Engineering tough blood clots for rapid hemostasis and enhanced regeneration

Engineering tough blood clots for rapid hemostasis and enhanced regeneration | 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 Physical Sciences - Article Engineering tough blood clots for rapid hemostasis and enhanced regeneration Jianyu Li, Shuaibing Jiang, Guangyu Bao, Zhen Yang, Xingwei Yang, and 8 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3326102/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 29 Apr, 2026 Read the published version in Nature → Version 1 posted You are reading this latest preprint version Abstract Blood clots play a pivotal role for hemostasis and regeneration1, but are slow to form and mechanically weak. These limitations are attributed to complex coagulation cascades, a high content of mechanically ineffective cells, and a low content of structural polymers2. They pose a considerable risk for life-threatening hemorrhage and constrain the broader application of blood clots3-5. Overcoming these limitations is a critical challenge, as existing strategies focused on polymer networks are inapplicable to highly cellularized materials like blood clots. Here we show a strategy termed click clotting, which rapidly crosslinks living cells into tough cytogels through bioorthogonal click reactions. We demonstrate this strategy by directly crosslinking red blood cells into cytogels and incorporating them within blood clots. The resulting engineered blood clots (EBC) exhibit instantaneous clot formation (< 5 seconds), a 13-fold increase in fracture toughness, and a 4-fold improvement in adhesion energy compared to native blood clots. Experiments and computational modeling reveal a unique toughening mechanism based on cell rupture. In vivo studies show that EBC outperform clinically used products in managing non-compressible hemorrhage. Specifically, EBC can rapidly halt hemorrhage, promote tissue regeneration, mitigate inflammation and foreign body reactions, and prevent postoperative adhesion. Furthermore, our strategy is versatile and applicable to a range of cells and polymers. This work is expected to motivate the development and translation of highly cellularized materials for wound management, bleeding control, tissue repair and regeneration. Physical sciences/Materials science/Biomaterials/Biomedical materials Physical sciences/Materials science/Soft materials/Gels and hydrogels Physical sciences/Materials science/Biomaterials/Biomaterials – cells Full Text Additional Declarations Yes there is potential Competing Interest. Shuaibing Jiang, Guangyu Bao and Jianyu Li are inventors of a provisional patent application that covers the design and application of tough cytogels. Supplementary Files RBCcytogel.mp4 Supplementary Video S1 EBChemostasis.mp4 Supplementary Video S1 Cite Share Download PDF Status: Published Journal Publication published 29 Apr, 2026 Read the published version in Nature → 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-3326102","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Physical Sciences - Article","associatedPublications":[],"authors":[{"id":247273650,"identity":"5e2e9126-77ce-418f-9342-3b2b2f4261e2","order_by":0,"name":"Jianyu Li","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA1ElEQVRIiWNgGAWjYBACxmYILcPG3gCkDCxkiNbCw8ZzAKRFgodo23gYJBJANBFamNuZnz3mbWPg4ZN8nfjxSwFQC/vhBwQcxmZuDNLCJp27WVoG5DCeNAMCWhjMpKFatjFLgLRIMBDSwv4NokXyLEwL+wcCWnigtkjwbmP8ANbCQ8gWnjLJOeckgIEM9AsokNl4cgrwajHsP75N4k2ZjZx8+9mNH3/8sZHjZz++Ab+WBgYGJpCXQYAZFClseNUDgTzIcT9grvyBT+koGAWjYBSMWAAAVL4wmv7KWMwAAAAASUVORK5CYII=","orcid":"https://orcid.org/0000-0002-1128-6804","institution":"McGill University","correspondingAuthor":true,"prefix":"","firstName":"Jianyu","middleName":"","lastName":"Li","suffix":""},{"id":247273651,"identity":"1e99de3c-33b7-4841-b95c-c2f9ab1853b1","order_by":1,"name":"Shuaibing Jiang","email":"","orcid":"https://orcid.org/0000-0002-7402-0587","institution":"McGill University","correspondingAuthor":false,"prefix":"","firstName":"Shuaibing","middleName":"","lastName":"Jiang","suffix":""},{"id":247273652,"identity":"e406bfcb-8d4e-426f-a586-6029061947a2","order_by":2,"name":"Guangyu Bao","email":"","orcid":"https://orcid.org/0000-0001-5794-9020","institution":"McGill