Fabrication and characterization of polymeric nanoparticle for efficient and sustained delivery of Acyclovir for antiviral therapy and in-silico study using Pk-Sim software

Fabrication and characterization of polymeric nanoparticle for efficient and sustained delivery of Acyclovir for antiviral therapy and in-silico study using Pk-Sim software | 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 Fabrication and characterization of polymeric nanoparticle for efficient and sustained delivery of Acyclovir for antiviral therapy and in-silico study using Pk-Sim software Sudipta Das, Arnab Samanta, Sourish Sarkar, Baishali Ghosh, Sawan Das, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7885636/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 10 Mar, 2026 Read the published version in Journal of Pharmaceutical Innovation → Version 1 posted You are reading this latest preprint version Abstract Purpose The aim of this study is to develop the nanoparticles of the weakly water-soluble antiviral medication Acyclovir using biodegradable and biocompatible natural polymer to overcome it’ s poor oral bioavailability and frequent dosing frequency were the goals of this work. Methods The ionic gelation method was used to prepare acyclovir loaded chitosan nanoparticle using STPP (Sodium tripolyphosphate) as the crosslinking agent. The formulations were characterized by determining particle size, surface morphology, percentage of yield, entrapment efficiency, drug- excipient compatibility (FT-IR), in-vitro release study and in-silico study. Results The FT-IR shows no incompatibility between the drug and the excipients. The analysis of dynamic light scattering indicated that nanoparticles were with it’s nanometric range (within 100 nm), demonstrating moderate polydispersity and SEM shows spherical morphology with smooth surface. The entrapment efficiency showed ⁓98% which indicates that chitosan is a potential carrier for the acyclovir. In-vitro release study illustrates sustain drug release over the time period and drug release kinetic best fitted in korsemeyer-peppas model which indicates diffusion-controlled release of the drug. Additionally, in-silico PBPK model predicted improved plasma profile compared to the conventional oral dosage form. Conclusion The findings suggested that the optimized formulation, especially F3, holding promise for sustained and controlled delivery of Acyclovir, potentially reducing dosing frequency and minimizing side effects. Nanoparticle STPP Ionic-Gelation FT-IR SEM PBPK Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 10 Mar, 2026 Read the published version in Journal of Pharmaceutical Innovation → 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-7885636","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":543066392,"identity":"37300946-65dd-4114-bd06-87715419b008","order_by":0,"name":"Sudipta Das","email":"data:image/png;base64,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","orcid":"","institution":"Netaji Subhas Chandra Bose Institute of Pharmacy","correspondingAuthor":true,"prefix":"","firstName":"Sudipta","middleName":"","lastName":"Das","suffix":""},{"id":543066393,"identity":"68374d4d-3aa9-412f-aad0-250d3f19bde9","order_by":1,"name":"Arnab Samanta","email":"","orcid":"","institution":"Netaji Subhas Chandra Bose Institute of Pharmacy","correspondingAuthor":false,"prefix":"","firstName":"Arnab","middleName":"","lastName":"Samanta","suffix":""},{"id":543066394,"identity":"611126d3-4702-406f-b497-58deddb96818","order_by":2,"name":"Sourish Sarkar","email":"","orcid":"","institution":"Netaji Subhas Chandra Bose Institute of Pharmacy","correspondingAuthor":false,"prefix":"","firstName":"Sourish","middleName":"","lastName":"Sarkar","suffix":""},{"id":543066395,"identity":"c47915cf-8d88-4588-8816-57dd9c8ad1b5","order_by":3,"name":"Baishali Ghosh","email":"","orcid":"","institution":"Netaji Subhas Chandra Bose Institute of Pharmacy","correspondingAuthor":false,"prefix":"","firstName":"Baishali","middleName":"","lastName":"Ghosh","suffix":""},{"id":543066396,"identity":"66bf75c5-910a-459e-aba9-9d786b95e052","order_by":4,"name":"Sawan Das","email":"","orcid":"","institution":"Netaji Subhas Chandra Bose Institute of Pharmacy","correspondingAuthor":false,"prefix":"","firstName":"Sawan","middleName":"","lastName":"Das","suffix":""},{"id":543066397,"identity":"105114c1-78d7-4f7d-86d2-8f2ec44f7013","order_by":5,"name":"Rimi Dey","email":"","orcid":"","institution":"Netaji Subhas Chandra Bose Institute of Pharmacy","correspondingAuthor":false,"prefix":"","firstName":"Rimi","middleName":"","lastName":"Dey","suffix":""},{"id":543066398,"identity":"da043959-5ebe-4f43-a7ce-c4f0183b39a7","order_by":6,"name":"Sreejan Manna","email":"","orcid":"","institution":"Brainware University","correspondingAuthor":false,"prefix":"","firstName":"Sreejan","middleName":"","lastName":"Manna","suffix":""},{"id":543066399,"identity":"20f910d3-dfd3-4af3-9c7a-a5e949b23525","order_by":7,"name":"Trishna Das","email":"","orcid":"","institution":"Girijananda Chowdhury University (GCU)","correspondingAuthor":false,"prefix":"","firstName":"Trishna","middleName":"","lastName":"Das","suffix":""}],"badges":[],"createdAt":"2025-10-17 10:23:51","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7885636/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7885636/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s12247-026-10537-7","type":"published","date":"2026-03-10T15:59:23+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":96244197,"identity":"e55a97ce-01e2-45b3-b84c-54f8246d9eb8","added_by":"auto","created_at":"2025-11-19 07:17:56","extension":"jpg","order_by":0,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1269512,"visible":true,"origin":"","legend":"","description":"","filename":"Figure1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/3207c7d7d11738e78afcc2ac.jpg"},{"id":96244158,"identity":"a503fda5-864f-42fb-a537-a3ea30aabe63","added_by":"auto","created_at":"2025-11-19 07:17:53","extension":"doc","order_by":1,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":2725376,"visible":true,"origin":"","legend":"","description":"","filename":"RevisedManuscript1Nanoparticles.doc","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/a546d6baf8878cb1721cd7d8.doc"},{"id":95916901,"identity":"8c51f905-23ba-4573-bf20-01978845e2c9","added_by":"auto","created_at":"2025-11-14 11:57:11","extension":"jpg","order_by":2,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":973800,"visible":true,"origin":"","legend":"","description":"","filename":"Figure10.