Enhanced biofilm inhibition via EPS-binding liposomes

Enhanced biofilm inhibition via EPS-binding liposomes | 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 Enhanced biofilm inhibition via EPS-binding liposomes Sayma Afrin, Tilahun Ayane Debele, Yoonjee Catherine Park This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7106450/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 05 Nov, 2025 Read the published version in Korean Journal of Chemical Engineering → Version 1 posted 4 You are reading this latest preprint version Abstract Treating infectious diseases with current available antimicrobial drugs is extremely difficult due to biofilms that act as barriers and reduce the concentration of antimicrobial agents that reach the bacteria embedded in the biofilms. In this study, we hypothesized that extracellular polymeric substances (EPS)-binding PEGylated liposomes anchor to biofilm matrices and sterically block the communication between bacteria, leading to biofilm inhibition. A 16-mer peptide, which binds to hyaluronic acid as one of the EPS, was covalently conjugated to PEG (polyethylene glycol)-lipid for producing EPS-binding liposomes. The effect of the liposomes on inhibiting or eradicating biofilm formation was investigated, compared to the PEGylated liposomes (bare PEGylated liposomes). Dynamic light scattering (DLS) measurement results showed that the EPS-binding PEGylated liposomes and bare PEGylated liposomes have a particle size of < 200nm and nearly neutral zeta potential. The molecular interaction of EPS extracted from S. aureus biofilm with EPS-binding liposomes and free EPS-binding peptides was determined using isothermal titration calorimetry (ITC) and the result revealed that EPS-binding liposome (Ka ~ 4.82×10 5 ) has better affinity than the free EPS-binding peptides (Ka ~ 1.79×10 3 ). The minimal biofilm inhibitory concentration (MBIC) assay showed EPS-binding liposomes have a better biofilm inhibition effect, in a dose-dependent manner, compared to the bare PEGylated liposomes and free EPS-binding peptides. Physical disruption and blocking chemical communication (quorum sensing (QS)) via biofilm binding are likely a key mechanism behind the effectiveness of EPS-binding liposomes in biofilm inhibition although further study is needed. Biofilm EPS-binding peptides hyaluronic acid peptide modified PEGylated liposome methicillin-resistant S. aureus (MRSA) Full Text Cite Share Download PDF Status: Published Journal Publication published 05 Nov, 2025 Read the published version in Korean Journal of Chemical Engineering → Version 1 posted Reviewers agreed at journal 18 Jul, 2025 Reviewers invited by journal 16 Jul, 2025 Editor assigned by journal 15 Jul, 2025 First submitted to journal 11 Jul, 2025 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-7106450","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":486597181,"identity":"1da1b73e-640f-45c3-8e25-5435dee34103","order_by":0,"name":"Sayma Afrin","email":"","orcid":"","institution":"University of Cincinnati","correspondingAuthor":false,"prefix":"","firstName":"Sayma","middleName":"","lastName":"Afrin","suffix":""},{"id":486597182,"identity":"ee91ec8e-b952-44b7-af1c-bb1ce4a9dac7","order_by":1,"name":"Tilahun Ayane Debele","email":"","orcid":"","institution":"University of Cincinnati","correspondingAuthor":false,"prefix":"","firstName":"Tilahun","middleName":"Ayane","lastName":"Debele","suffix":""},{"id":486597183,"identity":"0e74f39f-93c9-4a11-8544-e7412f900eeb","order_by":2,"name":"Yoonjee Catherine Park","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA20lEQVRIiWNgGAWjYHACZiC2AbMOPICIGBCjJQ2iJYEELYchTKK0mLOffWzMU3E+sX/a4YdAWw7LM7A3b5PAp8WyJ904mefM7cQZt9MMQFoMG3iOleHVYnAgjfkwb9vtxA3SCWAtCQwSOWb4tZx/BtTy7xxQS/oHiBb5NwS03EhjTuZtOADUkgOzhQe/FssZz5gN5xxLNp5xO6fgQIJBumEbT1qxBT4t5vxpzBJvauxk+2enb/7wocJanp/98MYbeB0GxEw8yFw2fMphWhh/EFI1CkbBKBgFIxsAAN8QSMJ8n4N9AAAAAElFTkSuQmCC","orcid":"","institution":"University of Cincinnati","correspondingAuthor":true,"prefix":"","firstName":"Yoonjee","middleName":"Catherine","lastName":"Park","suffix":""}],"badges":[],"createdAt":"2025-07-12 07:31:55","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7106450/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7106450/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s11814-025-00583-1","type":"published","date":"2025-11-05T15:57:38+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":95564053,"identity":"e33af0b9-5049-4f12-98a2-752d258ed2a0","added_by":"auto","created_at":"2025-11-10 16:07:02","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":746886,"visible":true,"origin":"","legend":"","description":"","filename":"07112025BiofilmManuscriptYP.