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Dynamics of Blood Flow and Thrombus Formation in a Multi-Bypass Microfluidic Ladder Network

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Abstract

The reaction dynamics of a complex mixture of cells and proteins, such as blood, in branched circulatory networks within the human microvasculature or extravascular therapeutic devices such as extracorporeal oxygenation machine (ECMO) remains ill-defined. In this report we utilize a multi-bypass microfluidics ladder network design with dimensions mimicking venules to study patterns of blood platelet aggregation and fibrin formation under complex shear. Complex blood fluid dynamics within multi-bypass networks under flow were modeled using COMSOL. Red blood cells and platelets were assumed to be non-interacting spherical particles transported by the bulk fluid flow, and convection of the activated coagulation factor II, thrombin, was assumed to be governed by mass transfer. This model served as the basis for predicting formation of local shear rate gradients, stagnation points and recirculation zones as dictated by the bypass geometry. Based on the insights from these models, we were able to predict the patterns of blood clot formation at specific locations in the device. Our experimental data was then used to adjust the model to account for the dynamical presence of thrombus formation in the biorheology of blood flow. The model predictions were then compared to results from experiments using recalcified whole human blood. Microfluidic devices were coated with the extracellular matrix protein, fibrillar collagen, and the initiator of the extrinsic pathway of coagulation, tissue factor. Blood was perfused through the devices at a flow rate of 2 µL/min, translating to physiologically relevant initial shear rates of 300 and 700 s−1 for main channels and bypasses, respectively. Using fluorescent and light microscopy, we observed distinct flow and thrombus formation patterns near channel intersections at bypass points, within recirculation zones and at stagnation points. Findings from this proof-of-principle ladder network model suggest a specific correlation between microvascular geometry and thrombus formation dynamics under shear. This model holds potential for use as an integrative approach to identify regions susceptible to intravascular thrombus formation within the microvasculature as well as extravascular devices such as ECMO.

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Abbreviations

PDMS:

Polydimethylsiloxane

RBCs:

Red blood cells

TF:

Tissue factor

VWF:

Von Willebrand factor

DiOC6 :

3,3′-Dihexyloxacarbocyanine iodide

References

  1. Aird, W. C. Vascular bed-specific thrombosis. J. Thromb. Haemost. 5:283–291, 2007.

    Article  Google Scholar 

  2. Baker-Groberg, S. M., F. A. Cianchetti, K. G. Phillips, and O. J. T. McCarty. Development of a method to quantify platelet adhesion and aggregation under static conditions. Cell. Mol. Bioeng. 7:285–290, 2014.

    Article  Google Scholar 

  3. Baker-Groberg, S. M., S. Lattimore, M. Recht, O. J. T. McCarty, and K. M. Haley. Assessment of neonatal platelet adhesion, activation, and aggregation. J. Thromb. Haemost. 14:815–827, 2016.

    Article  Google Scholar 

  4. Bark, D. L., and D. N. Ku. Platelet transport rates and binding kinetics at high shear over a thrombus. Biophys. J. 105:502–511, 2013.

    Article  Google Scholar 

  5. Bird, R. B., W. E. Stewart, and E. N. Lightfoot. Transport Phenomena. New York: Wiley, 1960.

    Google Scholar 

  6. Boyer, C. J., and R. D. Swartz. Severe clotting during extracorporeal dialysis procedures. Sem. Dial. 4:69–71, 1991.

    Article  Google Scholar 

  7. Brækkan, S. K., E. B. Mathiesen, I. Njølstad, T. Wilsgaard, and J.-B. Hansen. Hematocrit and risk of venous thromboembolism in a general population. The Tromsø study. Haematologica 95:270–275, 2010.

    Article  Google Scholar 

  8. Burgin, T. D., H. Johnson, A. Chung, and J. McGrath. Analytical and finite element modeling of nanomembranes for miniaturized, continuous hemodialysis. Membranes (Basel) 6(1):6, 2015.

    Article  Google Scholar 

  9. Carson, L., and V. M. Doctor. Mechanism of potentiation of antithrombin III and heparin cofactor II inhibition by sulfated xylans. Thromb. Res. 58:367–381, 1990.