University","correspondingAuthor":false,"prefix":"","firstName":"Guangyu","middleName":"","lastName":"Bao","suffix":""},{"id":247273653,"identity":"9e39d86f-8c79-425b-a120-49d504e22804","order_by":3,"name":"Zhen Yang","email":"","orcid":"","institution":"McGill University","correspondingAuthor":false,"prefix":"","firstName":"Zhen","middleName":"","lastName":"Yang","suffix":""},{"id":247273654,"identity":"797aacd0-2809-480d-b762-c2b279f81d59","order_by":4,"name":"Xingwei Yang","email":"","orcid":"","institution":"University of Colorado Boulder","correspondingAuthor":false,"prefix":"","firstName":"Xingwei","middleName":"","lastName":"Yang","suffix":""},{"id":247273655,"identity":"040db2cb-18eb-495a-8265-dc1b2c77eea0","order_by":5,"name":"Joo Eun Kim","email":"","orcid":"","institution":"McGill University","correspondingAuthor":false,"prefix":"","firstName":"Joo","middleName":"Eun","lastName":"Kim","suffix":""},{"id":247273656,"identity":"1c060d40-27e9-4e06-828c-d4b378048e34","order_by":6,"name":"Alexander Nottegar","email":"","orcid":"","institution":"McGill University","correspondingAuthor":false,"prefix":"","firstName":"Alexander","middleName":"","lastName":"Nottegar","suffix":""},{"id":247273657,"identity":"1001cb8f-6d5a-4872-87a9-c2435dcdaf24","order_by":7,"name":"Yin Liu","email":"","orcid":"https://orcid.org/0000-0001-7874-2301","institution":"McGill University","correspondingAuthor":false,"prefix":"","firstName":"Yin","middleName":"","lastName":"Liu","suffix":""},{"id":247273658,"identity":"4a280b23-147d-450c-80f7-c61eb5eb8806","order_by":8,"name":"Zu-hua Gao","email":"","orcid":"","institution":"University of British Columbia","correspondingAuthor":false,"prefix":"","firstName":"Zu-hua","middleName":"","lastName":"Gao","suffix":""},{"id":247273659,"identity":"0c8c2633-4334-4c9c-a3b0-955e4b59d331","order_by":9,"name":"Andrew Beckett","email":"","orcid":"","institution":"University of Toronto","correspondingAuthor":false,"prefix":"","firstName":"Andrew","middleName":"","lastName":"Beckett","suffix":""},{"id":247273660,"identity":"8828aaf5-c1fd-443e-856b-90c12df4c8ac","order_by":10,"name":"Anastasia Nijnik","email":"","orcid":"https://orcid.org/0000-0001-6048-4631","institution":"McGill University","correspondingAuthor":false,"prefix":"","firstName":"Anastasia","middleName":"","lastName":"Nijnik","suffix":""},{"id":247273661,"identity":"f0956a91-ac94-4939-bc1b-a87957d725fa","order_by":11,"name":"Rong Long","email":"","orcid":"https://orcid.org/0000-0001-8992-2420","institution":"University of Colorado Boulder","correspondingAuthor":false,"prefix":"","firstName":"Rong","middleName":"","lastName":"Long","suffix":""},{"id":247273662,"identity":"5b2445cf-6d7e-4943-be1e-ecb9a968c4dc","order_by":12,"name":"Christian Kastrup","email":"","orcid":"","institution":"University of British Columbia","correspondingAuthor":false,"prefix":"","firstName":"Christian","middleName":"","lastName":"Kastrup","suffix":""}],"badges":[],"createdAt":"2023-09-05 03:50:28","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3326102/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3326102/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41586-026-10412-y","type":"published","date":"2026-04-29T04:00:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":108978505,"identity":"05325b36-ff53-4fe3-9b88-53e078a2b3f0","added_by":"auto","created_at":"2026-05-11 11:41:29","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":8964661,"visible":true,"origin":"","legend":"Article File","description":"","filename":"EBCupdatedmanuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3326102/v1_covered_513aae1f-f77b-4a38-ade2-1884ce05f8ea.pdf"},{"id":80826646,"identity":"62ed84cf-1322-4bbe-8767-dff89bc08770","added_by":"auto","created_at":"2025-04-17 13:22:05","extension":"mp4","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":3276800,"visible":true,"origin":"","legend":"Supplementary Video S1","description":"","filename":"RBCcytogel.mp4","url":"https://assets-eu.researchsquare.com/files/rs-3326102/v1/98b23282d304a90c20757a49.mp4"},{"id":80826647,"identity":"db814f09-3d3e-477a-a21a-b58f2459cedb","added_by":"auto","created_at":"2025-04-17 13:22:05","extension":"mp4","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":7491452,"visible":true,"origin":"","legend":"Supplementary Video S1","description":"","filename":"EBChemostasis.mp4","url":"https://assets-eu.researchsquare.com/files/rs-3326102/v1/8de421d27cb915b4131015d5.mp4"}],"financialInterests":"\u003cb\u003eYes\u003c/b\u003e there is potential Competing Interest.