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/22f07dfb57dc1d618815b2f5.jpg"},{"id":95916903,"identity":"a3c35e6d-65f7-4de6-b7e3-1a300986d620","added_by":"auto","created_at":"2025-11-14 11:57:11","extension":"docx","order_by":3,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":36673,"visible":true,"origin":"","legend":"","description":"","filename":"TableSpringer.docx","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/9d5aed11c6b15169e30cad6a.docx"},{"id":95916900,"identity":"b73ea729-9eee-496b-b235-66cbf497eeca","added_by":"auto","created_at":"2025-11-14 11:57:11","extension":"jpg","order_by":4,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":2147817,"visible":true,"origin":"","legend":"","description":"","filename":"Figure11.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/14f83ef24648fb512ffb662b.jpg"},{"id":95916905,"identity":"a6ea0161-6ac1-4594-ac94-5ef35cd76478","added_by":"auto","created_at":"2025-11-14 11:57:11","extension":"jpg","order_by":5,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1102149,"visible":true,"origin":"","legend":"","description":"","filename":"Figure12.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/c2abccb53d54cfe900981c6f.jpg"},{"id":95916907,"identity":"d8137e78-6272-4ad9-a5d6-15a60e9e2086","added_by":"auto","created_at":"2025-11-14 11:57:11","extension":"jpg","order_by":6,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1407735,"visible":true,"origin":"","legend":"","description":"","filename":"Figure13.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/3a95a8b1a816dafde92482d4.jpg"},{"id":96244782,"identity":"610a5230-d1e1-47b6-a166-c371094828aa","added_by":"auto","created_at":"2025-11-19 07:19:16","extension":"jpg","order_by":7,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1269319,"visible":true,"origin":"","legend":"","description":"","filename":"Figure14.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/b5a32757a681f096f8817a62.jpg"},{"id":96243933,"identity":"196d0234-7fa4-4563-83eb-cb92b9e613db","added_by":"auto","created_at":"2025-11-19 07:17:19","extension":"jpg","order_by":8,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1320989,"visible":true,"origin":"","legend":"","description":"","filename":"Figure15.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/549deb2a0818d421a2ae1366.jpg"},{"id":95916916,"identity":"51a3ede7-6270-41a4-a1e9-054e9a7f9ab5","added_by":"auto","created_at":"2025-11-14 11:57:12","extension":"jpg","order_by":9,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1047060,"visible":true,"origin":"","legend":"","description":"","filename":"Figure16.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/76b7bfa85924164264246c43.jpg"},{"id":96244378,"identity":"58fb86f2-aa65-4716-a802-0e7171d77768","added_by":"auto","created_at":"2025-11-19 07:18:18","extension":"jpg","order_by":10,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1152967,"visible":true,"origin":"","legend":"","description":"","filename":"Figure17.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/d9156388fb25f6ad1a3ebfa8.jpg"},{"id":95916912,"identity":"6587ddc1-f4f7-4264-b2bb-18beedc5b76b","added_by":"auto","created_at":"2025-11-14 11:57:12","extension":"jpg","order_by":11,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":996731,"visible":true,"origin":"","legend":"","description":"","filename":"Figure18.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/e9f180bbd9c648cb1ad4c63e.jpg"},{"id":96242842,"identity":"e6a61c11-9974-472e-8d44-f9646b28b38c","added_by":"auto","created_at":"2025-11-19 07:14:34","extension":"jpg","order_by":12,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1288757,"visible":true,"origin":"","legend":"","description":"","filename":"Figure19.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/189e60db93ba1684ab6b11ec.jpg"},{"id":96244995,"identity":"5d8b7952-d9f5-46fd-b1b1-cc1be57c3552","added_by":"auto","created_at":"2025-11-19 07:19:41","extension":"jpg","order_by":13,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1163513,"visible":true,"origin":"","legend":"","description":"","filename":"Figure2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/afd83b395b86528dc1892dc5.jpg"},{"id":96362818,"identity":"05933ca6-79f0-4e57-a578-9b5b7ae7b2b3","added_by":"auto","created_at":"2025-11-20 09:57:23","extension":"jpg","order_by":14,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":2444168,"visible":true,"origin":"","legend":"","description":"","filename":"Figure20.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/b494e70304f14bca7d049380.jpg"},{"id":95916914,"identity":"4d0a5c44-459d-427d-97f6-505cf3f144e0","added_by":"auto","created_at":"2025-11-14 11:57:12","extension":"jpg","order_by":15,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1526504,"visible":true,"origin":"","legend":"","description":"","filename":"Figure3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/e18cab6e0ce66c00eb350bdb.jpg"},{"id":95916909,"identity":"728bf991-c738-41b0-97bc-482d1dfa9096","added_by":"auto","created_at":"2025-11-14 11:57:12","extension":"jpg","order_by":16,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1336811,"visible":true,"origin":"","legend":"","description":"","filename":"Figure4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/9b7153f0ed1a475596ac66f0.jpg"},{"id":96244090,"identity":"79671279-0710-4b11-85f8-7b9bd8bda9a6","added_by":"auto","created_at":"2025-11-19 07:17:41","extension":"jpg","order_by":17,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1057669,"visible":true,"origin":"","legend":"","description":"","filename":"Figure5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/2822f8ebd746f8624decc2a5.jpg"},{"id":95916942,"identity":"42fe2142-951e-4a67-b040-7d82bb93b66e","added_by":"auto","created_at":"2025-11-14 11:57:12","extension":"jpg","order_by":18,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1362642,"visible":true,"origin":"","legend":"","description":"","filename":"Figure6.