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7106450/v1_covered_d57818e5-892a-4dae-80c9-2e9a14fb1e73.pdf"}],"financialInterests":"","formattedTitle":"Enhanced biofilm inhibition via EPS-binding liposomes","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"korean-journal-of-chemical-engineering","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"kjce","sideBox":"Learn more about [Korean Journal of Chemical Engineering](http://link.springer.com/journal/11814)","snPcode":"11814","submissionUrl":"https://www.editorialmanager.com/kjce/default2.aspx","title":"Korean Journal of Chemical Engineering","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Subscription","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Biofilm, EPS-binding peptides, hyaluronic acid, peptide modified PEGylated liposome, methicillin-resistant S. aureus (MRSA)","lastPublishedDoi":"10.21203/rs.3.rs-7106450/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7106450/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eTreating infectious diseases with current available antimicrobial drugs is extremely difficult due to biofilms that act as barriers and reduce the concentration of antimicrobial agents that reach the bacteria embedded in the biofilms. In this study, we hypothesized that extracellular polymeric substances (EPS)-binding PEGylated liposomes anchor to biofilm matrices and sterically block the communication between bacteria, leading to biofilm inhibition. A 16-mer peptide, which binds to hyaluronic acid as one of the EPS, was covalently conjugated to PEG (polyethylene glycol)-lipid for producing EPS-binding liposomes. The effect of the liposomes on inhibiting or eradicating biofilm formation was investigated, compared to the PEGylated liposomes (bare PEGylated liposomes). Dynamic light scattering (DLS) measurement results showed that the EPS-binding PEGylated liposomes and bare PEGylated liposomes have a particle size of \u0026lt;\u0026thinsp;200nm and nearly neutral zeta potential. The molecular interaction of EPS extracted from S. aureus biofilm with EPS-binding liposomes and free EPS-binding peptides was determined using isothermal titration calorimetry (ITC) and the result revealed that EPS-binding liposome (Ka\u0026thinsp;~\u0026thinsp;4.82\u0026times;10\u003csup\u003e5\u003c/sup\u003e) has better affinity than the free EPS-binding peptides (Ka\u0026thinsp;~\u0026thinsp;1.79\u0026times;10\u003csup\u003e3\u003c/sup\u003e). The minimal biofilm inhibitory concentration (MBIC) assay showed EPS-binding liposomes have a better biofilm inhibition effect, in a dose-dependent manner, compared to the bare PEGylated liposomes and free EPS-binding peptides. Physical disruption and blocking chemical communication (quorum sensing (QS)) via biofilm binding are likely a key mechanism behind the effectiveness of EPS-binding liposomes in biofilm inhibition although further study is needed.\u003c/p\u003e","manuscriptTitle":"Enhanced biofilm inhibition via EPS-binding liposomes","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-07-21 13:55:28","doi":"10.21203/rs.3.rs-7106450/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewerAgreed","content":"","date":"2025-07-18T15:51:33+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-07-17T02:07:17+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-07-15T15:25:49+00:00","index":"","fulltext":""},{"type":"submitted","content":"Korean Journal of Chemical Engineering","date":"2025-07-12T03:31:42+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"korean-journal-of-chemical-engineering","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"kjce","sideBox":"Learn more about [Korean Journal of Chemical Engineering](http://link.springer.com/journal/11814)","snPcode":"11814","submissionUrl":"https://www.editorialmanager.com/kjce/default2.aspx","title":"Korean Journal of Chemical Engineering","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Subscription","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"579627c4-e32a-4e5e-a5f4-74a5e5b5a820","owner":[],"postedDate":"July 21st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-11-10T16:01:33+00:00","versionOfRecord":{"articleIdentity":"rs-7106450","link":"https://doi.org/10.1007/s11814-025-00583-1","journal":{"identity":"korean-journal-of-chemical-engineering","isVorOnly":false,"title":"Korean Journal of Chemical Engineering"},"publishedOn":"2025-11-05 15:57:38","publishedOnDateReadable":"November 5th, 2025"},"versionCreatedAt":"2025-07-21 13:55:28","video":"","vorDoi":"10.1007/s11814-025-00583-1","vorDoiUrl":"https://doi.org/10.1007/s11814-025-00583-1","workflowStages":[]},"version":"v1","identity":"rs-7106450","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7106450","identity":"rs-7106450","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}