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Endothelial Cell-derived Extracellular Vesicles Size-dependently Exert Procoagulant Activity Detected by
Wolfgang Holnthoner1,2, Cornelia Bonstingl3,4, Carina Hromada3,4
1Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research Centre, Vienna, Austria. wolfgang.holnthoner@trauma.lbg.ac.at.
Scientific Reports
|June 18, 2017
Summary
Endothelial cells release tissue factor-bearing extracellular vesicles (EVs) that promote blood clotting. These procoagulant EVs, varying in size, are released upon TNF-α stimulation, impacting hemostasis.
Area of Science:
- Vascular Biology
- Hemostasis and Thrombosis
- Extracellular Vesicles
Background:
- Endothelial cells (ECs) regulate hemostasis by releasing procoagulant and anticoagulant factors.
- Previous research indicated cultured ECs release procoagulant mediators, an effect neutralized by anti-tissue factor (TF) antibodies.
- The specific role of endothelial-derived extracellular vesicles (EVs) in releasing TF and their procoagulant activity remained unclear.
Purpose of the Study:
- To investigate if tissue factor (TF) is released by endothelial-derived extracellular vesicles (EVs).
- To determine which size fraction of released EVs exhibits the most significant procoagulant properties.
- To elucidate the role of TF-bearing EVs in endothelial cell-mediated hemostasis.
Main Methods:
- Endothelial cells (ECs) were stimulated with tumor-necrosis factor-α (TNF-α).
- Supernatants underwent differential centrifugation to isolate larger and smaller EVs.
- EVs were characterized using nanoparticle tracking analysis (NTA) and flow cytometry.
- Procoagulant activity was assessed by mixing EVs with human blood and analyzing via thromboelastometry.
Main Results:
- Stimulated ECs released EVs that exerted a significant procoagulant stimulus in blood.
- This procoagulant effect was partially reversed by an anti-TF antibody, confirming TF's role.
- TF activity was detected in the isolated EV fractions.
- Both smaller and larger EV fractions demonstrated procoagulant potential.
Conclusions:
- Endothelial cells release TF-bearing EVs of varying sizes upon TNF-α stimulation.
- These endothelial-derived EVs possess procoagulant properties, contributing to hemostasis.
- The findings highlight TF-bearing EVs as key mediators in EC-driven coagulation.
