Wound Healing Effect of Dendrimer-Based Nano-Fluconazole/Nano-Chitosan on Cutaneous Leishmaniasis in Mice: Insights into the Mechanism of Action via NMR-Based Metabolomics

Wound Healing Effect of Dendrimer-Based Nano-Fluconazole/Nano-Chitosan on Cutaneous Leishmaniasis in Mice: Insights into the Mechanism of Action via NMR-Based Metabolomics | 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 Wound Healing Effect of Dendrimer-Based Nano-Fluconazole/Nano-Chitosan on Cutaneous Leishmaniasis in Mice: Insights into the Mechanism of Action via NMR-Based Metabolomics Homeyra Fadaei, Taher Elmi, Mohammad Taghi Goodarzi, Ali Kalantari Hesari This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7865741/v1 This work is licensed under a CC BY 4.0 License Status: Under Revision Version 1 posted 12 You are reading this latest preprint version Abstract Background Cutaneous leishmaniasis (CL), mainly caused by Leishmania major ( L. Major ), poses significant therapeutic challenges due to drug resistance, and adverse effects of conventional treatments. This study developed a dendrimer-based nanostructure combining fluconazole, an antileishmanial and antifungal agent, with chitosan, a biocompatible polymer exhibiting antileishmanial, antibacterial, and wound-healing properties. NMR-based metabolomics was employed to elucidate the underlying mechanisms of action. Methods A dendrimer was synthesized and conjugated with chitosan and fluconazole. Nanostructures were characterized by DLS, SEM, and FTIR, and evaluated for drug loading, solubility, and release kinetics. Anti-leishmanial efficacy was assessed in vitro against L. major promastigotes and in vivo in BALB/c mice through lesion size monitoring. Metabolic profiling via NMR analyzed drug/parasite interactions, while cytotoxicity and systemic toxicity were evaluated using macrophage assays, hematological, biochemical, and histopathological analyses. Results The nanostructure exhibited nanoscale size, uniform morphology, and efficient drug conjugation, with loading efficiencies of 49% (chitosan), and 56% (fluconazole). Encapsulation enhanced solubility and enabled sustained release. The NFLZ/Chi formulation demonstrated potent in vitro activity (IC₅₀ = 13 µg/mL) with synergistic effects (CI = 0.61) and achieved 97% wound healing in vivo. Metabolomics revealed significant disruption of amino acid, carbohydrate, and lipid metabolism, particularly branched-chain amino acids and glycine/serine/threonine pathways. Toxicity evaluations confirmed high biocompatibility and minimal hepatic alterations. Conclusion This dendrimer-based nanostructure effectively targets L. major , combining direct anti-parasitic effects with metabolic disruption, improved drug delivery, and accelerated wound healing, while exhibiting minimal toxicity. These results support its potential as a safe and efficient therapeutic strategy for CL. Biological sciences/Biochemistry Biological sciences/Biotechnology Physical sciences/Chemistry Biological sciences/Drug discovery Physical sciences/Nanoscience and technology Nano dendrimer Fluconazole Chitosan NMR Metabolomics Anti-Leishmanial Therapy Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Revision Version 1 posted Editorial decision: Revision requested 12 Dec, 2025 Reviews received at journal 01 Dec, 2025 Reviews received at journal 29 Nov, 2025 Reviews received at journal 24 Nov, 2025 Reviewers agreed at journal 08 Nov, 2025 Reviewers agreed at journal 06 Nov, 2025 Reviewers agreed at journal 04 Nov, 2025 Reviewers invited by journal 04 Nov, 2025 Editor assigned by journal 02 Nov, 2025 Editor invited by journal 29 Oct, 2025 Submission checks completed at journal 28 Oct, 2025 First submitted to journal 26 Oct, 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. 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-7865741","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":542329064,"identity":"c974f0f2-c5a1-42c0-9cb2-8fc03900b3d7","order_by":0,"name":"Homeyra Fadaei","email":"","orcid":"","institution":"Sha.C, Islamic Azad University","correspondingAuthor":false,"prefix":"","firstName":"Homeyra","middleName":"","lastName":"Fadaei","suffix":""},{"id":542329066,"identity":"c2a067e2-46f7-4db2-989a-a62ba9b442af","order_by":1,"name":"Taher 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Metabolomics","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":"Nano dendrimer, Fluconazole, Chitosan, NMR Metabolomics, Anti-Leishmanial Therapy","lastPublishedDoi":"10.21203/rs.3.rs-7865741/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7865741/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e\u003cp\u003eCutaneous leishmaniasis (CL), mainly caused by \u003cem\u003eLeishmania major\u003c/em\u003e (\u003cem\u003eL. Major\u003c/em\u003e), poses significant therapeutic challenges due to drug resistance, and adverse effects of conventional treatments. This study developed a dendrimer-based nanostructure combining fluconazole, an antileishmanial and antifungal agent, with chitosan, a biocompatible polymer exhibiting antileishmanial, antibacterial, and wound-healing properties. NMR-based metabolomics was employed to elucidate the underlying mechanisms of action.