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/f093a24fc02997718afd8251.jpg"},{"id":95916932,"identity":"fe6bb600-37bc-4b7b-9c7c-70a7798de954","added_by":"auto","created_at":"2025-11-14 11:57:12","extension":"jpg","order_by":19,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":963941,"visible":true,"origin":"","legend":"","description":"","filename":"Figure7.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/59d2dda891c66a1e644b687b.jpg"},{"id":95916927,"identity":"36aaa879-f42f-4418-bf6a-08d100936997","added_by":"auto","created_at":"2025-11-14 11:57:12","extension":"jpg","order_by":20,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1121354,"visible":true,"origin":"","legend":"","description":"","filename":"Figure8.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/46d710e0f55ccd5af919113e.jpg"},{"id":96242968,"identity":"9d66f0ed-29a1-42f4-9dc7-aa92e4c2ad9a","added_by":"auto","created_at":"2025-11-19 07:15:04","extension":"jpg","order_by":21,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":963342,"visible":true,"origin":"","legend":"","description":"","filename":"Figure9.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/48398a67df5a70efd6592c08.jpg"},{"id":95916898,"identity":"cdb2c7ed-df45-46c3-8ec2-a8b083fff258","added_by":"auto","created_at":"2025-11-14 11:57:11","extension":"json","order_by":22,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":9069,"visible":true,"origin":"","legend":"","description":"","filename":"8a40e985550a4905a416054edf10bccc.json","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/6a4e0ec57452ddf7c06991c9.json"},{"id":96243140,"identity":"679d8cfa-98bd-40e2-bf22-52cc0928c767","added_by":"auto","created_at":"2025-11-19 07:15:41","extension":"jpg","order_by":23,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":3244314,"visible":true,"origin":"","legend":"","description":"","filename":"Graphicalabstract.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/48546e331be8cef8c8298787.jpg"},{"id":95916926,"identity":"d1a2ceac-59ce-4a98-b491-e8c7b1237af2","added_by":"auto","created_at":"2025-11-14 11:57:12","extension":"xml","order_by":24,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":267435,"visible":true,"origin":"","legend":"","description":"","filename":"8a40e985550a4905a416054edf10bccc1enriched.xml","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/13ebe45ada4bc134fb5ea0d2.xml"},{"id":96244434,"identity":"4d576e6a-0c8e-428c-a1c7-83228392dfe7","added_by":"auto","created_at":"2025-11-19 07:18:28","extension":"jpg","order_by":25,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1269512,"visible":true,"origin":"","legend":"","description":"","filename":"Figure1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/cf02ead28be5f36ada0ded3f.jpg"},{"id":96243152,"identity":"707cb85a-2626-481c-bebb-14a1183052bb","added_by":"auto","created_at":"2025-11-19 07:15:45","extension":"jpg","order_by":26,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":973800,"visible":true,"origin":"","legend":"","description":"","filename":"Figure10.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/abdda71cb31356b52d5ef192.jpg"},{"id":95916937,"identity":"36965d2d-e1de-4a68-bfff-a82656b0a184","added_by":"auto","created_at":"2025-11-14 11:57:12","extension":"jpg","order_by":27,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":2147817,"visible":true,"origin":"","legend":"","description":"","filename":"Figure11.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/8cbaa2bfb6ce7adf992c184b.jpg"},{"id":96244246,"identity":"d3bbc7ab-c257-4fa0-95e2-85adff16ce6b","added_by":"auto","created_at":"2025-11-19 07:18:00","extension":"jpg","order_by":28,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1102149,"visible":true,"origin":"","legend":"","description":"","filename":"Figure12.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/649f17f8ea01c7b75e26629b.jpg"},{"id":95916917,"identity":"c67ada5c-2674-4d29-a6ba-a2d891efb6f8","added_by":"auto","created_at":"2025-11-14 11:57:12","extension":"jpg","order_by":29,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1407735,"visible":true,"origin":"","legend":"","description":"","filename":"Figure13.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/6e59dc1428adcaca40cb6b89.jpg"},{"id":96243124,"identity":"9b58933b-33eb-4c58-9be9-e931ae6060d5","added_by":"auto","created_at":"2025-11-19 07:15:36","extension":"jpg","order_by":30,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1269319,"visible":true,"origin":"","legend":"","description":"","filename":"Figure14.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/29b4112e6cad7395de675fe3.jpg"},{"id":96243741,"identity":"737fa427-582b-475d-92d0-51f6e7ce1c15","added_by":"auto","created_at":"2025-11-19 07:16:56","extension":"jpg","order_by":31,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1320989,"visible":true,"origin":"","legend":"","description":"","filename":"Figure15.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/0bcda215b892ac1dccd2af06.jpg"},{"id":95916921,"identity":"6c3ff82e-1048-4f95-a70f-085c991a8d20","added_by":"auto","created_at":"2025-11-14 11:57:12","extension":"jpg","order_by":32,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1047060,"visible":true,"origin":"","legend":"","description":"","filename":"Figure16.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/8f53e61e907d1b4d0fd23361.jpg"},{"id":95916944,"identity":"2619e6ae-8700-42b9-a5c4-0e9ce4a8c925","added_by":"auto","created_at":"2025-11-14 11:57:13","extension":"jpg","order_by":33,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1152967,"visible":true,"origin":"","legend":"","description":"","filename":"Figure17.