Platelets Drive Thrombus Propagation in a Hematocrit and Glycoprotein VI-Dependent Manner in an In Vitro Venous Thrombosis Model

Arterioscler Thromb Vasc Biol. 2018 May;38(5):1052-1062. doi: 10.1161/ATVBAHA.118.310731. Epub 2018 Feb 22.


Objective: The objective of this study was to measure the role of platelets and red blood cells on thrombus propagation in an in vitro model of venous valvular stasis.

Approach and results: A microfluidic model with dimensional similarity to human venous valves consists of a sinus distal to a sudden expansion, where for sufficiently high Reynolds numbers, 2 countercurrent vortices arise because of flow separation. The primary vortex is defined by the points of flow separation and reattachment. A secondary vortex forms in the deepest recess of the valve pocket characterized by low shear rates. An initial fibrin gel formed within the secondary vortex of a tissue factor-coated valve sinus. Platelets accumulated at the interface of the fibrin gel and the primary vortex. Red blood cells at physiological hematocrits were necessary to provide an adequate flux of platelets to support thrombus growth out of the valve sinus. A subpopulation of platelets that adhered to fibrin expose phosphatidylserine. Platelet-dependent thrombus growth was attenuated by inhibition of glycoprotein VI with a blocking Fab fragment or D-dimer.

Conclusions: A 3-step process regulated by hemodynamics was necessary for robust thrombus propagation: First, immobilized tissue factor initiates coagulation and fibrin deposition within a low flow niche defined by a secondary vortex in the pocket of a model venous valve. Second, a primary vortex delivers platelets to the fibrin interface in a red blood cell-dependent manner. Third, platelets adhere to fibrin, activate through glycoprotein VI, express phosphatidylserine, and subsequently promote thrombus growth beyond the valve sinus and into the bulk flow.

Keywords: blood platelets; erythrocytes; hematocrit; hemorheology; phosphatidylserines.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Video-Audio Media

MeSH terms

  • Blood Coagulation*
  • Blood Flow Velocity
  • Blood Platelets / metabolism*
  • Erythrocytes / metabolism*
  • Fibrin / metabolism
  • Hematocrit
  • Hemodynamics
  • Humans
  • Lab-On-A-Chip Devices
  • Microfluidic Analytical Techniques / instrumentation
  • Phosphatidylserines / blood
  • Platelet Membrane Glycoproteins / metabolism*
  • Signal Transduction
  • Stress, Mechanical
  • Thromboplastin / metabolism
  • Venous Thrombosis / blood*
  • Venous Thrombosis / pathology
  • Venous Thrombosis / physiopathology
  • Venous Valves / metabolism*
  • Venous Valves / pathology
  • Venous Valves / physiopathology


  • Phosphatidylserines
  • Platelet Membrane Glycoproteins
  • platelet membrane glycoprotein VI
  • Fibrin
  • Thromboplastin