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.
Highlights
Platelets play a central pathogenic role in thrombosis; they aggregate at sites of atherosclerotic plaques, forming thrombi that can occlude the lumen of the artery [1,2,3]
The current study aims at investigating a number of critical aspects in the study of platelet adhesion and thrombus formation with Microfluidic flow chambers (MFCs): The effect of the storage temperature of the blood samples before testing; the influences of wall shear rate and of the concentration of the adhesive protein collagen in the surface coating solution; and the inhibitory effect of some antiplatelet drugs
We measured the effect of blood storage temperature at room temperature (RT) and 37 ◦C on platelet accumulation on collagen (200 μg/mL)-coated channels, at 300/s, 1100/s and 1700/s wall shear rates (Figure 1)
Summary
Platelets play a central pathogenic role in thrombosis; they aggregate at sites of atherosclerotic plaques, forming thrombi that can occlude the lumen of the artery [1,2,3]. Despite these studies and some attempts to define common protocols [18,23], no real standardization has been achieved and the effect of a number of assay-dependent variables still remains to be fully elucidated Given this background, the current study aims at investigating a number of critical aspects in the study of platelet adhesion and thrombus formation with MFCs: The effect of the storage temperature of the blood samples before testing; the influences of wall shear rate and of the concentration of the adhesive protein collagen in the surface coating solution; and the inhibitory effect of some antiplatelet drugs
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