Platelet Adhesion and Thrombus Formation in Microchannels: The Effect of Assay-Dependent Variables

Int J Mol Sci. 2020 Jan 23;21(3):750. doi: 10.3390/ijms21030750.

Abstract

Microfluidic flow chambers (MFCs) allow the study of platelet adhesion and thrombus formation under flow, which may be influenced by several variables. We developed a new MFC, with which we tested the effects of different variables on the results of platelet deposition and thrombus formation on a collagen-coated surface.

Methods: Whole blood was perfused in the MFC over collagen Type I for 4 min at different wall shear rates (WSR) and different concentrations of collagen-coating solutions, keeping blood samples at room temperature or 37 °C before starting the experiments. In addition, we tested the effects of the antiplatelet agent acetylsalicylic acid (ASA) (antagonist of cyclooxygenase-1, 100 µM) and cangrelor (antagonist of P2Y12, 1 µM).

Results: Platelet deposition on collagen (I) was not affected by the storage temperature of the blood before perfusion (room temperature vs. 37 °C); (II) was dependent on a shear rate in the range between 300/s and 1700/s; and (III) was influenced by the collagen concentration used to coat the microchannels up to a value of 10 µg/mL. ASA and cangrelor did not cause statistically significant inhibition of platelet accumulation, except for ASA at low collagen concentrations.

Conclusions: Platelet deposition on collagen-coated surfaces is a shear-dependent process, not influenced by the collagen concentration beyond a value of 10 µg/mL. However, the inhibitory effect of antiplatelet drugs is better observed using low concentrations of collagen.

Keywords: antiplatelet agents; flow assays; microfluidics; platelet adhesion; platelet aggregation; platelets; shear rate; thrombus formation.

MeSH terms

  • Adenosine Monophosphate / analogs & derivatives*
  • Adenosine Monophosphate / pharmacology
  • Aspirin / pharmacology*
  • Blood Platelets / drug effects
  • Collagen
  • Cyclooxygenase Inhibitors / pharmacology
  • Humans
  • Male
  • Microfluidics / instrumentation*
  • Platelet Adhesiveness / drug effects*
  • Platelet Aggregation / drug effects*
  • Platelet Aggregation Inhibitors / pharmacology*
  • Purinergic P2Y Receptor Antagonists / pharmacology
  • Thrombosis / diagnostic imaging
  • Thrombosis / etiology*
  • Thrombosis / metabolism
  • Young Adult

Substances

  • Cyclooxygenase Inhibitors
  • Platelet Aggregation Inhibitors
  • Purinergic P2Y Receptor Antagonists
  • Adenosine Monophosphate
  • cangrelor
  • Collagen
  • Aspirin