Baseline Platelet Activation and Reactivity in Patients with Critical Limb Ischemia

Peter Paul Wisman, Martin Teraa, Gert Jan de Borst, Marianne C Verhaar, Mark Roest, Frans L Moll, Peter Paul Wisman, Martin Teraa, Gert Jan de Borst, Marianne C Verhaar, Mark Roest, Frans L Moll

Abstract

Background: Patients with critical limb ischemia (CLI) have a high risk to develop cardiovascular events (CVE). We hypothesized that in CLI patients platelets would display increased baseline activation and reactivity.

Objectives: We investigated baseline platelet activation and platelet reactivity in patients with CLI.

Patients/methods: In this study baseline platelet activation and platelet reactivity in response to stimulation of all major platelet activation pathways were determined in 20 CLI patients (11 using aspirin and 9 using vitamin K-antagonists) included in the Juventas-trial (clinicaltrials.gov NCT00371371) and in 17 healthy controls. Platelet activation was quantified with flow cytometric measurement of platelet P-selectin expression and fibrinogen binding.

Results: CLI patients not using aspirin showed higher baseline platelet activation compared to healthy controls. Maximal reactivity to stimulation of the collagen and thrombin activation pathway was decreased in CLI patients compared to healthy controls. In line, attenuated platelet reactivity to stimulation of multiple activation pathways was associated with several traditional risk factors for cardiovascular disease.

Conclusions: Baseline platelet activation was increased in CLI patients, whereas the reactivity of circulating platelets to several stimulatory agents is decreased. Reactivity of platelets was inversely correlated with cardiovascular risk factors.

Conflict of interest statement

Competing Interests: The authors certify that they have no affiliation with or financial involvement in any organization or entity with a direct financial interest in the subject matter or materials discussed in the manuscript (e.g., employment, consultancies, stock ownership, honoraria).

Figures

Fig 1. Baseline platelet activation of CLI…
Fig 1. Baseline platelet activation of CLI A- patients versus healthy controls and CLI A- patients versus CLI A+ patients.
MFI for bound fibrinogen or P-selectin expression without stimulation, stratified for CLI A- patients versus healthy controls and CLI A- patients versus CLI A+ patients. MFI median fluorescence intensity, * p<0.05.
Fig 2. Platelet reactivity of CLI A-…
Fig 2. Platelet reactivity of CLI A- patients versus healthy controls.
MFI for fibrinogen binding or P-selectin expression per agonist, stratified for CLI A- patients versus healthy controls. CLI A- CLI patients not treated with aspirin, MFI median fluorescence intensity, CVX Convulxin, ADP Adenosine Diphosphate, TRAP Thrombin receptor agonist SFLLRN, Tx Thromboxane receptor agonist, Curve dose-response curves of each group differ significantly (p<0.05), Max maximal platelet reactivity of each group differ significantly (p<0.05), EC50 half maximal effective concentration of each group differ significantly (p<0.05).
Fig 3. Platelet reactivity of CLI A-…
Fig 3. Platelet reactivity of CLI A- patients versus CLI A+ patients.
MFI for fibrinogen binding or P-selectin expression per agonist, stratified for CLI A- patients versus CLI A+ patients. CLI A- CLI patients not treated with aspirin, CLI A+ CLI patients treated with aspirin, MFI median fluorescence intensity, CVX Convulxin, ADP Adenosine Diphosphate, TRAP Thrombin receptor agonist SFLLRN, Tx Thromboxane receptor agonist, Curve curves for each group differ significantly (p<0.05), Max maximal platelet reactivity for each group differ significantly (p<0.05).

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