{"paper_id":"0767cc2c-e5ce-4209-9821-91c4b6a4786e","body_text":"Senomorphic Activity of a Novel Standardized Propolis Extract in Human Dermal Fibroblasts: Molecular Insights into Clinically Proven Anti-Wrinkle Efficacy | 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 Senomorphic Activity of a Novel Standardized Propolis Extract in Human Dermal Fibroblasts: Molecular Insights into Clinically Proven Anti-Wrinkle Efficacy Božo Radić, Jelena Šuran This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8572260/v2 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 18 Mar, 2026 Read the published version in Journal of Cosmetic Dermatology → Version 2 posted You are reading this latest preprint version Show more versions Abstract Background We recently demonstrated in a randomized controlled trial (RCT) that a Standardized Propolis Extract (SPE), produced via a patented non-alcoholic PEG 400/lecithin process, achieves significant clinical anti-wrinkle efficacy (34% wrinkle depth reduction). The present study investigates the underlying molecular mechanisms, specifically its potential senomorphic activity—the ability to modulate the Senescence-Associated Secretory Phenotype (SASP) without inducing cell death. Objective To evaluate the senomorphic activity of this chemically defined SPE (standardized to 1318.43 µg/g total phenolic markers) in an in vitro model of oxidative stress-induced senescence, providing molecular insights into its clinically observed anti-aging effects. Methods Human Dermal Fibroblasts (HDFs) were pre-treated with SPE (0.01%, 0.05%) or Rapamycin (3µM, reference senomorphic control). Senescence was induced via a validated stress-induced premature senescence (SIPS) protocol (200µM H₂O₂, 2 hours). Gene expression for senescence markers (CDKN2A/p16, CDKN1A/p21), SASP cytokines (IL-6, IL-8), and cell cycle regulators (CDK4, CDK2, CCNE1) was quantified by qPCR. An exploratory study on Mesenchymal Stem Cells (MSCs) assessed SA-β-galactosidase activity qualitatively. Results The 0.05% SPE demonstrated potent senomorphic activity, significantly suppressing the key SASP marker IL-6 (FC: -7.78, p = 0.003)—comparable to the Rapamycin control (FC: -8.1, p = 0.003). Uniquely, SPE induced transcriptional upregulation of CDK4 (FC: +6.71, p = 0.002) and CDKN1A/p21 (FC: +2.33, p = 0.005), effects not observed with Rapamycin. In exploratory MSC experiments, SPE qualitatively reduced SA-β-gal staining. Conclusion This first-in-class standardized propolis extract demonstrates distinct senomorphic activity, suppressing the inflammatory SASP (IL-6) while inducing transcriptional modulation of pro-regenerative pathways (CDK4). These molecular findings provide mechanistic insights consistent with the extract's clinically proven anti-wrinkle efficacy, supporting its positioning as an evidence-based active ingredient for dermo-cosmetic formulations targeting inflammaging. Dermatology Cell Survival and Cell Death Cell Cycle & Proliferation propolis standardized extract senomorphic cellular senescence inflammaging SASP IL-6 CDK4 human dermal fibroblasts Full Text Additional Declarations The authors declare potential competing interests as follows: Božo Radić and Jelena Šuran are employees and shareholders of Apiotix Technologies d.o.o., the company that holds the internationally granted patent (WO/2020/169425, with patents granted in the European Union [EP2020706145], United States [US17430567], China [CN202080015084.X], India [IN202117037021], Eurasia [EA202192022], and Mexico [MX/a/2021/009877]) for the standardized propolis extract evaluated in this study. Cite Share Download PDF Status: Published Journal Publication published 18 Mar, 2026 Read the published version in Journal of Cosmetic Dermatology → Version 2 posted You are reading this latest preprint version Show more versions 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. 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study.