{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T23:52:10Z","timestamp":1781913130208,"version":"3.54.5"},"reference-count":50,"publisher":"Georg Thieme Verlag KG","issue":"07","funder":[{"name":"Doctors, Innovations, Science for Children"},{"DOI":"10.13039\/501100006769","name":"Russian Science Foundation","doi-asserted-by":"crossref","award":["23-44-00082"],"award-info":[{"award-number":["23-44-00082"]}],"id":[{"id":"10.13039\/501100006769","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Thromb Haemost"],"published-print":{"date-parts":[[2026,7]]},"abstract":"<jats:p>Microfluidic assays offer a promising solution for accessing the state of the hemostasis system, testing drugs, and adjusting anticoagulant dosages. However, standardization of such assays is still missing. We aimed to design a robust and readily accessible microfluidic assay, which follows recent scientific and standardization committee (SSC) guidelines and is sensitive to hemostatic abnormalities.<\/jats:p>\n                  <jats:p>We optimized key parameters of the whole blood perfusion system to produce a reliable assay suitable for rapid evaluation of primary hemostasis in patients. The optimized protocol includes anticoagulation with hirudin, 5\u2009minutes of perfusion at a shear rate of 1,000\u2009s\u22121 over the millimeter-wide fibrillar collagen patch at room temperature, and evaluation of thrombus formation using the upright fluorescent microscope.<\/jats:p>\n                  <jats:p>Percentage of the activator area covered by thrombi was a reliable parameter demonstrating reproducible results for a given donor over a time course of months. Analysis of the whole blood from 7 patients with Wiscott\u2013Aldrich syndrome, 34 patients with immune thrombocytopenia (ITP), and 8 patients with X-linked agammaglobulinemia showed a significant decrease in thrombus surface coverage compared to that of healthy individuals. Importantly, the microfluidic assay was able to differentiate between ITP patients with distinct clinical bleeding scores better than platelet counts and cytofluorometric parameters.<\/jats:p>\n                  <jats:p>The developed robust microfluidic assay represents an accessible tool for the assessment of primary haemostasis in patients and is promising for clinical use.<\/jats:p>","DOI":"10.1055\/a-2689-7330","type":"journal-article","created":{"date-parts":[[2025,8,26]],"date-time":"2025-08-26T23:02:23Z","timestamp":1756249343000},"page":"702-713","source":"Crossref","is-referenced-by-count":1,"title":["Impairment of Collagen-Induced Thrombus Formation in Microfluidic Assay Correlates with Bleeding Complications Better Than Cytofluorometric Parameters"],"prefix":"10.1055","volume":"126","author":[{"given":"Evgeniya A.","family":"Melnikova","sequence":"additional","affiliation":[{"name":"Center for Theoretical Problems of Physico-Chemical Pharmacology, Russian Academy of Sciences, Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tatiana A.","family":"Kovalenko","sequence":"additional","affiliation":[{"name":"Center for Theoretical Problems of Physico-Chemical Pharmacology, Russian Academy of Sciences, Moscow, Russia"},{"name":"Dmitry Rogachev National Medical Research Cent\u0435r of Pediatric Hematology, Oncology and Immunology, Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lilia","family":"Pogodina","sequence":"additional","affiliation":[{"name":"Dmitry Rogachev National Medical Research Cent\u0435r of Pediatric Hematology, Oncology and Immunology, Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anastasia A.","family":"Ignatova","sequence":"additional","affiliation":[{"name":"Dmitry Rogachev National Medical Research Cent\u0435r of Pediatric Hematology, Oncology and Immunology, Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elena V.","family":"Suntsova","sequence":"additional","affiliation":[{"name":"Dmitry Rogachev National Medical Research Cent\u0435r of Pediatric