\nShuaibing Jiang, Guangyu Bao and Jianyu Li are inventors of a provisional patent application that covers the design and application of tough cytogels.","formattedTitle":"Engineering tough blood clots for rapid hemostasis and enhanced regeneration","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"nature-portfolio","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"","title":"Nature Portfolio","twitterHandle":"","acdcEnabled":false,"dfaEnabled":false,"editorialSystem":"ejp","reportingPortfolio":"","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-3326102/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3326102/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Blood clots play a pivotal role for hemostasis and regeneration1, but are slow to form and mechanically weak. These limitations are attributed to complex coagulation cascades, a high content of mechanically ineffective cells, and a low content of structural polymers2. They pose a considerable risk for life-threatening hemorrhage and constrain the broader application of blood clots3-5. Overcoming these limitations is a critical challenge, as existing strategies focused on polymer networks are inapplicable to highly cellularized materials like blood clots. Here we show a strategy termed click clotting, which rapidly crosslinks living cells into tough cytogels through bioorthogonal click reactions. We demonstrate this strategy by directly crosslinking red blood cells into cytogels and incorporating them within blood clots. The resulting engineered blood clots (EBC) exhibit instantaneous clot formation (\u003c 5 seconds), a 13-fold increase in fracture toughness, and a 4-fold improvement in adhesion energy compared to native blood clots. Experiments and computational modeling reveal a unique toughening mechanism based on cell rupture. In vivo studies show that EBC outperform clinically used products in managing non-compressible hemorrhage. Specifically, EBC can rapidly halt hemorrhage, promote tissue regeneration, mitigate inflammation and foreign body reactions, and prevent postoperative adhesion. Furthermore, our strategy is versatile and applicable to a range of cells and polymers. This work is expected to motivate the development and translation of highly cellularized materials for wound management, bleeding control, tissue repair and regeneration.","manuscriptTitle":"Engineering tough blood clots for rapid hemostasis and enhanced regeneration","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-04-17 13:22:00","doi":"10.21203/rs.3.rs-3326102/v1","editorialEvents":[],"status":"published","journal":{"display":false,"email":"[email protected]","identity":"nature","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"nature","sideBox":"Learn more about [Nature](http://www.nature.com/nature/)","snPcode":"","submissionUrl":"","title":"Nature","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"bedfb489-0ba6-4419-956d-2ba3d8de2293","owner":[],"postedDate":"April 17th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":26110729,"name":"Physical sciences/Materials science/Biomaterials/Biomedical materials"},{"id":26110730,"name":"Physical sciences/Materials science/Soft materials/Gels and hydrogels"},{"id":26110731,"name":"Physical sciences/Materials science/Biomaterials/Biomaterials \u0026#x2013; cells"}],"tags":[],"updatedAt":"2026-05-05T09:41:24+00:00","versionOfRecord":{"articleIdentity":"rs-3326102","link":"https://doi.org/10.1038/s41586-026-10412-y","journal":{"identity":"nature","isVorOnly":false,"title":"Nature"},"publishedOn":"2026-04-29 04:00:00","publishedOnDateReadable":"April 29th, 2026"},"versionCreatedAt":"2025-04-17 13:22:00","video":"","vorDoi":"10.1038/s41586-026-10412-y","vorDoiUrl":"https://doi.org/10.1038/s41586-026-10412-y","workflowStages":[]},"version":"v1","identity":"rs-3326102","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3326102","identity":"rs-3326102","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}