Abstract
Platelet aggregometry assays are generally used for the analysis of platelet function but can also be adapted for further research and therapy focused applications. This method describes the procedures for the preparation of human platelet-rich plasma (PRP) and platelet-poor plasma (PPP) for the assessment of human platelet aggregation induced by agonists such as platelet-activating factor (PAF), thrombin, collagen, adenosine diphosphate (ADP), arachidonic acid, etc.•This method can be applied in vitro to evaluate the aggregatory effects of these agonists and to assess the antiaggregatory effects of several bioactive antiplatelet agents (compounds of natural or pharmacological origin) in human PRP.•This versatile method can be used in both basic and clinical research for the assessment of platelet aggregation (a major cardiovascular risk factor), platelet agonists, and inhibitors, in physiological or pathological conditions.•This method can be adapted to assess platelet activity in postprandial and intervention studies ex vivo.
Highlights
Human platelets are crucially involved in both normal haemostasis, pathological bleeding, and thrombosis
4 A linear relationship exists between the concentrations of an agonist (i.e. platelet-activating factor (PAF) or thrombin) within a specific range that induces platelet aggregation within the 20%–80% of the maximum-reversible platelet aggregation of hPRP
2 The aggregation curve of the maximum-reversible platelet aggregation of platelet-rich plasma (PRP) (100% aggregation of platelets) induced by a specific concentration of an agonist (PAF or thrombin) in the absence of any antiplatelet agent is determined as the 0% inhibition
Summary
Human platelets are crucially involved in both normal haemostasis, pathological bleeding, and thrombosis. Notes on assessing the inhibitory effects of potential antiaggregatory compounds: The application of LTA for evaluating the inhibitory effects (IC50 values) of an antiplatelet agent against platelet aggregation of PRP induced by PAF or thrombin is important, since it provides information concerning the pathway that this antiplatelet compound can affect or inhibit (either that of the PAF/PAF-receptor pathway or that of the Thrombin/PAR-1 pathway, or both), and at which concentration these effects are induced [1,12,13,14,16,17,18,19] These in vitro results are crucial for determining further research, either in cells and cell-related responses in vitro or ex vivo/in vivo in humans/animal-models. For instance if testing with PAF and/or PAF-like molecules with agonistic effect [2], pharmacological inhibitors such as Rupatadine or CV-3988 may be used, or inhibitors of natural origin such as Ginkgolide B (i.e. BN52021) to assess and compare to the antiplatelet agent being tested as previously described [23,24]
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