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e\u003cp\u003eA dendrimer was synthesized and conjugated with chitosan and fluconazole. Nanostructures were characterized by DLS, SEM, and FTIR, and evaluated for drug loading, solubility, and release kinetics. Anti-leishmanial efficacy was assessed in vitro against \u003cem\u003eL. major\u003c/em\u003e promastigotes and in vivo in BALB/c mice through lesion size monitoring. Metabolic profiling via NMR analyzed drug/parasite interactions, while cytotoxicity and systemic toxicity were evaluated using macrophage assays, hematological, biochemical, and histopathological analyses.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e\u003cp\u003eThe nanostructure exhibited nanoscale size, uniform morphology, and efficient drug conjugation, with loading efficiencies of 49% (chitosan), and 56% (fluconazole). Encapsulation enhanced solubility and enabled sustained release. The NFLZ/Chi formulation demonstrated potent in vitro activity (IC₅₀ = 13 \u0026micro;g/mL) with synergistic effects (CI\u0026thinsp;=\u0026thinsp;0.61) and achieved 97% wound healing in vivo. Metabolomics revealed significant disruption of amino acid, carbohydrate, and lipid metabolism, particularly branched-chain amino acids and glycine/serine/threonine pathways. Toxicity evaluations confirmed high biocompatibility and minimal hepatic alterations.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e\u003cp\u003eThis dendrimer-based nanostructure effectively targets \u003cem\u003eL. major\u003c/em\u003e, combining direct anti-parasitic effects with metabolic disruption, improved drug delivery, and accelerated wound healing, while exhibiting minimal toxicity. These results support its potential as a safe and efficient therapeutic strategy for CL.\u003c/p\u003e","manuscriptTitle":"Wound Healing Effect of Dendrimer-Based Nano-Fluconazole/Nano-Chitosan on Cutaneous Leishmaniasis in Mice: Insights into the Mechanism of Action via NMR-Based Metabolomics","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-13 17:22:16","doi":"10.21203/rs.3.rs-7865741/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-12-12T18:40:14+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-12-01T07:04:32+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-11-29T06:35:46+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-11-24T12:33:30+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"326954795712610217107614840698448756189","date":"2025-11-08T14:18:26+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"274990762457864514229512597445720839221","date":"2025-11-06T09:43:35+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"226673813165966168720868695155390301547","date":"2025-11-04T17:19:09+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-11-04T09:18:22+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-11-02T18:48:39+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-10-29T16:20:14+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-10-29T00:39:24+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2025-10-26T10:02:15+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":"442b1736-9c91-4ab2-9234-da612fa751c9","owner":[],"postedDate":"November 13th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"in-revision","subjectAreas":[{"id":57689497,"name":"Biological sciences/Biochemistry"},{"id":57689498,"name":"Biological sciences/Biotechnology"},{"id":57689500,"name":"Physical sciences/Chemistry"},{"id":57689502,"name":"Biological sciences/Drug discovery"},{"id":57689503,"name":"Physical sciences/Nanoscience and technology"}],"tags":[],"updatedAt":"2026-05-12T17:54:55+00:00","versionOfRecord":[],"versionCreatedAt":"2025-11-13 17:22:16","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7865741","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7865741","identity":"rs-7865741","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}