Hematology, Oncology and Immunology, Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna Yu.","family":"Shcherbina","sequence":"additional","affiliation":[{"name":"Dmitry Rogachev National Medical Research Cent\u0435r of Pediatric Hematology, Oncology and Immunology, Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Galina A.","family":"Novichkova","sequence":"additional","affiliation":[{"name":"Dmitry Rogachev National Medical Research Cent\u0435r of Pediatric Hematology, Oncology and Immunology, Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mikhail A.","family":"Panteleev","sequence":"additional","affiliation":[{"name":"Center for Theoretical Problems of Physico-Chemical Pharmacology, Russian Academy of Sciences, Moscow, Russia"},{"name":"Dmitry Rogachev National Medical Research Cent\u0435r of Pediatric Hematology, Oncology and Immunology, Moscow, Russia"},{"name":"Faculty of Physics, M.V. Lomonosov Moscow State University, Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dmitry Yu.","family":"Nechipurenko","sequence":"additional","affiliation":[{"name":"Center for Theoretical Problems of Physico-Chemical Pharmacology, Russian Academy of Sciences, Moscow, Russia"},{"name":"Dmitry Rogachev National Medical Research Cent\u0435r of Pediatric Hematology, Oncology and Immunology, Moscow, Russia"},{"name":"Faculty of Physics, M.V. Lomonosov Moscow State University, Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"194","published-online":{"date-parts":[[2025,9,8]]},"reference":[{"issue":"05","key":"ref1","doi-asserted-by":"crossref","first-page":"417","DOI":"10.1097\/MOH.0b013e3283642186","article-title":"The use of microfluidics in hemostasis: clinical diagnostics and biomimetic models of vascular injury","volume":"20","author":"K B Neeves","year":"2013","journal-title":"Curr Opin Hematol"},{"issue":"01","key":"ref2","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1007\/s12195-016-0469-0","article-title":"A microfluidic model of hemostasis sensitive to platelet function and coagulation","volume":"10","author":"R M Schoeman","year":"2017","journal-title":"Cell Mol Bioeng"},{"issue":"01","key":"ref3","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1038\/s41467-018-02990-x","article-title":"A microengineered vascularized bleeding model that integrates the principal components of hemostasis","volume":"9","author":"Y Sakurai","year":"2018","journal-title":"Nat Commun"},{"issue":"01","key":"ref4","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1161\/ATVBAHA.118.311390","article-title":"Clot contraction drives the translocation of procoagulant platelets to thrombus surface","volume":"39","author":"D Y Nechipurenko","year":"2019","journal-title":"Arterioscler Thromb Vasc Biol"},{"issue":"10","key":"ref5","doi-asserted-by":"crossref","first-page":"2471","DOI":"10.3324\/haematol.2019.235754","article-title":"Shear rate gradients promote a bi-phasic thrombus formation on weak adhesive proteins, such as fibrinogen in a VWF-dependent manner","volume":"105","author":"N Receveur","year":"2020","journal-title":"Haematologica"},{"issue":"01","key":"ref6","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1134\/S1990747822020040","article-title":"The role of immobilized phospholipids in the initiation of blood coagulation under flow conditions","volume":"16","author":"A D Megalinskiy","year":"2022","journal-title":"Biochem (Moscow), Suppl Ser A Membr Cell Biol"},{"issue":"03","key":"ref7","doi-asserted-by":"crossref","first-page":"748","DOI":"10.1111\/jth.14717","article-title":"In vitro flow based systems to study platelet function and thrombus formation: Recommendations for standardization: Communication from the SSC on Biorheology of the ISTH","volume":"18","author":"P H Mangin","year":"2020","journal-title":"J Thromb Haemost"},{"issue":"01","key":"ref8","doi-asserted-by":"crossref","first-page":"e82493","DOI":"10.1371\/journal.pone.0082493","article-title":"Microfluidic