Folic Acid-Conjugated PEG Nanoparticles for Controlled Delivery of Fasudil in Experimental Autoimmune Encephalomyelitis: A Proposed Model for Effective Multiple Sclerosis Therapy

Folic Acid-Conjugated PEG Nanoparticles for Controlled Delivery of Fasudil in Experimental Autoimmune Encephalomyelitis: A Proposed Model for Effective Multiple Sclerosis Therapy | 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 Article Folic Acid-Conjugated PEG Nanoparticles for Controlled Delivery of Fasudil in Experimental Autoimmune Encephalomyelitis: A Proposed Model for Effective Multiple Sclerosis Therapy Niloufar Sehat, Seyed Massood Nabavi, Nasrin Lotfibakhshaiesh, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8475763/v1 This work is licensed under a CC BY 4.0 License Status: Under Revision Version 1 posted 10 You are reading this latest preprint version Abstract Background : Effective delivery of therapeutic agents is crucial in modern medicine, especially as conventional treatments face challenges such as limited bioavailability, adverse side effects, and frequent dosing requirements. This study focuses on the synthesis and characterization of PLGA-PEG-folic acid (FA) nanoparticles loaded with Fasudil, a Rho-associated kinase (ROCK) inhibitor, to improve Experimental Autoimmune Encephalomyelitis (EAE) in a mouse model of multiple sclerosis (MS). Methods : The nanoparticles were synthesized by conjugating folic acid to PLGA-PEG-NH₂, followed by nanoprecipitation for drug encapsulation. Comprehensive characterization assessed particle size, zeta potential, morphology, and encapsulation efficiency to ensure optimal performance and stability. Drug release studies were performed under varying pH conditions to evaluate release kinetics. Additionally, inflammatory cytokine levels, immunohistochemical markers including Glial Fibrillary Acidic Protein (GFAP), Ionized Calcium-Binding Adapter Molecule 1 (IBA1), Myelin Basic Protein (MBP), and 5-bromo-2′-deoxyuridine (BrdU) expression were measured alongside demyelination rates. Results: The PLGA-PEG-FA nanoparticles exhibited an average size of 252 ± 5 nm and a zeta potential of –24.2 mV, indicating good stability, with an encapsulation efficiency of 64.29% for Fasudil. Release profiles were pH-dependent and best described by the Korsmeyer-Peppas model. Clinically, mice treated with PLGA-PEG-FA nanoparticles showed reduced inflammatory cytokine levels, decreased expression of GFAP and IBA1, and increased expression of MBP and BrdU. Conclusions: These results demonstrate that PLGA-PEG-FA nanoparticles effectively encapsulate and release Fasudil in a controlled manner and significantly ameliorate EAE symptoms, highlighting their potential as an efficient drug delivery system for neuroinflammatory diseases such as MS. Biological sciences/Biochemistry Biological sciences/Drug discovery Health sciences/Neurology Biological sciences/Neuroscience Folic Acid PEG Nanoparticles Controlled Delivery Fasudil Multiple Sclerosis Experimental Autoimmune Encephalomyelitis (EAE) Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Revision Version 1 posted Editorial decision: Revision requested 24 Feb, 2026 Reviews received at journal 24 Feb, 2026 Reviewers agreed at journal 12 Feb, 2026 Reviews received at journal 05 Feb, 2026 Reviewers agreed at journal 04 Feb, 2026 Reviewers invited by journal 13 Jan, 2026 Editor assigned by journal 13 Jan, 2026 Editor invited by journal 12 Jan, 2026 Submission checks completed at journal 08 Jan, 2026 First submitted to journal 08 Jan, 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. 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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-8475763","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":573745351,"identity":"35284c9b-2839-4fd8-b5cb-bf5910b541ec","order_by":0,"name":"Niloufar Sehat","email":"","orcid":"","institution":"Royan Institute","correspondingAuthor":false,"prefix":"","firstName":"Niloufar","middleName":"","lastName":"Sehat","suffix":""},{"id":573745356,"identity":"d4bf4119-cd7c-4d11-ad04-d07e92895331","order_by":1,"name":"Seyed Massood 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Therapy","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"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":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Folic