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/bb134928771c09c13c4aecbd.jpg"},{"id":96243767,"identity":"d27fc9ca-6d95-4586-b05f-181c1882aac3","added_by":"auto","created_at":"2025-11-19 07:17:00","extension":"jpg","order_by":34,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":996731,"visible":true,"origin":"","legend":"","description":"","filename":"Figure18.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/56763a4a1323f88959c8636d.jpg"},{"id":96243759,"identity":"d5a1f148-11a3-4fe5-8438-db0a59a1e42f","added_by":"auto","created_at":"2025-11-19 07:16:59","extension":"jpg","order_by":35,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1288757,"visible":true,"origin":"","legend":"","description":"","filename":"Figure19.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/13463ca0b72c4f7abf02318c.jpg"},{"id":95916948,"identity":"35309af0-d0b1-4634-a701-b50fb732c605","added_by":"auto","created_at":"2025-11-14 11:57:13","extension":"jpg","order_by":36,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1163513,"visible":true,"origin":"","legend":"","description":"","filename":"Figure2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/1e40f691477632ac3471e97e.jpg"},{"id":96243082,"identity":"0bb17e77-2eb5-49f1-988f-9cd0f738e232","added_by":"auto","created_at":"2025-11-19 07:15:29","extension":"jpg","order_by":37,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":2444168,"visible":true,"origin":"","legend":"","description":"","filename":"Figure20.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/eba7f73e2217620fea35ab9a.jpg"},{"id":95916935,"identity":"d504fdb1-54f0-4d47-899c-cae94ec96f72","added_by":"auto","created_at":"2025-11-14 11:57:12","extension":"jpg","order_by":38,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1526504,"visible":true,"origin":"","legend":"","description":"","filename":"Figure3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/d5f25f642222008ee234e123.jpg"},{"id":96243595,"identity":"e27710e4-2ef7-41ca-866d-59ac185f7f13","added_by":"auto","created_at":"2025-11-19 07:16:41","extension":"jpg","order_by":39,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1336811,"visible":true,"origin":"","legend":"","description":"","filename":"Figure4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/3a0ce09f915ae05ea9ff0912.jpg"},{"id":96243390,"identity":"5ec4fd14-237e-4f22-8bca-35c22f1d308d","added_by":"auto","created_at":"2025-11-19 07:16:15","extension":"jpg","order_by":40,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1057669,"visible":true,"origin":"","legend":"","description":"","filename":"Figure5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/4be9ea23d9e4cda80303c928.jpg"},{"id":95916957,"identity":"61d59cbf-81b1-45a6-be81-dac2497b8007","added_by":"auto","created_at":"2025-11-14 11:57:13","extension":"jpg","order_by":41,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1362642,"visible":true,"origin":"","legend":"","description":"","filename":"Figure6.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/52a19e7e66172e80af4bc9a9.jpg"},{"id":95916922,"identity":"b90487b6-4e8c-40a7-b2bb-657e2bc6cde0","added_by":"auto","created_at":"2025-11-14 11:57:12","extension":"jpg","order_by":42,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":963941,"visible":true,"origin":"","legend":"","description":"","filename":"Figure7.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/c030e27741028843c032ecc1.jpg"},{"id":95916923,"identity":"8813e95e-d2b7-42f2-89d4-25f3d1155be4","added_by":"auto","created_at":"2025-11-14 11:57:12","extension":"jpg","order_by":43,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1121354,"visible":true,"origin":"","legend":"","description":"","filename":"Figure8.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/0e04bc6fde851ae3d50b8f10.jpg"},{"id":95916946,"identity":"ee6fb98e-d5f9-4703-9b1e-cb6cabb66d52","added_by":"auto","created_at":"2025-11-14 11:57:13","extension":"jpg","order_by":44,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":963342,"visible":true,"origin":"","legend":"","description":"","filename":"Figure9.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/48fea9e35f2fadd9b8b8a004.jpg"},{"id":95916918,"identity":"84b56f98-f193-43d5-9034-1f241085372e","added_by":"auto","created_at":"2025-11-14 11:57:12","extension":"jpeg","order_by":45,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":292012,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/f45976a2bea48159f0d735ec.jpeg"},{"id":96244128,"identity":"d8383998-b169-4c7d-9ab5-8b5b4e3e7b47","added_by":"auto","created_at":"2025-11-19 07:17:48","extension":"jpeg","order_by":46,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":78824,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage10.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/da09baf96551b76909be4483.jpeg"},{"id":95916956,"identity":"5c89c900-87e5-4706-8b23-0063a3f892cf","added_by":"auto","created_at":"2025-11-14 11:57:13","extension":"jpeg","order_by":47,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":60152,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage11.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/8b9b8929f27d101b9b3bbc05.jpeg"},{"id":96243385,"identity":"7e9b2003-bb5b-4084-b843-eb63711e1ed8","added_by":"auto","created_at":"2025-11-19 07:16:14","extension":"jpeg","order_by":48,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":188328,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage12.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/cd3976130d740562f5fb7047.jpeg"},{"id":95916960,"identity":"96e8e8a2-4cf8-44be-89e7-ecb751fe6505","added_by":"auto","created_at":"2025-11-14 11:57:13","extension":"jpeg","order_by":49,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":98261,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage13.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/a38df7cc93e3a80c7b91997e.jpeg"},{"id":95916929,"identity":"eb5afeb8-5fbe-4109-8153-1d37d8ccc8b3","added_by":"auto","created_at":"2025-11-14 11:57:12","extension":"jpeg","order_by":50,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":114072,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage14.