thrombosis under multiple shear rates and antiplatelet therapy doses","volume":"9","author":"M Li","year":"2014","journal-title":"PLoS ONE"},{"key":"ref9","doi-asserted-by":"crossref","first-page":"779753","DOI":"10.3389\/fphar.2021.779753","article-title":"Platelet mechanobiology inspired microdevices: From hematological function tests to 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Lehmann","year":"2018","journal-title":"J Thromb Haemost"},{"issue":"06","key":"ref16","doi-asserted-by":"crossref","first-page":"1256","DOI":"10.3324\/haematol.2018.198853","article-title":"High-throughput elucidation of thrombus formation reveals sources of platelet function variability","volume":"104","author":"J P van Geffen","year":"2019","journal-title":"Haematologica"},{"issue":"20","key":"ref17","doi-asserted-by":"crossref","first-page":"6163","DOI":"10.1182\/bloodadvances.2023009860","article-title":"High-throughput microfluidic blood testing to phenotype genetically linked platelet disorders: an aid to diagnosis","volume":"7","author":"D I Fernandez","year":"2023","journal-title":"Blood Adv"},{"key":"ref18","doi-asserted-by":"crossref","first-page":"10176","DOI":"10.1038\/ncomms10176","article-title":"A shear gradient-activated microfluidic device for automated monitoring of whole blood haemostasis and platelet function","volume":"7","author":"A Jain","year":"2016","journal-title":"Nat Commun"},{"issue":"01","key":"ref19","doi-asserted-by":"crossref","first-page":"408","DOI":"10.1172\/JCI58753","article-title":"In vitro modeling of the microvascular occlusion and thrombosis that occur in hematologic diseases using microfluidic technology","volume":"122","author":"M Tsai","year":"2012","journal-title":"J Clin Invest"},{"issue":"11","key":"ref20","doi-asserted-by":"crossref","first-page":"1824","DOI":"10.1182\/blood-2014-01-550319","article-title":"A systems approach to hemostasis: 3. 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Mazurov","year":"2002","journal-title":"Platelets"},{"issue":"10","key":"ref29","doi-asserted-by":"crossref","first-page":"2941","DOI":"10.1016\/j.jtha.2023.07.006","article-title":"Consensus report on flow cytometry for platelet function testing in thrombocytopenic patients: communication from the SSC of the ISTH","volume":"21","author":"G Jourdi","year":"2023","journal-title":"J Thromb Haemost"},{"issue":"04","key":"ref30","doi-asserted-by":"crossref","first-page":"428","DOI":"10.1080\/09537104.2018.1513473","article-title":"Flow cytometry for pediatric platelets","volume":"30","author":"A A Ignatova","year":"2019","journal-title":"Platelets"},{"issue":"01","key":"ref31","doi-asserted-by":"crossref","first-page":"9401","DOI":"10.1038\/s41598-021-88900-6","article-title":"Platelet function and bleeding at different phases of childhood immune thrombocytopenia","volume":"11","author":"A A Ignatova","year":"2021","journal-title":"Sci 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Poulter","year":"2015","journal-title":"Nat Commun"},{"issue":"05","key":"ref45","doi-asserted-by":"crossref","first-page":"506","DOI":"10.1038\/sj.embor.7400665","article-title":"Wiskott-Aldrich syndrome protein is involved in alphaIIb beta3-mediated cell adhesion","volume":"7","author":"S Tsuboi","year":"2006","journal-title":"EMBO Rep"},{"issue":"03","key":"ref46","doi-asserted-by":"crossref","first-page":"416","DOI":"10.1111\/j.1365-2141.2009.07959.x","article-title":"WASP plays a novel role in regulating platelet responses dependent on alphaIIbbeta3 integrin outside-in signalling","volume":"148","author":"A Shcherbina","year":"2010","journal-title":"Br J Haematol"},{"issue":"02","key":"ref47","doi-asserted-by":"crossref","first-page":"230","DOI":"10.1038\/nmat4772","article-title":"Single-platelet nanomechanics measured by high-throughput cytometry","volume":"16","author":"D R Myers","year":"2017","journal-title":"Nat 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