    Article  Google Scholar 

  10. Chang, J. Y. Thrombin specificity. Requirement for apolar amino acids adjacent to the thrombin cleavage site of polypeptide substrate. Eur. J. Biochem. 151:217–224, 1985.

    Article  Google Scholar 

  11. Chiu, W.-C., M. J. Slepian, and D. Bluestein. Thrombus formation patterns in the HeartMate II ventricular assist device: clinical observations can be predicted by numerical simulations. ASAIO J. 60:237–240, 2014.

    Article  Google Scholar 

  12. Colace, T. V., G. W. Tormoen, O. J. T. McCarty, and S. L. Diamond. Microfluidics and coagulation biology. Annu. Rev. Biomed. Eng. 15:283–303, 2013.

    Article  Google Scholar 

  13. Doshi, N., J. N. Orje, B. Molins, J. W. Smith, S. Mitragotri, and Z. M. Ruggeri. Platelet mimetic particles for targeting thrombi in flowing blood. Adv. Mater. Weinh. 24:3864–3869, 2012.

    Article  Google Scholar 

  14. Eckstein, E. C., and F. Belgacem. Model of platelet transport in flowing blood with drift and diffusion terms. Biophys. J. 60:53–69, 1991.

    Article  Google Scholar 

  15. Flamm, M. H., and S. L. Diamond. Multiscale systems biology and physics of thrombosis under flow. Ann. Biomed. Eng. 40:2355–2364, 2012.

    Article  Google Scholar 

  16. Fogelson, A. L., and K. B. Neeves. Fluid mechanics of blood clot formation. Ann. Rev. Fluid Mech. 47:377–403, 2015.

    Article  Google Scholar 

  17. Hammon, J.W. Extracorporeal circulation. In: Cardiac Surgery in the Adult, 4th ed. New York: McGraw-Hill Medical, 2008, pp. 350–370.

  18. Han, X., R. Bibb, and R. Harris. Artificial vascular bifurcations—design and modelling. Proc. CIRP 49:14–18, 2016.

    Article  Google Scholar 

  19. Hansen, R. R., A. R. Wufsus, S. T. Barton, A. A. Onasoga, R. M. Johnson-Paben, and K. B. Neeves. High content evaluation of shear dependent platelet function in a microfluidic flow assay. Ann. Biomed. Eng. 41:250–262, 2013.

    Article  Google Scholar 

  20. Hathcock, J. J. Flow effects on coagulation and thrombosis. Arterioscler. Thromb. Vasc. Biol. 26:1729–1737, 2006.

    Article  Google Scholar 

  21. Holme, P. A., et al. Shear-induced platelet activation and platelet microparticle formation at blood flow conditions as in arteries with a severe stenosis. Arterioscler. Thromb. Vasc. Biol. 17:646–653, 1997.

    Article  Google Scholar 

  22. Hussain, M. A., S. Kar, and R. R. Puniyani. Relationship between power law coefficients and major blood constituents affecting the whole blood viscosity. J. Biosci. 24:329–337, 1999.

    Article  Google Scholar 

  23. Jackson, S. P., W. S. Nesbitt, and E. Westein. Dynamics of platelet thrombus formation. J. Thromb. Haemost. 7:17–20, 2009.

    Article  Google Scholar 

  24. Jain, A., A. Graveline, A. Waterhouse, A. Vernet, R. Flaumenhaft, and D. E. Ingber. A shear gradient-activated microfluidic device for automated monitoring of whole blood haemostasis and platelet function. Nat. Commun. 7:10176, 2016.

    Article  Google Scholar 

  25. Jain, A., and L. L. Munn. Biomimetic postcapillary expansions for enhancing rare blood cell separation on a microfluidic chip. Lab Chip 11:2941–2947, 2011.

    Article  Google Scholar 

  26. Johnson-Chavarria, E. M., M. Tanyeri, and C. M. Schroeder. A microfluidic-based hydrodynamic trap for single particles. J. Vis. Exp. 47:e2517, 2011.