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/afd95d672331ae8418476148.jpeg"},{"id":95916941,"identity":"b9941bb9-c4e1-4276-8f7b-0bacd07c1fad","added_by":"auto","created_at":"2025-11-14 11:57:12","extension":"jpeg","order_by":51,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":140559,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage15.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/8c55818b76ae698521dc2c65.jpeg"},{"id":95916987,"identity":"667fbef3-ec82-478d-9a4b-a170ea8c4c78","added_by":"auto","created_at":"2025-11-14 11:57:14","extension":"jpeg","order_by":52,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":124861,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage16.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/24fbf456ddb0fb42be244b1d.jpeg"},{"id":96243820,"identity":"40fcc107-41a9-4e02-b9af-07d5bf0d2684","added_by":"auto","created_at":"2025-11-19 07:17:06","extension":"jpeg","order_by":53,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":71247,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage17.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/1ff49a501a2c1fe9928a93b0.jpeg"},{"id":96243629,"identity":"e39baf28-dbe7-4395-a825-415ae0045530","added_by":"auto","created_at":"2025-11-19 07:16:46","extension":"jpeg","order_by":54,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":91796,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage18.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/95a0f90914b27ccc6a495d6c.jpeg"},{"id":96243203,"identity":"26f175c0-3f6c-4830-881d-1609c7cb40e7","added_by":"auto","created_at":"2025-11-19 07:15:50","extension":"jpeg","order_by":55,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":96093,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage19.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/5a634a6b775e819cccf0af23.jpeg"},{"id":95916933,"identity":"4fdb45e3-0b9d-45cd-b9b1-4b0dad47175d","added_by":"auto","created_at":"2025-11-14 11:57:12","extension":"jpeg","order_by":56,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":59554,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/55938b2018082d8b1f5ea8ff.jpeg"},{"id":95916985,"identity":"c732185d-2185-4a7d-8c87-2f15449d0519","added_by":"auto","created_at":"2025-11-14 11:57:14","extension":"jpeg","order_by":57,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":113042,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage20.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/19de7b066b4c467926a37c33.jpeg"},{"id":95916938,"identity":"f2ae06f9-12bc-4623-8a1e-64c85832e724","added_by":"auto","created_at":"2025-11-14 11:57:12","extension":"jpeg","order_by":58,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":220440,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage21.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/770520232cda16250988b6be.jpeg"},{"id":95916953,"identity":"054d2e9d-e128-4182-ad3e-485d90dd3748","added_by":"auto","created_at":"2025-11-14 11:57:13","extension":"jpeg","order_by":59,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":81014,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/9d0bdf5c65b962ebb2f15747.jpeg"},{"id":96243542,"identity":"0dcb5586-b850-41ff-a083-79778bfcd79c","added_by":"auto","created_at":"2025-11-19 07:16:39","extension":"jpeg","order_by":60,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":135365,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage4.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/2f563009c32e5f10e774ea81.jpeg"},{"id":95916943,"identity":"1eedb31a-444b-461b-875b-d84fba39427b","added_by":"auto","created_at":"2025-11-14 11:57:13","extension":"jpeg","order_by":61,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":331873,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage5.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/3e353281e3526f5ea6510f56.jpeg"},{"id":96243569,"identity":"7155fab0-ce90-4f8c-89c1-3662724a0c20","added_by":"auto","created_at":"2025-11-19 07:16:40","extension":"jpeg","order_by":62,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":74270,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage6.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/7c60310eb16c0eae069e053e.jpeg"},{"id":96243473,"identity":"722caf22-f33d-4fa1-ad74-ca234cdd5ee4","added_by":"auto","created_at":"2025-11-19 07:16:28","extension":"jpeg","order_by":63,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":330355,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage7.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/dc58363cd61d5b669cbd5855.jpeg"},{"id":95916971,"identity":"62c2ac88-c678-4373-9d37-357d73b1b8c2","added_by":"auto","created_at":"2025-11-14 11:57:13","extension":"jpeg","order_by":64,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":46748,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage8.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/95e7738b050240e954974659.jpeg"},{"id":96243392,"identity":"6e950168-f591-4fb6-944b-0b94d7268903","added_by":"auto","created_at":"2025-11-19 07:16:15","extension":"jpeg","order_by":65,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":82474,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage9.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/e09cf0eda306809aeed2b537.jpeg"},{"id":95916959,"identity":"ce57e0cd-7bff-47af-bd3f-fb9b85caa1c2","added_by":"auto","created_at":"2025-11-14 11:57:13","extension":"png","order_by":66,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":502252,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure1.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/ab966bbd8b02535111a7a836.png"},{"id":96244135,"identity":"cf971496-39b2-45c7-ace1-79f42f8c453b","added_by":"auto","created_at":"2025-11-19 07:17:50","extension":"png","order_by":67,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":220012,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure10.