Towards Effective Radiation-Induced Hypothyroidism Therapeutics: Identifying Potential Biomarkers for In Vitro Thyroid Scaffold Evaluation

Towards Effective Radiation-Induced Hypothyroidism Therapeutics: Identifying Potential Biomarkers for In Vitro Thyroid Scaffold Evaluation | 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 Towards Effective Radiation-Induced Hypothyroidism Therapeutics: Identifying Potential Biomarkers for In Vitro Thyroid Scaffold Evaluation Maria Heim, Tara Fay, Daniel Grant, Elaine Emmerson, Anthony Callanan This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9303851/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 4 You are reading this latest preprint version Abstract Objective: Radiation-induced hypothyroidism (RIHT) affects up to 92% of head and neck cancer patients following irradiation,significantly impairing quality of life. Existing treatments show multiple limitations, but thyroid tissue engineering presents a promisingalternative. Validated biomarkers for assessing thyroid scaffolds in vitro remain limited. This study identifies candidate protein markersin the Nthy-ori 3-1 cell line, informed by expression patterns in a murine RIHT model. Methods: Immunofluorescent analysis of murine thyroids at days 3 and 28 post-irradiation identified preliminary protein candidates.An electrospun 10% (w/v) polycaprolactone scaffold was fabricated and characterized, then seeded with Nthy-ori 3-1 cells to monitorbiomarker expression over 14 days. Results: Imaging demonstrated reduced thyroxine in irradiated glands, confirming RIHT onset. Decreased E-Cadherin and increasedα-smooth-muscle actin and fibronectin indicated fibrosis, while transient Collagen-1 reduction reflected extracellular matrix remodelingwith reduced Zonula Occludens-1 (ZO-1) suggesting epithelial disruption. Within scaffolds, Nthy-ori 3-1 cells displayed a six-foldincrease in DNA content, higher metabolic activity, and collagen-like fibril formation. Expression changes in α-smooth-muscle actin,fibronectin, ZO-1, and Collagen-1 mirrorred irradiation-induced alterations but normalized by day 14, while increasing E-Cadherin andthyroid-specific proteins suggest emerging hormone-producing potential of the scaffold construct. Conclusion: These findings identified key biomarkers for evaluating thyroid scaffold performance in vitro and support their relevance forRIHT tissue-repair strategies. Full Text Additional Declarations No competing interests reported. Supplementary Files OnlineResource1.pptx OnlineResource2.pptx Cite Share Download PDF Status: Under Review Version 1 posted Reviewers invited by journal 07 May, 2026 Editor assigned by journal 06 Apr, 2026 Submission checks completed at journal 06 Apr, 2026 First submitted to journal 02 Apr, 2026 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. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-9303851","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":641425199,"identity":"fd1910c6-f73e-4c59-94e9-9d96d569e46a","order_by":0,"name":"Maria Heim","email":"","orcid":"","institution":"University of Edinburgh","correspondingAuthor":false,"prefix":"","firstName":"Maria","middleName":"","lastName":"Heim","suffix":""},{"id":641425200,"identity":"b6a3eecd-e446-4459-b2bb-45799d6ae463","order_by":1,"name":"Tara Fay","email":"","orcid":"","institution":"University of 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Devices","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-9303851/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9303851/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Objective: Radiation-induced hypothyroidism (RIHT) affects up to 92% of head and neck cancer patients following irradiation,significantly impairing quality of life. Existing treatments show multiple limitations, but thyroid tissue engineering presents a promisingalternative. Validated biomarkers for assessing thyroid scaffolds in vitro remain limited. This study identifies candidate protein markersin the Nthy-ori 3-1 cell line, informed by expression patterns in a murine RIHT model.\nMethods: Immunofluorescent analysis of murine thyroids at days 3 and 28 post-irradiation identified preliminary protein candidates.An electrospun 10% (w/v) polycaprolactone scaffold was fabricated and characterized, then seeded with Nthy-ori 3-1 cells to monitorbiomarker expression over 14 days.\nResults: Imaging demonstrated reduced thyroxine in irradiated glands, confirming RIHT onset. Decreased E-Cadherin and increasedα-smooth-muscle actin and fibronectin indicated fibrosis, while transient Collagen-1 reduction reflected extracellular matrix remodelingwith reduced Zonula Occludens-1 (ZO-1) suggesting epithelial disruption. Within scaffolds, Nthy-ori 3-1 cells displayed a six-foldincrease in DNA content, higher metabolic activity, and collagen-like fibril formation. 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