    Google Scholar 

  27. Johnston, B. M., P. R. Johnston, S. Corney, and D. Kilpatrick. Non-Newtonian blood flow in human right coronary arteries: steady state simulations. J. Biomech. 37:709–720, 2004.

    Article  Google Scholar 

  28. Jønsson, V., J. E. Bock, and J. B. Nielsen. Significance of plasma skimming and plasma volume expansion. J. Appl. Physiol. 72:2047–2051, 1992.

    Google Scholar 

  29. Kang, E., D. H. Lee, C.-B. Kim, S. J. Yoo, and S.-H. Lee. A hemispherical microfluidic channel for the trapping and passive dissipation of microbubbles. J. Micromech. Microeng. 20:45009, 2010.

    Article  Google Scholar 

  30. Khandelwal, V., A. Dhiman, and L. Baranyi. Laminar flow of non-Newtonian shear-thinning fluids in a T-channel. Comput. Fluids 108:79–91, 2015.

    Article  MathSciNet  Google Scholar 

  31. Kniazeva, T., J. C. Hsiao, J. L. Charest, and J. T. Borenstein. A microfluidic respiratory assist device with high gas permeance for artificial lung applications. Biomed. Microdevices 13:315–323, 2011.

    Article  Google Scholar 

  32. Lasheras, J. C. The biomechanics of arterial aneurysms. Annu. Rev. Fluid Mech. 39:293–319, 2007.

    Article  MathSciNet  MATH  Google Scholar 

  33. Lee, S. Y. K., M. Wong, and Y. Zohar. Pressure losses in microchannels with bends. In: The 14th IEEE International Conference on Micro Electro Mechanical Systems, 2001. MEMS 2001, pp. 491–494, 2001.

  34. Lee, A. M., G. W. Tormoen, E. Kanso, O. J. T. McCarty, and P. K. Newton. Modeling and simulation of procoagulant circulating tumor cells in flow. Front. Oncol. 2:108, 2012.

    Google Scholar 

  35. Lei, M., C. Kleinstreuer, and G. A. Truskey. Numerical investigation and prediction of atherogenic sites in branching arteries. J. Biomech. Eng. 117:350–357, 1995.

    Article  Google Scholar 

  36. Liberale, C., et al. Integrated microfluidic device for single-cell trapping and spectroscopy. Sci. Rep. 3:1258, 2013.

    Google Scholar 

  37. Liu, L., et al. Inhibition of thrombin by antithrombin III and heparin cofactor II in vivo. Thromb. Haemost. 73:405–412, 1995.

    Google Scholar 

  38. Mackman, N. New insights into the mechanisms of venous thrombosis. J. Clin. Investig. 122:2331–2336, 2012.

    Article  Google Scholar 

  39. Maddala, J., W. S. Wang, S. A. Vanapalli, and R. Rengaswamy. Traffic of pairs of drops in microfluidic ladder networks with fore-aft structural asymmetry. Microfl. Nanofl. 14:337–344, 2013.

    Article  Google Scholar 

  40. Madershahian, N., R. Nagib, J. Wippermann, J. Strauch, and T. Wahlers. A simple technique of distal limb perfusion during prolonged femoro-femoral cannulation. J. Card. Surg. 21:168–169, 2006.

    Article  Google Scholar 

  41. McDonald, J. C., et al. Fabrication of microfluidic systems in poly(dimethylsiloxane). Electrophoresis 21:27–40, 2000.

    Article  Google Scholar 

  42. Meyer, A., M. Strüber, and S. Fischer. Advances in extracorporeal ventilation. Anesth. Clinics 26:381–391, 2008.

    Article  Google Scholar 

  43. Nesbitt, W. S., et al. A shear gradient–dependent platelet aggregation mechanism drives thrombus formation. Nat. Medicine 15:665–673, 2009.

    Article  Google Scholar 

  44. Nosovitsky, V. A., O. J. Ilegbusi, J. Jiang, P. H. Stone, and C. L. Feldman. Effects of curvature and stenosis-like narrowing on wall shear stress in a coronary artery model with phasic flow. Comp. Biomed. Res. 30:61–82, 1997.