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/3827dca05fb901d75bc6d0e0.png"},{"id":95916983,"identity":"c0d24dcf-0cc5-411b-ad66-42f71cd8b67b","added_by":"auto","created_at":"2025-11-14 11:57:14","extension":"png","order_by":68,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":10133690,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure11.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/d28e60c6cad915bc4e3f695c.png"},{"id":96243905,"identity":"b3fd23b1-d0d3-44ee-8a11-219400c395ea","added_by":"auto","created_at":"2025-11-19 07:17:17","extension":"png","order_by":69,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":360917,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure12.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/5a2800d5e51caceeeb987d5c.png"},{"id":96243558,"identity":"5915d124-3fca-403c-8a47-b6b6511b1452","added_by":"auto","created_at":"2025-11-19 07:16:40","extension":"png","order_by":70,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":694683,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure13.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/cd6b690611103947cfcfbc54.png"},{"id":96243381,"identity":"5fdc8cae-9360-4fb9-8b3d-53da78f80fc2","added_by":"auto","created_at":"2025-11-19 07:16:13","extension":"png","order_by":71,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":454460,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure14.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/80dc01f3b03a0406b0e01f23.png"},{"id":96243610,"identity":"95ecc8dc-ddc4-4095-9310-39ad462d54a8","added_by":"auto","created_at":"2025-11-19 07:16:43","extension":"png","order_by":72,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":436799,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure15.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/a090e8a7dab42e35d1acbb11.png"},{"id":96244406,"identity":"e80d4b30-a0ce-4ff4-95c1-95b044374d28","added_by":"auto","created_at":"2025-11-19 07:18:23","extension":"png","order_by":73,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":425001,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure16.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/583b6884635744b770dee0c6.png"},{"id":96243786,"identity":"226e2d0c-6f27-4d44-9093-bda7af0aa694","added_by":"auto","created_at":"2025-11-19 07:17:02","extension":"png","order_by":74,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":448572,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure17.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/cfed3cf81ddbcb58fffd4176.png"},{"id":96244896,"identity":"f6c0c517-24b2-4016-909d-129ce076e2e7","added_by":"auto","created_at":"2025-11-19 07:19:31","extension":"png","order_by":75,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":310226,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure18.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/a13002c356641470bec2d913.png"},{"id":95916975,"identity":"528ba36f-47de-4f15-af33-33328dfcdff0","added_by":"auto","created_at":"2025-11-14 11:57:13","extension":"png","order_by":76,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":585197,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure19.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/57c3b99d3c3c836e482b2f88.png"},{"id":95916982,"identity":"7cf5533b-dfa0-4104-8671-042448949de5","added_by":"auto","created_at":"2025-11-14 11:57:14","extension":"png","order_by":77,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":452635,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure2.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/e859b306a178c4a418cca9de.png"},{"id":96244919,"identity":"1fc6c1b1-1769-411d-aff9-4db820af5265","added_by":"auto","created_at":"2025-11-19 07:19:32","extension":"png","order_by":78,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1296912,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure20.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/ca05a95a8385abd19d3f1334.png"},{"id":95916998,"identity":"2865211d-e9bf-4f48-9764-d28fb07ac6c0","added_by":"auto","created_at":"2025-11-14 11:57:14","extension":"png","order_by":79,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":515316,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure3.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/fe1c539e9d1925b35b0e8a74.png"},{"id":96243174,"identity":"9921a3c4-3d43-481e-97c6-5e6d71471152","added_by":"auto","created_at":"2025-11-19 07:15:48","extension":"png","order_by":80,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":552106,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure4.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/3305a6113b4b0e10645b7859.png"},{"id":96244415,"identity":"982aea6b-a6b5-4336-ab83-b86929ea8bd3","added_by":"auto","created_at":"2025-11-19 07:18:24","extension":"png","order_by":81,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":385900,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure5.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/4297bf5573308ebe3461c034.png"},{"id":96242871,"identity":"9f2128d2-585f-4fe3-8d6e-5b26e3380f8f","added_by":"auto","created_at":"2025-11-19 07:14:44","extension":"png","order_by":82,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":355863,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure6.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/0006dbf03b697afdee6caf77.png"},{"id":96243249,"identity":"1063544f-9bcb-46cf-9929-24c7c2efccb7","added_by":"auto","created_at":"2025-11-19 07:15:54","extension":"png","order_by":83,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":376117,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure7.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/a7738526a535af0f96ee3678.png"},{"id":96244704,"identity":"1d46b631-b972-482b-a2c2-762863aca68a","added_by":"auto","created_at":"2025-11-19 07:19:06","extension":"png","order_by":84,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":267732,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure8.