Acid, PEG Nanoparticles, Controlled Delivery, Fasudil, Multiple Sclerosis, Experimental Autoimmune Encephalomyelitis (EAE) ","lastPublishedDoi":"10.21203/rs.3.rs-8475763/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8475763/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground\u003c/strong\u003e: Effective delivery of therapeutic agents is crucial in modern medicine, especially as conventional treatments face challenges such as limited bioavailability, adverse side effects, and frequent dosing requirements. This study focuses on the synthesis and characterization of PLGA-PEG-folic acid (FA) nanoparticles loaded with Fasudil, a Rho-associated kinase (ROCK) inhibitor, to improve Experimental Autoimmune Encephalomyelitis (EAE) in a mouse model of multiple sclerosis (MS).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods\u003c/strong\u003e: The nanoparticles were synthesized by conjugating folic acid to PLGA-PEG-NH₂, followed by nanoprecipitation for drug encapsulation. Comprehensive characterization assessed particle size, zeta potential, morphology, and encapsulation efficiency to ensure optimal performance and stability. Drug release studies were performed under varying pH conditions to evaluate release kinetics. Additionally, inflammatory cytokine levels, immunohistochemical markers including Glial Fibrillary Acidic Protein (GFAP), Ionized Calcium-Binding Adapter Molecule 1 (IBA1), Myelin Basic Protein (MBP), and 5-bromo-2′-deoxyuridine (BrdU) expression were measured alongside demyelination rates.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults: \u003c/strong\u003eThe PLGA-PEG-FA nanoparticles exhibited an average size of 252 ± 5 nm and a zeta potential of –24.2 mV, indicating good stability, with an encapsulation efficiency of 64.29% for Fasudil. Release profiles were pH-dependent and best described by the Korsmeyer-Peppas model. Clinically, mice treated with PLGA-PEG-FA nanoparticles showed reduced inflammatory cytokine levels, decreased expression of GFAP and IBA1, and increased expression of MBP and BrdU.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions:\u003c/strong\u003e These results demonstrate that PLGA-PEG-FA nanoparticles effectively encapsulate and release Fasudil in a controlled manner and significantly ameliorate EAE symptoms, highlighting their potential as an efficient drug delivery system for neuroinflammatory diseases such as MS.\u003c/p\u003e","manuscriptTitle":"Folic Acid-Conjugated PEG Nanoparticles for Controlled Delivery of Fasudil in Experimental Autoimmune Encephalomyelitis: A Proposed Model for Effective Multiple Sclerosis Therapy","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-01-14 15:27:54","doi":"10.21203/rs.3.rs-8475763/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-02-24T08:27:58+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-02-24T07:57:32+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"268495621380122103469372695757917339308","date":"2026-02-12T11:30:30+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-02-06T00:02:14+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"25321803008551888819255458226360435684","date":"2026-02-04T15:59:40+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-01-13T08:01:20+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-01-13T07:49:51+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2026-01-12T13:05:51+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-01-08T09:43:03+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2026-01-08T09:22:05+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"b318bf72-2da4-4efb-9e2a-994c1888a21d","owner":[],"postedDate":"January 14th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"in-revision","subjectAreas":[{"id":61044651,"name":"Biological sciences/Biochemistry"},{"id":61044652,"name":"Biological sciences/Drug discovery"},{"id":61044653,"name":"Health sciences/Neurology"},{"id":61044654,"name":"Biological sciences/Neuroscience"}],"tags":[],"updatedAt":"2026-02-24T08:39:27+00:00","versionOfRecord":[],"versionCreatedAt":"2026-01-14 15:27:54","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8475763","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8475763","identity":"rs-8475763","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}