    Article  Google Scholar 

  45. Pearson, T. C., and G. Wetherley-Mein. Vascular occlusive episodes and venous haematocrit in primary proliferative polycythaemia. Lancet 2:1219–1222, 1978.

    Article  Google Scholar 

  46. Perkkiö, J., L. J. Wurzinger, and H. Schmid-Schönbein. Plasma and platelet skimming at T-junctions. Thromb. Res. 45:517–526, 1987.

    Article  Google Scholar 

  47. Phillips, K. G., et al. Optical quantification of cellular mass, volume, and density of circulating tumor cells identified in an ovarian cancer patient. Front. Oncol. 2:72, 2012.

    Google Scholar 

  48. Popel, A. S., and P. C. Johnson. Microcirculation and hemorheology. Annu. Rev. Fluid Mech. 37:43–69, 2005.

    Article  MathSciNet  MATH  Google Scholar 

  49. Rana, K., B. J. Timmer, and K. B. Neeves. A combined microfluidic-microstencil method for patterning biomolecules and cells. Biomicrofluidics 8:56502, 2014.

    Article  Google Scholar 

  50. Replogle, R. L., H. J. Meiselman, and E. W. Merrill. Clinical implications of blood rheology studies. Circulation 36:148–160, 1967.

    Article  Google Scholar 

  51. Runyon, M. K., C. J. Kastrup, B. L. Johnson-Kerner, T. G. Van Ha, and R. F. Ismagilov. Effects of shear rate on propagation of blood clotting determined using microfluidics and numerical simulations. J. Am. Chem. Soc. 130:3458–3464, 2008.

    Article  Google Scholar 

  52. Shankaran, H., P. Alexandridis, and S. Neelamegham. Aspects of hydrodynamic shear regulating shear-induced platelet activation and self-association of von Willebrand factor in suspension. Blood 101:2637–2645, 2003.

    Article  Google Scholar 

  53. Tousi, N., B. Wang, K. Pant, M. F. Kiani, and B. Prabhakarpandian. Preferential adhesion of leukocytes near bifurcations is endothelium independent. Microvasc. Res. 80:384–388, 2010.

    Article  Google Scholar 

  54. Tsai, M., et al. In vitro modeling of the microvascular occlusion and thrombosis that occur in hematologic diseases using microfluidic technology. J. Clin. Investig. 122:408–418, 2012.

    Article  Google Scholar 

  55. Turitto, V. T., and H. R. Baumgartner. Platelet interaction with subendothelium in a perfusion system: physical role of red blood cells. Microvasc. Res. 9:335–344, 1975.

    Article  Google Scholar 

  56. Turitto, V. T., and H. J. Weiss. Red blood cells: their dual role in thrombus formation. Science 207:541–543, 1980.

    Article  Google Scholar 

  57. Vollmer, A. P., R. F. Probstein, R. Gilbert, and T. Thorsen. Development of an integrated microfluidic platform for dynamic oxygen sensing and delivery in a flowing medium. Lab Chip 5:1059–1066, 2005.

    Article  Google Scholar 

  58. Walker, C. P. R., and D. Royston. Thrombin generation and its inhibition: a review of the scientific basis and mechanism of action of anticoagulant therapies. Br. J. Anaesth. 88:848–863, 2002.

    Article  Google Scholar 

  59. Watts, T., M. Barigou, and G. B. Nash. Comparative rheology of the adhesion of platelets and leukocytes from flowing blood: why are platelets so small? Am. J. Physiol. Heart Circ. Physiol. 304:H1483–H1494, 2013.

    Article  Google Scholar 

  60. 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. USA 110:1357–1362, 2013.

    Article  Google Scholar 

  61. White-Adams, T. C., et al. Laminin promotes coagulation and thrombus formation in a factor XII-dependent manner. J. Thromb. Haemost. 8:1295–1301, 2010.

    Article  Google Scholar 

  62. Wu, W.-I., et al. Lung assist device: development of microfluidic oxygenators for preterm infants with respiratory failure. Lab Chip 13:2641–2650, 2013.