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/b2ae481349a76f36673c16e7.png"},{"id":95917000,"identity":"998048a6-ffd0-4b5a-83f9-8b5f332bc43f","added_by":"auto","created_at":"2025-11-14 11:57:14","extension":"png","order_by":85,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":324179,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFigure9.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/4f661d39257ab6ad7a6a2792.png"},{"id":95916945,"identity":"739e2547-a073-403c-8e77-0a3c55c788e6","added_by":"auto","created_at":"2025-11-14 11:57:13","extension":"png","order_by":86,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":78488,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/e0b6ba9821154e01c5725176.png"},{"id":96243586,"identity":"f12433f6-ae79-40ae-a62d-e77f625f32c3","added_by":"auto","created_at":"2025-11-19 07:16:41","extension":"png","order_by":87,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":18148,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage10.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/759376fcadb369a2cb92b69f.png"},{"id":95916961,"identity":"7e785488-a67c-463f-a571-77496c6b35d1","added_by":"auto","created_at":"2025-11-14 11:57:13","extension":"png","order_by":88,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":9483,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage11.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/ee07f0f289f93f1a599b0ec1.png"},{"id":95916991,"identity":"fcde3678-59be-42ee-bc9b-8de854e9b2fd","added_by":"auto","created_at":"2025-11-14 11:57:14","extension":"png","order_by":89,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":99554,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage12.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/54806de8042b4256827de277.png"},{"id":96245123,"identity":"88ff51a1-1b5e-4265-b587-b8a667bd8ad9","added_by":"auto","created_at":"2025-11-19 07:19:53","extension":"png","order_by":90,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":24164,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage13.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/6eff8be92d0e39866eaf2f81.png"},{"id":95916976,"identity":"4fd7da82-41e3-4b90-86b5-cacf3894a159","added_by":"auto","created_at":"2025-11-14 11:57:13","extension":"png","order_by":91,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":43634,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage14.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/e8af0111b1eba1544c6c2a11.png"},{"id":96243639,"identity":"93249672-41fe-4ea6-9653-a6d16710a4d6","added_by":"auto","created_at":"2025-11-19 07:16:46","extension":"png","order_by":92,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":46760,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage15.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/0edbce1f3221bff7252cef8f.png"},{"id":96242798,"identity":"cd1dc0f0-5068-4a17-9886-dfa5abcdc4c1","added_by":"auto","created_at":"2025-11-19 07:14:22","extension":"png","order_by":93,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":37789,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage16.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/803c72a324439955fc87e5d6.png"},{"id":96243744,"identity":"81beec42-6d0f-4624-839f-245914b75ec8","added_by":"auto","created_at":"2025-11-19 07:16:56","extension":"png","order_by":94,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":26987,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage17.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/1cba44397259c1928b73ec46.png"},{"id":95916973,"identity":"30ac34ca-f040-4e22-b020-84ebaa5c6cf1","added_by":"auto","created_at":"2025-11-14 11:57:13","extension":"png","order_by":95,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":30523,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage18.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/0b337f6f56254de216aad30c.png"},{"id":95917005,"identity":"4becbf9b-ee7c-4c3c-8580-82e6871add1d","added_by":"auto","created_at":"2025-11-14 11:57:14","extension":"png","order_by":96,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":32848,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage19.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/c5219d0aacbbcaa7fc080101.png"},{"id":95916988,"identity":"241d4e32-1de5-4681-8763-83cf3dc93e48","added_by":"auto","created_at":"2025-11-14 11:57:14","extension":"png","order_by":97,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":20068,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/b3b50b87734c355ef0508d60.png"},{"id":96243222,"identity":"ae016b89-2888-4c8f-ac44-a71d802820e4","added_by":"auto","created_at":"2025-11-19 07:15:52","extension":"png","order_by":98,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":41405,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage20.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/e9ec9ef9113dc2de320df39d.png"},{"id":95916994,"identity":"d20f29ae-5303-4021-8157-2f7cafa7649b","added_by":"auto","created_at":"2025-11-14 11:57:14","extension":"png","order_by":99,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":68912,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage21.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/b20b0c5025f9c327ce39ebd3.png"},{"id":96243189,"identity":"ae77ae94-75c9-43eb-94f6-8f7a8d7fff56","added_by":"auto","created_at":"2025-11-19 07:15:50","extension":"png","order_by":100,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":27438,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/0a0ac71e300b079d686eed2e.png"},{"id":95916951,"identity":"c2e34220-58d1-491d-9c77-9d913d0a892e","added_by":"auto","created_at":"2025-11-14 11:57:13","extension":"png","order_by":101,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":39599,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/3bfc0be62f5aadab6bd985b9.png"},{"id":95916970,"identity":"a99a7dcd-a195-4f4c-bdc2-e29a01111474","added_by":"auto","created_at":"2025-11-14 11:57:13","extension":"png","order_by":102,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":66934,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/fd80114b52ba75167a6e0fa0.png"},{"id":95916977,"identity":"4e7a1115-b8be-4736-b5ae-d4c8acca0e94","added_by":"auto","created_at":"2025-11-14 11:57:14","extension":"png","order_by":103,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":18745,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage6.