Use of Thrombodynamics for revealing the participation of platelet, erythrocyte, endothelial, and monocyte microparticles in coagulation activation and propagation.

E N Lipets,E N Lipets,K V Losenkova,O A Antonova,A V Mazurov,F I Ataullakhanov,F I Ataullakhanov,F I Ataullakhanov,F I Ataullakhanov,O N Shustova,Arijit Biswas

doi:10.1371/journal.pone.0227932

E N Lipets, E N Lipets + Show 9 more

Open Access

https://doi.org/10.1371/journal.pone.0227932

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Abstract

Background and objectiveFor many pathological states, microparticles are supposed to be one of the causes of hypercoagulation. Although there are some indirect data about microparticles participation in coagulation activation and propagation, the integral hemostasis test Thrombodynamics allows to measure micropaticles participation in these two coagulation phases directly. Demonstrates microparticles participation in coagulation activation by influence on the appearance of coagulation centres in the plasma volume and the rate of clot growth from the surface with immobilized tissue factor.Methods: Microparticles were obtained from platelets and erythrocytes by stimulation with thrombin receptor-activating peptide (SFLLRN) and calcium ionophore (A23187), respectively, from monocytes, endothelial HUVEC culture and monocytic THP cell culture by stimulation with lipopolysaccharides. Microparticles were counted by flow cytometry and titrated in microparticle-depleted normal plasma in the Thrombodynamics test.ResultsMonocyte microparticles induced the appearance of clotting centres through the TF pathway at concentrations approximately 100-fold lower than platelet and erythrocyte microparticles, which activated plasma by the contact pathway. For endothelial microparticles, both activation pathways were essential, and their activity was intermediate. Monocyte microparticles induced plasma clotting by the appearance of hundreds of clots with an extremely slow growth rate, while erythrocyte microparticles induced the appearance of a few clots with a growth rate similar to that from surface covered with high-density tissue factor. Patterns of clotting induced by platelet and endothelial microparticles were intermediate. Platelet, erythrocyte and endothelial microparticles impacts on the rate of clot growth from the surface with tissue factor did not differ significantly within the 0–200·103/ul range of microparticles concentrations. However, at concentrations greater than 500·103/ul, erythrocyte microparticles increased the stationary clot growth rate to significantly higher levels than do platelet microparticles or artificial phospholipid vesicles consisting of phosphatidylcholine and phosphatidylserine.ConclusionMicroparticles of different origins demonstrated qualitatively different characteristics related to coagulation activation and propagation.

Highlights

Cell destruction or activation leads to microparticles (MPs) shedding
Monocyte microparticles induced the appearance of clotting centres through the tissue factor (TF) pathway at concentrations approximately 100-fold lower than platelet and erythrocyte microparticles, which activated plasma by the contact pathway
Monocyte microparticles induced plasma clotting by the appearance of hundreds of clots with an extremely slow growth rate, while erythrocyte microparticles induced the appearance of a few clots with a growth rate similar to that from surface covered with high-density tissue factor

Summary

Introduction

Cell destruction or activation leads to microparticles (MPs) shedding. In the blood of normal donors, more than 80% of MPs are derived from platelets [1,2]. There are a number of clinical works where the MPs concentration is shown to increase in pathological states associated with elevated thrombotic risk. Increased tissue factor-bearing MPs concentration [13,14] or activity [15,16] was associated with venous thromboembolic events in patients with cancer. A number of prospective studies have appeared that examine the role of increased concentrations of MPs in cancer and recurrent thrombosis, but their results are contradictory [5,17]. Prospective studies are few: EMPs concentration increase was an independent predictor of cardiovascular complications in patients with heart failure, type II diabetes, and end-stage renal failure [18,19,20]. Microparticles were counted by flow cytometry and titrated in microparticle-depleted normal plasma in the Thrombodynamics test

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