Efficacy of Netrin-1 Primed WJMSCs-Derived Secretome in Promoting Wound Healing: An In vitro Study

Efficacy of Netrin-1 Primed WJMSCs-Derived Secretome in Promoting Wound Healing: An In vitro Study | 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 Efficacy of Netrin-1 Primed WJMSCs-Derived Secretome in Promoting Wound Healing: An In vitro Study Shweta Verma, Jahnavy Madhukar Joshi, Raghavendra Upadhya, Samatha Bhat, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8132161/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 Wound healing is a multifaceted biological process that requires tightly regulated cellular activities, including proliferation, migration, and extracellular matrix remodeling. Mesenchymal stromal cells (MSCs) derived from Wharton’s Jelly (WJMSC) have emerged as powerful agents in regenerative medicine, largely attributed to their secretome, a dynamic mixture of bioactive molecules and extracellular vesicles. Recent advancements suggest that priming MSCs can potentiate their secretome’s therapeutic efficacy. However, the ability to selectively enhance wound repair mechanisms via tailored priming remains unexplored. In this study, we introduce recombinant netrin-1, a multifunctional guidance cue known for its dual role in angiogenesis and immune modulation, as a novel priming agent for WJMSCs. We demonstrate that netrin-1 priming significantly enriches the WJMSCs secretome, amplifying its regenerative potential. Conditioned media collected post 48 hours of netrin-1 PRIMING markedly enhanced fibroblast proliferation and migration, boosted angiogenic tube formation in HUVECs, and skewed macrophage polarization toward an anti-inflammatory M2 phenotype, required as key processes that orchestrates effective tissue repair. This study clearly establishes for the first time the role of netrin-1 priming in creating a functionally superior WJMSCs secretome that targets multiple facets of wound healing synergistically. Unlike conventional single-factor treatments, this approach leverages the natural versatility of MSC secretome augmented with a well-characterized bioactive cue, offering a sophisticated and cell-free therapeutic avenue. The netrin-1 WJMSC secretome presents a promising platform with translational potential to address complex wound healing challenges, especially chronic and non-healing wounds with persistent inflammation and impaired angiogenesis. Altogether, the N1-primed WJ-MSC secretome represents a promising, cell-free therapeutic strategy for enhancing wound healing and tissue regeneration. Future studies will focus on the development of targeted delivery systems to optimize their therapeutic efficacy in preclinical wound models, paving the way for advanced regenerative medicine interventions. Netrin-1 Mesenchymal Stromal Cell Secretome Priming Full Text Additional Declarations Competing interest reported. A patent has been filed by Manipal Academy of Higher Education for the optimization of the priming methodology (Application No. 202441071569). Supplementary Files GraphicalAbstract.png 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. 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-8132161","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":566867119,"identity":"024089f7-64a5-4853-b7b4-d65cd59918ae","order_by":0,"name":"Shweta Verma","email":"","orcid":"","institution":"Manipal Academy of Higher Education","correspondingAuthor":false,"prefix":"","firstName":"Shweta","middleName":"","lastName":"Verma","suffix":""},{"id":566867120,"identity":"a80ed416-ef5d-4e9f-8155-61d07c51838b","order_by":1,"name":"Jahnavy Madhukar Joshi","email":"","orcid":"","institution":"Manipal Academy of Higher 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A patent has been filed by Manipal Academy of Higher Education for the optimization of the priming methodology (Application No. 202441071569).","formattedTitle":"\u003cp\u003e\u003cstrong\u003eEfficacy of Netrin-1 Primed WJMSCs-Derived Secretome in Promoting Wound Healing: An \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eIn vitro\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e Study\u003c/strong\u003e\u003c/p\u003e","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"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":"Netrin-1, Mesenchymal Stromal Cell, Secretome, Priming","lastPublishedDoi":"10.21203/rs.3.rs-8132161/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8132161/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eWound healing is a multifaceted biological process that requires tightly regulated cellular activities, including proliferation, migration, and extracellular matrix remodeling. Mesenchymal stromal cells (MSCs) derived from Wharton’s Jelly (WJMSC) have emerged as powerful agents in regenerative medicine, largely attributed to their secretome, a dynamic mixture of bioactive molecules and extracellular vesicles. Recent advancements suggest that priming MSCs can potentiate their secretome’s therapeutic efficacy. However, the ability to selectively enhance wound repair mechanisms via tailored priming remains unexplored.\u003c/p\u003e\n\u003cp\u003eIn this study, we introduce recombinant netrin-1, a multifunctional guidance cue known for its dual role in angiogenesis and immune modulation, as a novel priming agent for WJMSCs. We demonstrate that netrin-1 priming significantly enriches the WJMSCs secretome, amplifying its regenerative potential. Conditioned media collected post 48 hours of netrin-1 PRIMING markedly enhanced fibroblast proliferation and migration, boosted angiogenic tube formation in HUVECs, and skewed macrophage polarization toward an anti-inflammatory M2 phenotype, required as key processes that orchestrates effective tissue repair.\u003c/p\u003e\n\u003cp\u003eThis study clearly establishes for the first time the role of netrin-1 priming in creating a functionally superior WJMSCs secretome that targets multiple facets of wound healing synergistically. Unlike conventional single-factor treatments, this approach leverages the natural versatility of MSC secretome augmented with a well-characterized bioactive cue, offering a sophisticated and cell-free therapeutic avenue. The netrin-1 WJMSC secretome presents a promising platform with translational potential to address complex wound healing challenges, especially chronic and non-healing wounds with persistent inflammation and impaired angiogenesis.\u003c/p\u003e\n\u003cp\u003eAltogether, the N1-primed WJ-MSC secretome represents a promising, cell-free therapeutic strategy for enhancing wound healing and tissue regeneration. Future studies will focus on the development of targeted delivery systems to optimize their therapeutic efficacy in preclinical wound models, paving the way for advanced regenerative medicine interventions.\u003c/p\u003e","manuscriptTitle":"Efficacy of Netrin-1 Primed WJMSCs-Derived Secretome in Promoting Wound Healing: An In vitro Study","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-12-31 08:40:19","doi":"10.21203/rs.3.rs-8132161/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","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}}],"origin":"","ownerIdentity":"1b4f659a-9ae4-4914-9541-250d27740f85","owner":[],"postedDate":"December 31st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-05-03T09:09:41+00:00","versionOfRecord":[],"versionCreatedAt":"2025-12-31 08:40:19","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8132161","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8132161","identity":"rs-8132161","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}