Metabolomic Responses of Osteoblasts to Zirconia Nanoparticles Across Dose and Time

Metabolomic Responses of Osteoblasts to Zirconia Nanoparticles Across Dose and Time | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Metabolomic Responses of Osteoblasts to Zirconia Nanoparticles Across Dose and Time Yuki Nakamura, Sakura Mori, Hiroshi Tanaka This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8963037/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract In this study, we investigated the dose- and time-dependent metabolic responses of human osteoblast-like cells following exposure to zirconia nanoparticles ZrO₂ NPs. Cells were treated with 0 10 25 50 and 100 µg mL ZrO₂ NPs for 24 and 72 h and analyzed using targeted liquid chromatography mass spectrometry based metabolomics. Cell viability reactive oxygen species production alkaline phosphatase activity and mineralized nodule formation were systematically evaluated. Cell viability remained above 90 percent at concentrations up to 50 µg mL whereas metabolomic analysis revealed significant alterations in glycolytic flux tricarboxylic acid cycle intermediates and glutathione metabolism with a false discovery rate below 0.05. Exposure to 100 µg mL ZrO₂ NPs led to increased oxidative stress reduced alkaline phosphatase activity and impaired mineralization. Partial least squares discriminant analysis demonstrated clear metabolic separation between control and treated groups with cysteine glutamate and citrate identified as key discriminatory metabolites. These results indicate that ZrO₂ NPs induce measurable metabolic modulation at sub toxic concentrations and affect osteogenic and redox related pathways in a concentration and time dependent manner. Dentistry zirconia nanoparticles osteoblasts LC–MS metabolomics oxidative stress mineralization Full Text Additional Declarations The authors declare no competing interests. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8963037","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":596652647,"identity":"e96534c2-dd02-4605-91a0-c9f57e8eaed8","order_by":0,"name":"Yuki Nakamura","email":"data:image/png;base64,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","orcid":"","institution":"","correspondingAuthor":true,"prefix":"","firstName":"Yuki","middleName":"","lastName":"Nakamura","suffix":""},{"id":596652648,"identity":"32bb0c88-dc56-43e2-a8f3-f81d46adf2be","order_by":1,"name":"Sakura Mori","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Sakura","middleName":"","lastName":"Mori","suffix":""},{"id":596652649,"identity":"33c8e2c4-85db-4aef-a5aa-3a4e8b18a873","order_by":2,"name":"Hiroshi Tanaka","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Hiroshi","middleName":"","lastName":"Tanaka","suffix":""}],"badges":[],"createdAt":"2026-02-25 04:32:51","currentVersionCode":1,"declarations":{"humanSubjects":false,"vertebrateSubjects":false,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false},"doi":"10.21203/rs.3.rs-8963037/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8963037/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":103533532,"identity":"07c82876-b59c-4fcf-bcea-655c8af46d47","added_by":"auto","created_at":"2026-02-26 17:46:53","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":520518,"visible":true,"origin":"","legend":"","description":"","filename":"11.27YeMingfu05MetabolomicSignaturesofOsteoblastResponsetoZirconiaNanoparticlesDoseandTimedependentEffects.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8963037/v1_covered_4be3304d-9860-4edf-ae4a-f13a0e13758b.pdf"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003e\u003cstrong\u003eMetabolomic Responses of Osteoblasts to Zirconia Nanoparticles Across Dose and Time\u003c/strong\u003e\u003c/p\u003e","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"zirconia nanoparticles, osteoblasts, LC–MS metabolomics, oxidative stress, mineralization","lastPublishedDoi":"10.21203/rs.3.rs-8963037/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8963037/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eIn this study, we investigated the dose- and time-dependent metabolic responses of human osteoblast-like cells following exposure to zirconia nanoparticles ZrO₂ NPs. 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