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/5869c95ac07461acefc4a9e0.png"},{"id":95916965,"identity":"6dfc9cd0-f130-4c81-a06e-47b50a631c3a","added_by":"auto","created_at":"2025-11-14 11:57:13","extension":"png","order_by":104,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":80506,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage7.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/66b8bb08cf8c11d35d1115ae.png"},{"id":96245133,"identity":"f67ef290-726d-408a-a101-e5d0b2d03dd5","added_by":"auto","created_at":"2025-11-19 07:19:55","extension":"png","order_by":105,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":14652,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage8.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/1ee1431f29b35123ed7010b1.png"},{"id":95916995,"identity":"b39f8b90-ac32-4d16-a00a-a36bf547b1b3","added_by":"auto","created_at":"2025-11-14 11:57:14","extension":"png","order_by":106,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":25026,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage9.png","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/01ae47b23f8a98d8c43086c0.png"},{"id":96243182,"identity":"29450960-781f-44a7-b4ed-ef913fef3ab0","added_by":"auto","created_at":"2025-11-19 07:15:49","extension":"xml","order_by":107,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":266777,"visible":true,"origin":"","legend":"","description":"","filename":"8a40e985550a4905a416054edf10bccc1structuring.xml","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/a3a4ebb782b31a7bf8f71098.xml"},{"id":96243508,"identity":"69954c87-214e-46bb-a6d1-3cf860b81572","added_by":"auto","created_at":"2025-11-19 07:16:34","extension":"html","order_by":108,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":281621,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1/d559a6b2c077e1381e165f37.html"},{"id":104739540,"identity":"5988af0e-c961-4d2c-80c4-52771484d76c","added_by":"auto","created_at":"2026-03-16 16:08:56","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1757386,"visible":true,"origin":"","legend":"","description":"","filename":"RevisedManuscript1Nanoparticles.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7885636/v1_covered_f0c535d5-4695-4ffa-9ea7-08316773ff1e.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Fabrication and characterization of polymeric nanoparticle for efficient and sustained delivery of Acyclovir for antiviral therapy and in-silico study using Pk-Sim software","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":true,"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":"Nanoparticle, STPP, Ionic-Gelation, FT-IR, SEM, PBPK","lastPublishedDoi":"10.21203/rs.3.rs-7885636/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7885636/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003ePurpose\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe aim of this study is to develop the nanoparticles of the weakly water-soluble antiviral medication Acyclovir using biodegradable and biocompatible natural polymer to overcome it’ s poor oral bioavailability and frequent dosing frequency were the goals of this work.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe ionic gelation method was used to prepare acyclovir loaded chitosan nanoparticle using STPP (Sodium tripolyphosphate) as the crosslinking agent. The formulations were characterized by determining particle size, surface morphology, percentage of yield, entrapment efficiency, drug- excipient compatibility (FT-IR), \u003cem\u003ein-vitro\u003c/em\u003e release study and \u003cem\u003ein-silico\u003c/em\u003e study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe FT-IR shows no incompatibility between the drug and the excipients. The analysis of dynamic light scattering indicated that nanoparticles were with it’s nanometric range (within 100 nm), demonstrating moderate polydispersity and SEM shows spherical morphology with smooth surface. The entrapment efficiency showed ⁓98% which indicates that chitosan is a potential carrier for the acyclovir. \u003cem\u003eIn-vitro\u003c/em\u003e release study illustrates sustain drug release over the time period and drug release kinetic best fitted in korsemeyer-peppas model which indicates diffusion-controlled release of the drug. Additionally, \u003cem\u003ein-silico\u003c/em\u003e PBPK model predicted improved plasma profile compared to the conventional oral dosage form.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe findings suggested that the optimized formulation, especially F3, holding promise for sustained and controlled delivery of Acyclovir, potentially reducing dosing frequency and minimizing side effects.\u003c/p\u003e","manuscriptTitle":"Fabrication and characterization of polymeric nanoparticle for efficient and sustained delivery of Acyclovir for antiviral therapy and in-silico study using Pk-Sim software","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-14 11:57:06","doi":"10.21203/rs.3.rs-7885636/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":"f9f29115-2999-4fb1-842e-f38b4d52b94c","owner":[],"postedDate":"November 14th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-03-16T16:04:49+00:00","versionOfRecord":{"articleIdentity":"rs-7885636","link":"https://doi.org/10.1007/s12247-026-10537-7","journal":{"identity":"journal-of-pharmaceutical-innovation","isVorOnly":false,"title":"Journal of Pharmaceutical Innovation"},"publishedOn":"2026-03-10 15:59:23","publishedOnDateReadable":"March 10th, 2026"},"versionCreatedAt":"2025-11-14 11:57:06","video":"","vorDoi":"10.1007/s12247-026-10537-7","vorDoiUrl":"https://doi.org/10.1007/s12247-026-10537-7","workflowStages":[]},"version":"v1","identity":"rs-7885636","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7885636","identity":"rs-7885636","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}