The study of endothelial dysfunction (ED) is crucial to identify the pathogenetic mecha- nism(s) and provide indications for patient management in cardiovascular diseases. It is currently hindered by the limited availability of patient-specific primary endothelial cells (ECs). Endothe- lial colony-forming cells (ECFCs) represent an optimal non-invasive tool to overcome this issue. Therefore, we investigated the use of ECFCs as a substrate in thrombogenesis and thrombin genera- tion assay (TGA) to assess ED. Both assays were set up on human umbilical vein endothelial cells (HUVECs) and then tested on ECFCs obtained from healthy donors. To prove the ability of the assays to detect endothelial activation, ECs stimulated with TNFα were compared with unstimulated ECs. EC activation was confirmed by the upregulation of VCAM-1 and Tissue Factor expression. Both assays discriminated between unstimulated and activated HUVECs and ECFCs, as significantly higher platelet deposition and fibrin formation in thrombogenesis assay, and thrombin generation in TGA, were observed when TNFα-activated ECs were used as a substrate. The amount of fibrin and thrombin measured in the two assays were directly correlated. Our results support the combined use of a thrombogenesis assay and TGA performed on patient-derived ECFCs to provide a personalized global assessment of ED relevant to the patient’s hemostatic profile.

Development of personalized thrombogenesis and thrombin generation assays to assess endothelial dysfunction in cardiovascular diseases / M. Bacci, A. Cancellara, R. Ciceri, E. Romualdi, V. Pessi, F. Tumminello, M. Fantuzzi, M. Donadini, C. Lodigiani, S. Della Bella, F. Calcaterra, D. Mavilio. - In: BIOMEDICINES. - ISSN 2227-9059. - 11:6(2023 Jun 08), pp. 1669.1-1669.13. [10.3390/biomedicines11061669]

Development of personalized thrombogenesis and thrombin generation assays to assess endothelial dysfunction in cardiovascular diseases

A. Cancellara
Co-primo
;
R. Ciceri;S. Della Bella
;
F. Calcaterra
Co-ultimo
;
D. Mavilio
Co-ultimo
2023

Abstract

The study of endothelial dysfunction (ED) is crucial to identify the pathogenetic mecha- nism(s) and provide indications for patient management in cardiovascular diseases. It is currently hindered by the limited availability of patient-specific primary endothelial cells (ECs). Endothe- lial colony-forming cells (ECFCs) represent an optimal non-invasive tool to overcome this issue. Therefore, we investigated the use of ECFCs as a substrate in thrombogenesis and thrombin genera- tion assay (TGA) to assess ED. Both assays were set up on human umbilical vein endothelial cells (HUVECs) and then tested on ECFCs obtained from healthy donors. To prove the ability of the assays to detect endothelial activation, ECs stimulated with TNFα were compared with unstimulated ECs. EC activation was confirmed by the upregulation of VCAM-1 and Tissue Factor expression. Both assays discriminated between unstimulated and activated HUVECs and ECFCs, as significantly higher platelet deposition and fibrin formation in thrombogenesis assay, and thrombin generation in TGA, were observed when TNFα-activated ECs were used as a substrate. The amount of fibrin and thrombin measured in the two assays were directly correlated. Our results support the combined use of a thrombogenesis assay and TGA performed on patient-derived ECFCs to provide a personalized global assessment of ED relevant to the patient’s hemostatic profile.
endothelial dysfunction; endothelial colony-forming cells (ECFCs); thrombogenesis assay; thrombin generation assay (TGA);
Settore MED/46 - Scienze Tecniche di Medicina di Laboratorio
Settore MED/04 - Patologia Generale
Settore MED/15 - Malattie del Sangue
8-giu-2023
Article (author)
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/975368
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