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A shear gradient–dependent platelet aggregation mechanism drives thrombus formation

Nature Medicine volume 15, pages 665673 (2009) | Download Citation

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Abstract

Platelet aggregation at sites of vascular injury is essential for hemostasis and arterial thrombosis. It has long been assumed that platelet aggregation and thrombus growth are initiated by soluble agonists generated at sites of vascular injury. By using high-resolution intravital imaging techniques and hydrodynamic analyses, we show that platelet aggregation is primarily driven by changes in blood flow parameters (rheology), with soluble agonists having a secondary role, stabilizing formed aggregates. We find that in response to vascular injury, thrombi initially develop through the progressive stabilization of discoid platelet aggregates. Analysis of blood flow dynamics revealed that discoid platelets preferentially adhere in low-shear zones at the downstream face of forming thrombi, with stabilization of aggregates dependent on the dynamic restructuring of membrane tethers. These findings provide insight into the prothrombotic effects of disturbed blood flow parameters and suggest a fundamental reinterpretation of the mechanisms driving platelet aggregation and thrombus growth.

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Acknowledgements

We thank Z. Ruggeri, H.H. Salem, R. Andrews and B. Kile for helpful feedback on the work; H. Blackburn and G. Rosengarten for advice and input on the CFD analysis; C. Nguyen for technical assistance in the analysis of in vivo blood flow rates; SciTech Proprietary Ltd. and Andor Proprietary Ltd. for the generous loan of a DV897CS EMCCD camera; M. Hickey (Monash University Department of Medicine, Monash, Australia) and A. Issekutz (Dalhousie University, Halifax, Canada) for providing the P-selectin–specific antibody; and C. Gachet (INSERM, Strasbourg, France) for P2Y1−/− mice. This work was supported by project funding from the National Health and Medical Research Council of Australia and the Australian Research Council. E.W. was supported by the National Heart Foundation of Australia.

Author information

Author notes

    • Warwick S Nesbitt
    •  & Erik Westein

    These authors contributed equally to this work.

Affiliations

  1. The Australian Centre for Blood Diseases, Monash University, Alfred Medical Research and Educational Precinct, Melbourne, Victoria, Australia.

    • Warwick S Nesbitt
    • , Erik Westein
    • , Jia Fu
    •  & Shaun P Jackson
  2. Microelectronics and Materials Technology Centre, School of Electrical and Computer Engineering, RMIT University, Melbourne, Victoria, Australia.

    • Francisco Javier Tovar-Lopez
    •  & Arnan Mitchell
  3. Department of Mechanical Engineering, Monash University, Clayton, Victoria, Australia.

    • Elham Tolouei
    •  & Josie Carberry
  4. Division of Biological Engineering, Monash University, Clayton, Victoria, Australia.

    • Andreas Fouras

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Contributions

W.S.N. designed the study and experiments, performed the in vitro platelet and micro-PIV experiments, codesigned vascular mimetics, performed microchannel perfusion experiments, analyzed data, supervised the study and cowrote the manuscript. E.W. designed experiments, performed the in vivo and in vitro experiments, analyzed data and assisted with manuscript preparation. F.J.T.-L. codesigned and fabricated vascular mimetics (microchannels), performed microchannel perfusion experiments and performed CFD simulations. E.T. performed and analyzed the micro-PIV experiments. J.F. performed in vivo experiments. A.M. codesigned the vascular mimetics and supervised their fabrication. J.C. supervised the micro-PIV analysis. A.F. designed and supervised the micro-PIV analysis and formulated and coded the micro-PIV analysis software. S.P.J. supervised the study and cowrote the manuscript.

Corresponding authors

Correspondence to Warwick S Nesbitt or Shaun P Jackson.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figs. 1–6 and Supplementary Methods

Videos

  1. 1.

    Supplementary Video 1

    Rheology-driven platelet aggregation in vivo.

  2. 2.

    Supplementary Video 2

    Shear microgradient-induced platelet aggregation in vitro.

  3. 3.

    Supplementary Video 3

    Discoid aggregate formation independent of ADP, TXA2 and thrombin.

  4. 4.

    Supplementary Video 4

    Discoid platelet aggregation and aggregate consolidation in vivo.

  5. 5.

    Supplementary Video 5

    Platelet tethering interactions in vivo and in vitro.

  6. 6.

    Supplementary Video 6

    Intracellular calcium flux during platelet tether restructuring.

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DOI

https://doi.org/10.1038/nm.1955

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