A Microfluidic Model of Hemostasis Sensitive to Platelet Function and Coagulation
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Hemostasis is the process of sealing a vascular injury with a thrombus to arrest bleeding. The type of thrombus that forms depends on the nature of the injury and hemodynamics. There are many models of intravascular thrombus formation whereby blood is exposed to prothrombotic molecules on a solid substrate. However, there are few models of extravascular thrombus formation whereby blood escapes into the extravascular space through a hole in the vessel wall. Here, we describe a microfluidic model of hemostasis that includes vascular, vessel wall, and extravascular compartments. Type I collagen and tissue factor, which support platelet adhesion and initiate coagulation, respectively, were adsorbed to the wall of the injury channel and act synergistically to yield a stable thrombus that stops blood loss into the extravascular compartment in ~ 7.5 min. Inhibiting factor VIII to mimic hemophilia A results in an unstable thrombus that was unable to close the injury. Treatment with a P2Y12 antagonist to reduce platelet activation prolonged the closure time two-fold compared to controls. Taken together, these data demonstrate a hemostatic model that is sensitive to both coagulation and platelet function and can be used to study coagulopathies and platelet dysfunction that result in excessive blood loss.
keywordsBiorheology Biotransport Platelets Coagulation
This work was supported by a NSF CAREER (CBET-1351672), American Heart Association (14GRNT20410094), and the National Institutes of Health (R01HL120728, R21NS082933).
No animal studies were carried out by the authors for this article.
Conflict of Interest
The authors R. M. Schoeman, K. Rana, N. Danes, and M. Lehmann declare that they have no conflicts of interest. J. A. Di Paola, A. L. Fogelson, K. Leiderman, and K. B. Neeves reports Grants from NIH and NSF. A. L. Fogelson reports personal fees from Genentech outside the submitted work.
All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (University of Colorado, Boulder) and with the Helsinki Declaration of 1975, as revised in 2000. Informed consent was obtained from all subjects for being included in the study.
- 1.Ayachit, U. The ParaView Guide: A Parallel Visualization Application. New York: Kitware, Inc, 2015.Google Scholar
- 7.de Witt, S. M., F. Swieringa, R. Cavill, M. M. E. Lamers, R. Van Kruchten, T. Mastenbroek, C. Baaten, S. Coort, N. Pugh, A. Schulz, I. Scharrer, K. Jurk, B. Zieger, K. J. Clemetson, R. W. Farndale, J. W. M. Heemskerk, and J. M. E. M. Cosemans. Identification of platelet function defects by multi-parameter assessment of thrombus formation. Nat. Commun. 5:4257, 2014.Google Scholar
- 8.Drake, T. A., J. H. Morrissey, and T. S. Edgington. Selective cellular expression of tissue factor in human tissues. Implications for disorders of hemostasis and thrombosis. Am. J. Pathol. 134:1087, 1989.Google Scholar
- 13.Goldsmith, H. L., and V. T. Turitto. Rheological aspects of thrombosis and haemostasis: basic principles and applications. ICTH-Report-Subcommittee on Rheology of the International Committee on Thrombosis and Haemostasis. Thromb. Haemost. 55:415–435, 1986.Google Scholar
- 16.Jen, C. J., and J. S. Lin. Direct observation of platelet adhesion to fibrinogen- and fibrin-coated surfaces. Am. J. Physiol. 261:H1457–H1463, 1991.Google Scholar
- 21.Mann, K. G., M. E. Nesheim, W. R. Church, P. Haley, and S. Krishnaswamy. Surface-dependent reactions of the vitamin K-dependent enzyme complexes. Blood 76:1–16, 1990.Google Scholar
- 23.McCarty, O. J. T., D. Ku, M. Sugimoto, M. R. King, J. M. E. M. Cosemans, K. B. Neeves. The Subcommittee on Biorheology. Dimensional analysis and scaling relevant to flow models of thrombus formation: communication from the SSC of the ISTH. J. Thromb. Haemost. 14:619–622, 2016.Google Scholar
- 24.Monroe, D. M., and M. Hoffman. Coagulation factor interaction with platelets. Thromb. Haemost. 88:179, 2002.Google Scholar
- 27.Neeves, K. B., A. A. Onasoga, R. R. Hansen, J. J. Lilly, D. Venckunaite, M. B. Sumner, A. T. Irish, G. Brodsky, M. J. Manco-Johnson, and J. A. Di Paola. Sources of Variability in Platelet Accumulation on Type 1 Fibrillar Collagen in Microfluidic Flow Assays. PLoS One 8:e54680, 2013.CrossRefGoogle Scholar
- 28.Neeves, K. B., O. J. T. McCarty, A. J. Reininger, M. Sugimoto, M. R. King. Biorheology Subcommittee of the SSC of the ISTH. Flow-dependent thrombin and fibrin generation in vitro: opportunities for standardization: communication from SSC of the ISTH. J. Thromb. Haemost. 12:418–420, 2014.Google Scholar
- 29.Nichols, W. L., M. B. Hultin, A. H. James, M. J. Manco-Johnson, R. R. Montgomery, T. L. Ortel, M. E. Rick, J. E. Sadler, M. Weinstein, and B. P. Yawn. von Willebrand disease (VWD): evidence-based diagnosis and management guidelines, the National Heart, Lung, and Blood Institute (NHLBI) Expert Panel report (USA). Haemophilia 14:171–232, 2008.CrossRefGoogle Scholar
- 31.Okorie, U. M., W. S. Denney, M. S. Chatterjee, K. B. Neeves, and S. L. Diamond. Determination of surface tissue factor thresholds that trigger coagulation at venous and arterial shear rates: amplification of 100 fM circulating tissue factor requires flow. Blood 111:3507–3513, 2008.CrossRefGoogle Scholar
- 34.Pries, A. R., D. Neuhaus, and P. Gaehtgens. Blood viscosity in tube flow: dependence on diameter and hematocrit. Am. J. Physiol. 263:H1770–H1778, 1992.Google Scholar
- 47.van Gestel, M. A., J. W. M. Heemskerk, D. W. Slaaf, V. V. T. Heijnen, S. O. Sage, R. S. Reneman, and M. G. A. Oude Egbrink. Real-time detection of activation patterns in individual platelets during thromboembolism in vivo: differences between thrombus growth and embolus formation. J. Vasc. Res. 39:534–543, 2002.Google Scholar
- 48.van Gestel, M. A., S. Reitsma, D. W. Slaaf, V. V. T. Heijnen, M. A. H. Feijge, T. Lindhout, M. A. M. J. van Zandvoort, M. Elg, R. S. Reneman, J. W. M. Heemskerk, and M. G. A. Oude Egbrink. Both ADP and thrombin regulate arteriolar thrombus stabilization and embolization, but are not involved in initial hemostasis as induced by micropuncture. Microcirculation 14:193–205, 2007.Google Scholar
- 51.Weisel, J. Fibrinogen and fibrin. Adv. Protein Chem. 70:248–299, 2005.Google Scholar
- 52.Westein, E., A. D. van der Meer, M. J. E. Kuijpers, J.-P. Frimat, A. van den Berg, and J. W. M. Heemskerk. Atherosclerotic geometries exacerbate pathological thrombus formation poststenosis in a von Willebrand factor-dependent manner. Proc. Natl. Acad. Sci. 110:1357–1362, 2013.CrossRefGoogle Scholar