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Thromb Haemost 2016; 115(02): 333-343
DOI: 10.1160/th15-04-0300
Cellular Haemostasis and Platelets
Schattauer GmbH

Matrix metalloproteinase-2 enhances platelet deposition on collagen under flow conditions

Giuseppe Guglielmini   *
1   Department of Medicine, Section of Internal and Cardiovascular Medicine, University Perugia, Italy
,
Viviana Appolloni   *
1   Department of Medicine, Section of Internal and Cardiovascular Medicine, University Perugia, Italy
,
Stefania Momi
1   Department of Medicine, Section of Internal and Cardiovascular Medicine, University Perugia, Italy
,
Philip G. de Groot
2   Department of Clinical Chemistry and Hematology, University Medical Centre, Utrecht, The Netherlands
,
Monica Battiston
3   Department of Translational Research, Stem Cell Unit, National Cancer Institute CRO-IRCCS, Aviano, Italy
,
Luigi de Marco
3   Department of Translational Research, Stem Cell Unit, National Cancer Institute CRO-IRCCS, Aviano, Italy
4   Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla ,California, USA
,
Emanuela Falcinelli
1   Department of Medicine, Section of Internal and Cardiovascular Medicine, University Perugia, Italy
,
Paolo Gresele
1   Department of Medicine, Section of Internal and Cardiovascular Medicine, University Perugia, Italy
› Author Affiliations Financial support: This study was supported in part by grants from Fondazione Cassa di Risparmio di Perugia (Protocol # 2014.0083.021), from MIUR (Protocol # 2012773NE3) to P. G., and from Fondazione Umbero Veronesi to E. F., and in part by an Erasmus Placement grant to V. A. and by a grant to L. D. M. from the Italian Ministry of Health RF-2010–2316198.
Further Information

Publication History

Received: 10 April 2015

Accepted after major revision: 18 September 2015

Publication Date:
22 November 2017 (online)

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Summary

Platelets contain and release matrix metalloproteinase-2 (MMP-2) that in turn potentiates platelet aggregation. Platelet deposition on a damaged vascular wall is the first, crucial, step leading to thrombosis. Little is known about the effects of MMP-2 on platelet activation and adhesion under flow conditions. We studied the effect of MMP-2 on shear-dependent platelet activation using the O’Brien filtration system, and on platelet deposition using a parallel-plate perfusion chamber. Preincubation of human whole blood with active MMP-2 (50 ng/ml, i. e. 0.78 nM) shortened filter closure time (from 51.8 ± 3.6 sec to 40 ± 2.7 sec, p< 0.05) and increased retained platelets (from 72.3 ± 2.3 % to 81.1 ± 1.8 %, p< 0.05) in the O’Brien system, an effect prevented by a specific MMP-2 inhibitor. High shear stress induced the release of MMP-2 from platelets, while TIMP-2 levels were not significantly reduced, therefore, the MMP-2/TIMP-2 ratio increased significantly showing enhanced MMP-2 activity. Preincubation of whole blood with active MMP-2 (0.5 to 50 ng/ml, i.e 0.0078 to 0.78 nM) increased dose-dependently human platelet deposition on collagen under high shear-rate flow conditions (3000 sec-1) (maximum +47.0 ± 11.9 %, p< 0.05, with 50 ng/ml), while pre-incubation with a MMP-2 inhibitor reduced platelet deposition. In real-time microscopy studies, increased deposition of platelets on collagen induced by MMP-2 started 85 sec from the beginning of perfusion, and was abolished by a GPIIb/IIIa antagonist, while MMP-2 had no effect on platelet deposition on fibrinogen or VWF. Confocal microscopy showed that MMP-2 enhances thrombus volume (+20.0 ± 3.0 % vs control) rather than adhesion. In conclusion, we show that MMP-2 potentiates shear-induced platelet activation by enhancing thrombus formation.

Supplementary Material to this article is available online at www.thrombosis-online.com.

Keywords

Matrix metalloproteinases - platelet adhesion - shear stress - TIMP-2

* These authors contributed equally to this study.


 

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