial Reverse Mutation Test; OECD Guidelines for the Testing of\nChemicals, Section 4;","author":"OECD.","year":"2020"},{"key":"2026100610483992600_cit21","article-title":"Test No. 487:\nIn Vitro Mammalian Cell Micronucleus Test; OECD Guidelines for the\nTesting of Chemicals, Section 4;","author":"OECD.","year":"2023"},{"key":"2026100610483992600_cit22","doi-asserted-by":"publisher","DOI":"10.1787\/9789264203709-en","article-title":"Test No. 236:\nFish Embryo Acute Toxicity (FET) Test; OECD Guidelines for the Testing\nof Chemicals, Section 2;","author":"OECD.","year":"2025"},{"issue":"4","key":"2026100610483992600_cit23","doi-asserted-by":"publisher","first-page":"137","DOI":"10.1080\/10937404.2020.1751756","article-title":"A Review of Hepatic Nanotoxicology\n\u2013 Summation of Recent Findings and Considerations for the next\nGeneration of Study Designs","volume":"23","author":"Kermanizadeh","year":"2020","journal-title":"J. Toxicol. Environ.\nHeal. Part B"},{"key":"2026100610483992600_cit24","doi-asserted-by":"publisher","first-page":"391","DOI":"10.1016\/j.scitotenv.2017.06.125","article-title":"Uranium Mining\nWastes: The Use of the Fish Embryo Acute Toxicity Test (FET) Test\nto Evaluate Toxicity and Risk of Environmental Discharge","volume":"605\u2013606","author":"Lourenc\u0327o","year":"2017","journal-title":"Sci. Total Environ."},{"key":"2026100610483992600_cit25","doi-asserted-by":"publisher","DOI":"10.1016\/j.cbpc.2023.109701","article-title":"Toxic Effects of Polystyrene Nanoparticles\non the Development, Escape Locomotion, and Lateral-Line Sensory Function\nof Zebrafish Embryos","volume":"272","author":"Lin","year":"2023","journal-title":"Comp. Biochem. Physiol.\nPart C Toxicol. Pharmacol."},{"key":"2026100610483992600_cit26","doi-asserted-by":"publisher","DOI":"10.1016\/j.chemosphere.2023.140592","article-title":"Prediction of Acute Fish Toxicity\n(AFT) and Fish Embryo Toxicity (FET) Tests by Cytotoxicity Assays\nUsing Liver and Embryo Zebrafish Cell Lines (ZFL and ZEM2S)","volume":"346","author":"de Souza","year":"2024","journal-title":"Chemosphere"},{"key":"2026100610483992600_cit27","doi-asserted-by":"publisher","DOI":"10.1016\/j.aquatox.2023.106791","article-title":"Developmental Toxicity and Neurobehavioral Effects\nof Sodium Selenite and Selenium Nanoparticles on Zebrafish Embryos","volume":"266","author":"Hariharan","year":"2024","journal-title":"Aquat. Toxicol."},{"key":"2026100610483992600_cit28","doi-asserted-by":"publisher","DOI":"10.1016\/j.cbpc.2024.109838","article-title":"Assessing Cardiovascular Toxicity in Zebrafish Embryos\nExposed to Copper Nanoparticles","volume":"277","author":"Hsiao","year":"2024","journal-title":"Comp. Biochem.\nPhysiol. Part C Toxicol. Pharmacol."},{"issue":"7","key":"2026100610483992600_cit29","doi-asserted-by":"publisher","first-page":"676","DOI":"10.1038\/nmeth.2019","article-title":"Fiji: An Open-Source\nPlatform for Biological-Image Analysis","volume":"9","author":"Schindelin","year":"2012","journal-title":"Nat.\nMethods"},{"issue":"3\u20134","key":"2026100610483992600_cit30","doi-asserted-by":"publisher","first-page":"173","DOI":"10.1016\/0165-1161(83)90010-9","article-title":"Revised Methods for the Salmonella Mutagenicity Test","volume":"113","author":"Maron","year":"1983","journal-title":"Mutat. Res. Mutagen. Relat. 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