Coagulation on Endothelial Cells: The Underexposed Part of Virchow’s Triad

Irma L.A. Geenen; Mark J. Post; Daniel G.M. Molin; Geert W.H. Schurink; Jos G. Maessen; Rene v. Oerle; Hugo ten Cate; Henri M.H. Spronk

doi:10.1160/TH12-04-0269

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2012; 108(05): 863-871
DOI: 10.1160/TH12-04-0269

Theme Issue Article

Schattauer GmbH

Coagulation on Endothelial Cells: The Underexposed Part of Virchow’s Triad

Authors

Irma L.A. Geenen

¹Department of Physiology, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University Medical Center, Maastricht, The Netherlands

²Department of General Surgery, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University Medical Center, Maastricht, The Netherlands
Mark J. Post

¹Department of Physiology, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University Medical Center, Maastricht, The Netherlands
Daniel G.M. Molin

¹Department of Physiology, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University Medical Center, Maastricht, The Netherlands
Geert W.H. Schurink

²Department of General Surgery, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University Medical Center, Maastricht, The Netherlands
Jos G. Maessen

³Department of Cardiothoracic Surgery, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University Medical Center, Maastricht, The Netherlands
Rene v. Oerle

⁴Department of Internal Medicine, Laboratory of Clinical Thrombosis and Haemostasis, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University Medical Center, Maastricht, The Netherlands
Hugo ten Cate

⁴Department of Internal Medicine, Laboratory of Clinical Thrombosis and Haemostasis, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University Medical Center, Maastricht, The Netherlands
Henri M.H. Spronk

⁴Department of Internal Medicine, Laboratory of Clinical Thrombosis and Haemostasis, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University Medical Center, Maastricht, The Netherlands

Abstract

Summary

The process of thrombin generation involves numerous plasma proteases and cofactors. Interaction with the vessel wall, in particular endothelial cells (ECs), influences this process but data on this interaction is limited. We evaluated thrombin generation on EA.hy926, human coronary arterial ECs (HCAECs) and patient-derived human venous ECs (HVECs) by means of a modified calibrated automated thrombogram (CAT) method and especially looked into contribution of the intrinsic and extrinsic pathways. Thrombin generation was measured in presence of confluent ECs with normal pooled and factor XII-deficient (FXII-deficient) platelet-poor plasma, with/without active site inhibited factor VIIa (ASIS) to block the extrinsic pathway and corn trypsin inhibitor for blocking contact activation (intrinsic pathway). Fetal bovine serum (FBS) was removed from culture conditions as FXIIa from the serum retained on ECs apparently, thereby inducing strong contact activation. In serum-free conditions, EA.hy926 and patient-derived HVECs induced thrombin generation mainly via the contact activation pathway with minor influence of ASIS on peak height and very low thrombin generation curves in FXII-deficient plasma. HVECs derived from coronary arterial bypass graft (CABG) patients showed increased thrombin generation compared to control patients, which could be ascribed to increased contact activation. Contribution of the extrinsic pathway on patient-derived ECs was limited. We conclude that the CAT method in combination with serum-free cultured ECs offers a valuable high-throughput method to evaluate endothelial influences on thrombin generation, which appears to involve predominantly contact activation on ECs. Contact activation-mediated thrombin generation was increased on ECs from CABG patients compared to controls.

Keywords

Endothelial cells - thrombin generation - cell culture - contact activation - extrinsic pathway

Full Text

References

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