Endothelial Cell-derived Extracellular Vesicles Size-dependently Exert Procoagulant Activity Detected by Thromboelastometry
Abstract Endothelial cells (ECs) are major modulators of hemostasis by expressing and releasing pro- and anticoagulant mediators into the circulation. Previous studies showed that cultured ECs release procoagulant mediators into cell culture supernatants as evidenced by the reduction of viscoelastic...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
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Nature Portfolio
2017-06-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-017-03159-0 |
| _version_ | 1830288499429867520 |
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| author | Wolfgang Holnthoner Cornelia Bonstingl Carina Hromada Severin Muehleder Johannes Zipperle Stefan Stojkovic Heinz Redl Johann Wojta Herbert Schöchl Johannes Grillari Sylvia Weilner Christoph J. Schlimp |
| author_facet | Wolfgang Holnthoner Cornelia Bonstingl Carina Hromada Severin Muehleder Johannes Zipperle Stefan Stojkovic Heinz Redl Johann Wojta Herbert Schöchl Johannes Grillari Sylvia Weilner Christoph J. Schlimp |
| author_sort | Wolfgang Holnthoner |
| collection | DOAJ |
| description | Abstract Endothelial cells (ECs) are major modulators of hemostasis by expressing and releasing pro- and anticoagulant mediators into the circulation. Previous studies showed that cultured ECs release procoagulant mediators into cell culture supernatants as evidenced by the reduction of viscoelastic clotting time. This effect was reversed with an anti-tissue factor antibody. Here, we aimed to investigate whether tissue factor (TF) was released by endothelial-derived extracellular vesicles (EVs) and which portion of the released vesicles displays the most prominent procoagulant properties. After stimulation of ECs with tumor-necrosis factor-α (TNF-α) the supernatants of EC cultures were subjected to differential centrifugation steps to collect larger and smaller EVs which were then characterised by nanoparticle tracking analysis (NTA) and flow cytometry. Mixed with fresh human blood and analysed by thromboelastometry EVs exerted a significant procoagulant stimulus, which could be partly reversed by addition of an anti-TF antibody. Moreover, TF activity was confirmed in the centrifuged fractions. In summary, our results provide evidence of the procoagulant potential of smaller and larger endothelial-derived EV fractions detected by thromboelastometry. The observed effect is most likely due to the release of TF-bearing EVs of different dimensions, which are released upon TNF-α stimulation of endothelial cell cultures. |
| first_indexed | 2024-12-19T04:36:09Z |
| format | Article |
| id | doaj.art-b59d295e2d64436eb0f3caf173c50868 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-19T04:36:09Z |
| publishDate | 2017-06-01 |
| publisher | Nature Portfolio |
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| series | Scientific Reports |
| spelling | doaj.art-b59d295e2d64436eb0f3caf173c508682022-12-21T20:35:44ZengNature PortfolioScientific Reports2045-23222017-06-01711910.1038/s41598-017-03159-0Endothelial Cell-derived Extracellular Vesicles Size-dependently Exert Procoagulant Activity Detected by ThromboelastometryWolfgang Holnthoner0Cornelia Bonstingl1Carina Hromada2Severin Muehleder3Johannes Zipperle4Stefan Stojkovic5Heinz Redl6Johann Wojta7Herbert Schöchl8Johannes Grillari9Sylvia Weilner10Christoph J. Schlimp11Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research CentreLudwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research CentreLudwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research CentreLudwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research CentreLudwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research CentreDepartment of Internal Medicine II, Medical University of ViennaLudwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research CentreDepartment of Internal Medicine II, Medical University of ViennaLudwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research CentreAustrian Cluster for Tissue RegenerationAustrian Cluster for Tissue RegenerationLudwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research CentreAbstract Endothelial cells (ECs) are major modulators of hemostasis by expressing and releasing pro- and anticoagulant mediators into the circulation. Previous studies showed that cultured ECs release procoagulant mediators into cell culture supernatants as evidenced by the reduction of viscoelastic clotting time. This effect was reversed with an anti-tissue factor antibody. Here, we aimed to investigate whether tissue factor (TF) was released by endothelial-derived extracellular vesicles (EVs) and which portion of the released vesicles displays the most prominent procoagulant properties. After stimulation of ECs with tumor-necrosis factor-α (TNF-α) the supernatants of EC cultures were subjected to differential centrifugation steps to collect larger and smaller EVs which were then characterised by nanoparticle tracking analysis (NTA) and flow cytometry. Mixed with fresh human blood and analysed by thromboelastometry EVs exerted a significant procoagulant stimulus, which could be partly reversed by addition of an anti-TF antibody. Moreover, TF activity was confirmed in the centrifuged fractions. In summary, our results provide evidence of the procoagulant potential of smaller and larger endothelial-derived EV fractions detected by thromboelastometry. The observed effect is most likely due to the release of TF-bearing EVs of different dimensions, which are released upon TNF-α stimulation of endothelial cell cultures.https://doi.org/10.1038/s41598-017-03159-0 |
| spellingShingle | Wolfgang Holnthoner Cornelia Bonstingl Carina Hromada Severin Muehleder Johannes Zipperle Stefan Stojkovic Heinz Redl Johann Wojta Herbert Schöchl Johannes Grillari Sylvia Weilner Christoph J. Schlimp Endothelial Cell-derived Extracellular Vesicles Size-dependently Exert Procoagulant Activity Detected by Thromboelastometry Scientific Reports |
| title | Endothelial Cell-derived Extracellular Vesicles Size-dependently Exert Procoagulant Activity Detected by Thromboelastometry |
| title_full | Endothelial Cell-derived Extracellular Vesicles Size-dependently Exert Procoagulant Activity Detected by Thromboelastometry |
| title_fullStr | Endothelial Cell-derived Extracellular Vesicles Size-dependently Exert Procoagulant Activity Detected by Thromboelastometry |
| title_full_unstemmed | Endothelial Cell-derived Extracellular Vesicles Size-dependently Exert Procoagulant Activity Detected by Thromboelastometry |
| title_short | Endothelial Cell-derived Extracellular Vesicles Size-dependently Exert Procoagulant Activity Detected by Thromboelastometry |
| title_sort | endothelial cell derived extracellular vesicles size dependently exert procoagulant activity detected by thromboelastometry |
| url | https://doi.org/10.1038/s41598-017-03159-0 |
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