    Article  Google Scholar 

  63. You, J., L. Flores, M. Packirisamy, and I. Stiharu. Modeling the effect of channel bends on microfluidic flow. In: IASME Transactions Udine, pp. 144–151, 2005.

  64. Zheng, Y., J. Chen, and J. A. López. Flow-driven assembly of VWF fibres and webs in in vitro microvessels. Nat. Commun. 6:7858, 2015.

    Article  Google Scholar 

  65. Zilberman-Rudenko, J., et al. Biorheology of platelet activation in the bloodstream distal to thrombus formation. Cell. Mol. Bioeng. 9:1–13, 2016.

    Article  Google Scholar 

  66. Zilberman-Rudenko, J., et al. Coagulation factor XI promotes distal platelet activation and single platelet consumption in the bloodstream under shear flow. Arterioscler. Thromb. Vasc. Biol. 36:510–517, 2016.

    Article  Google Scholar 

  67. Zimmerman, W. B. J. Microfluidics: History, Theory and Applications. New York: Springer, 2006.

    Google Scholar 

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Acknowledgments

We thank Dr. András Gruber for insightful comments and Chantal Wiesenekker for technical assistance. This work was supported by West Virginia University startup funds awarded to J. Maddala and by grants from the National Institutes of Health (R01HL101972, R01GM116184, R44HL126235). O.J.T. McCarty is an American Heart Association Established Investigator (13EIA12630000).

Conflict of interest

J. Zilberman-Rudenko, J.L. Sylman, H.H.S. Lakshman, O.J.T. McCarty and J. Maddala declare no competing financial interests.

Human and Animal Rights and Informed Consent

All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2000 (5). Informed consent was obtained from all patients for being included in the study. All human subject research was carried out in accordance with institutional guidelines approved by the Oregon Health & Science University Institutional Review Board. No animal studies were carried out by the authors for this article.

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Authors and Affiliations

  1. Biomedical Engineering, School of Medicine, Oregon Health and Science University, Portland, OR, USA

    Jevgenia Zilberman-Rudenko, Joanna L. Sylman, Owen J. T. McCarty & Jeevan Maddala

  2. Chemical and Biomedical Engineering, West Virginia University, Morgantown, WV, USA

    Hari H. S. Lakshmanan & Jeevan Maddala

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  1. Jevgenia Zilberman-Rudenko
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Correspondence to Jeevan Maddala.

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Associate Editor Michael R. King oversaw the review of this article.

O. J. T. McCarty and J. Maddala are co-senior authors.

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Supplemental Video 1

Initial blood flow dynamics within the ladder microfluidic device. Recalcified whole human blood was perfused at a 2 µL/min flow rate through a PDMS-ladder network coated with collagen and tissue factor; real-time video of the blood flow was recorded differential using interference contrast, DIC, microscopy. Supplementary material 1 (WMV 4603 kb)

Supplemental Video 2

Blood flow dynamics within the ladder microfluidic device after 10 min of blood perfusion. Recalcified whole human blood was perfused at a 2 µL/min flow rate through a PDMS-ladder network coated with collagen and tissue factor; real-time video of the blood flow was recorded differential using interference contrast, DIC, microscopy at the 10 min time point. Supplementary material 2 (WMV 447 kb)

Supplemental Fig. 1

Quantification of the spatial distribution of thrombus formation. DiOC6-labeled whole human blood was perfused at a 2 µL/min flow rate through a PDMS-ladder network coated with collagen and tissue factor; images of thrombus formation were recorded using differential interference contrast, DIC microscopy. The thrombus surface area was quantified for 5 experiments using ImageJ. Supplementary material 3 (TIFF 976 kb)

Supplementary material 4 (PDF 110 kb)

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Zilberman-Rudenko, J., Sylman, J.L., Lakshmanan, H.H.S. et al. Dynamics of Blood Flow and Thrombus Formation in a Multi-Bypass Microfluidic Ladder Network. Cel. Mol. Bioeng. 10, 16–29 (2017). https://doi.org/10.1007/s12195-016-0470-7

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