\",\"formattedTitle\":\"\\u003cp\\u003e\\u003cstrong\\u003eSenomorphic Activity of a Novel Standardized Propolis Extract in Human Dermal Fibroblasts: Molecular Insights into Clinically Proven Anti-Wrinkle Efficacy\\u003c/strong\\u003e\\u003c/p\\u003e\",\"fulltext\":[],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":false,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":true,\"hideJournal\":false,\"highlight\":\"\",\"institution\":\"\",\"isAcceptedByJournal\":true,\"isAuthorSuppliedPdf\":true,\"isDeskRejected\":\"\",\"isHiddenFromSearch\":false,\"isInQc\":false,\"isInWorkflow\":false,\"isPdf\":true,\"isPdfUpToDate\":true,\"isWithdrawnOrRetracted\":false,\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"researchsquare\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":true,\"externalIdentity\":\"\",\"sideBox\":\"\",\"snPcode\":\"\",\"submissionUrl\":\"/submission\",\"title\":\"Research 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The present study investigates the underlying molecular mechanisms, specifically its potential senomorphic activity\\u0026mdash;the ability to modulate the Senescence-Associated Secretory Phenotype (SASP) without inducing cell death.\\u003c/p\\u003e\\u003ch2\\u003eObjective\\u003c/h2\\u003e \\u003cp\\u003eTo evaluate the senomorphic activity of this chemically defined SPE (standardized to 1318.43 \\u0026micro;g/g total phenolic markers) in an in vitro model of oxidative stress-induced senescence, providing molecular insights into its clinically observed anti-aging effects.\\u003c/p\\u003e\\u003ch2\\u003eMethods\\u003c/h2\\u003e \\u003cp\\u003eHuman Dermal Fibroblasts (HDFs) were pre-treated with SPE (0.01%, 0.05%) or Rapamycin (3\\u0026micro;M, reference senomorphic control). Senescence was induced via a validated stress-induced premature senescence (SIPS) protocol (200\\u0026micro;M H₂O₂, 2 hours). Gene expression for senescence markers (CDKN2A/p16, CDKN1A/p21), SASP cytokines (IL-6, IL-8), and cell cycle regulators (CDK4, CDK2, CCNE1) was quantified by qPCR. An exploratory study on Mesenchymal Stem Cells (MSCs) assessed SA-β-galactosidase activity qualitatively.\\u003c/p\\u003e\\u003ch2\\u003eResults\\u003c/h2\\u003e \\u003cp\\u003eThe 0.05% SPE demonstrated potent senomorphic activity, significantly suppressing the key SASP marker IL-6 (FC: -7.78, p\\u0026thinsp;=\\u0026thinsp;0.003)\\u0026mdash;comparable to the Rapamycin control (FC: -8.1, p\\u0026thinsp;=\\u0026thinsp;0.003). Uniquely, SPE induced transcriptional upregulation of CDK4 (FC: +6.71, p\\u0026thinsp;=\\u0026thinsp;0.002) and CDKN1A/p21 (FC: +2.33, p\\u0026thinsp;=\\u0026thinsp;0.005), effects not observed with Rapamycin. In exploratory MSC experiments, SPE qualitatively reduced SA-β-gal staining.\\u003c/p\\u003e\\u003ch2\\u003eConclusion\\u003c/h2\\u003e \\u003cp\\u003eThis first-in-class standardized propolis extract demonstrates distinct senomorphic activity, suppressing the inflammatory SASP (IL-6) while inducing transcriptional modulation of pro-regenerative pathways (CDK4). These molecular findings provide mechanistic insights consistent with the extract's clinically proven anti-wrinkle efficacy, supporting its positioning as an evidence-based active ingredient for dermo-cosmetic formulations targeting inflammaging.\\u003c/p\\u003e\",\"manuscriptTitle\":\"Senomorphic Activity of a Novel Standardized Propolis Extract in Human Dermal Fibroblasts: Molecular Insights into Clinically Proven Anti-Wrinkle Efficacy\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":2,\"date\":\"2026-01-26 15:09:07\",\"doi\":\"10.21203/rs.3.rs-8572260/v2\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"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}},{\"code\":1,\"date\":\"2026-01-13 10:02:52\",\"doi\":\"10.21203/rs.3.rs-8572260/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"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\":\"8b9ec66e-a830-40ed-bd19-ed6a59ab3294\",\"owner\":[],\"postedDate\":\"January 26th, 2026\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"published-in-journal\",\"subjectAreas\":[{\"id\":61054191,\"name\":\"Dermatology\"},{\"id\":61054192,\"name\":\"Cell Survival and Cell Death\"},{\"id\":61054193,\"name\":\"Cell Cycle \\u0026 Proliferation\"}],\"tags\":[],\"updatedAt\":\"2026-03-23T22:16:43+00:00\",\"versionOfRecord\":{\"articleIdentity\":\"rs-8572260\",\"link\":\"https://doi.org/10.1111/jocd.70689\",\"journal\":{\"identity\":\"journal-of-cosmetic-dermatology\",\"isVorOnly\":true,\"title\":\"Journal of Cosmetic Dermatology\"},\"publishedOn\":\"2026-03-19 00:00:00\",\"publishedOnDateReadable\":\"March 19th, 2026\"},\"versionCreatedAt\":\"2026-01-26 15:09:07\",\"video\":\"\",\"vorDoi\":\"10.1111/jocd.70689\",\"vorDoiUrl\":\"https://doi.org/10.1111/jocd.70689\",\"workflowStages\":[]},\"version\":\"v2\",\"identity\":\"rs-8572260\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-8572260\",\"identity\":\"rs-8572260\",\"version\":[\"v2\"]},\"buildId\":\"XKTyCvWXoU3ODBz1xrDgd\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}