The inhibitory effect of antiplatelet drugs on spontaneous platelet aggregation on glass surfaces: an analysis based on microscopic three-dimensional topography

In vitro assessment of the inhibitory effect of antiplatelet drugs on platelet aggregation is frequently employed to guide personalized antiplatelet therapy in clinical practice. However, existing methods for detecting platelet aggregation rely heavily on high concentrations of exogenous agonists, which may obscure part of the inhibitory effect of antiplatelet drugs and lead to an underestimation of their effects. This study validates a novel analytical strategy for evaluating the effects of antiplatelet drugs by quantifying the microscopic three-dimensional morphological parameters of platelet aggregates formed through spontaneous aggregation on a glass surface. Heparin-anticoagulated platelet-rich plasma (PRP) was applied to a glass surface to induce spontaneous platelet aggregation. The microscopic three-dimensional morphology of platelet aggregates was characterized using a laser three-dimensional microscopic imaging system, and platelet aggregation function was assessed based on the volume parameter (Vol) and cross-sectional area parameter (CS-area) of the aggregates. The results demonstrated that platelets could spontaneously aggregate on the glass surface under the participation of plasma proteins and CA²⁺. Aspirin (80 μM) significantly reduced Vol but had no significant effect on CS-area. Ticagrelor, eptifibatide, and tirofiban dose-dependently decreased both Vol and CS-area. High concentrations of eptifibatide (4 μM) and tirofiban (4 μM) completely inhibited platelet adhesion and aggregation. The combination of aspirin (20 μM) and ticagrelor (0.5 μM) synergistically suppressed platelet aggregation behavior. GPIb-IX-von Willebrand factor (vWF) inhibitors (4 μM) and indomethacin (4 μM) significantly reduced both Vol and CS-area, with a smaller reduction in CS-area compared to Vol. In patients, aspirin alone significantly reduced Vol, while clopidogrel, aspirin combined with clopidogrel, and Xuesaitong significantly decreased both Vol and CS-area. Our novel analytical strategy is capable of distinguishing the pharmacological effects of various antiplatelet agents without the need for exogenous agonists, suggesting that this system may aid in the determination of the appropriate type and dose of the antiplatelet agent in the clinical setting.

Antiplatelet drugs; Glass; Microscopic three-dimensional morphology; Platelet aggregation.