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Thromboinflammation Model-on-A-Chip by Whole Blood Microfluidics on Fixed Human Endothelium

Microfluidic devices have an established role in the study of platelets and coagulation factors in thrombosis, with potential diagnostic applications. However, few microfluidic devices have assessed the contribution of neutrophils to thrombus formation, despite increasing knowledge of neutrophils’ i...

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Autores principales: Dupuy, Alexander, Hagimola, Lejla, Mgaieth, Neil S. A., Houlahan, Callum B., Preketes-Tardiani, Renee E., Coleman, Paul R., Passam, Freda H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7911484/
https://www.ncbi.nlm.nih.gov/pubmed/33573079
http://dx.doi.org/10.3390/diagnostics11020203
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author Dupuy, Alexander
Hagimola, Lejla
Mgaieth, Neil S. A.
Houlahan, Callum B.
Preketes-Tardiani, Renee E.
Coleman, Paul R.
Passam, Freda H.
author_facet Dupuy, Alexander
Hagimola, Lejla
Mgaieth, Neil S. A.
Houlahan, Callum B.
Preketes-Tardiani, Renee E.
Coleman, Paul R.
Passam, Freda H.
author_sort Dupuy, Alexander
collection PubMed
description Microfluidic devices have an established role in the study of platelets and coagulation factors in thrombosis, with potential diagnostic applications. However, few microfluidic devices have assessed the contribution of neutrophils to thrombus formation, despite increasing knowledge of neutrophils’ importance in cardiovascular thrombosis. We describe a thromboinflammation model which uses straight channels, lined with fixed human umbilical vein endothelial cells, after treatment with tumour necrosis factor-alpha. Re-calcified whole blood is perfused over the endothelium at venous and arterial shear rate. Neutrophil adhesion, platelet and fibrin thrombus formation, is measured over time by the addition of fluorescent antibodies to a whole blood sample. Fixed endothelium retains surface expression of adhesion molecules ICAM-1 and E-Selectin. Neutrophils adhere preferentially to platelet thrombi on the endothelium. Inhibitors of neutrophil adhesion and anti-inflammatory agents, such as isoquercetin, decrease neutrophil adhesion. Our model offers the advantage of the use of (1) fixed endothelium, (2) whole blood, instead of isolated neutrophils, and (3) a small amount of blood (1 mL). The characteristics of this thromboinflammation model provide the potential for further development for drug screening and point-of-care applications.
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spelling pubmed-79114842021-02-28 Thromboinflammation Model-on-A-Chip by Whole Blood Microfluidics on Fixed Human Endothelium Dupuy, Alexander Hagimola, Lejla Mgaieth, Neil S. A. Houlahan, Callum B. Preketes-Tardiani, Renee E. Coleman, Paul R. Passam, Freda H. Diagnostics (Basel) Article Microfluidic devices have an established role in the study of platelets and coagulation factors in thrombosis, with potential diagnostic applications. However, few microfluidic devices have assessed the contribution of neutrophils to thrombus formation, despite increasing knowledge of neutrophils’ importance in cardiovascular thrombosis. We describe a thromboinflammation model which uses straight channels, lined with fixed human umbilical vein endothelial cells, after treatment with tumour necrosis factor-alpha. Re-calcified whole blood is perfused over the endothelium at venous and arterial shear rate. Neutrophil adhesion, platelet and fibrin thrombus formation, is measured over time by the addition of fluorescent antibodies to a whole blood sample. Fixed endothelium retains surface expression of adhesion molecules ICAM-1 and E-Selectin. Neutrophils adhere preferentially to platelet thrombi on the endothelium. Inhibitors of neutrophil adhesion and anti-inflammatory agents, such as isoquercetin, decrease neutrophil adhesion. Our model offers the advantage of the use of (1) fixed endothelium, (2) whole blood, instead of isolated neutrophils, and (3) a small amount of blood (1 mL). The characteristics of this thromboinflammation model provide the potential for further development for drug screening and point-of-care applications. MDPI 2021-01-29 /pmc/articles/PMC7911484/ /pubmed/33573079 http://dx.doi.org/10.3390/diagnostics11020203 Text en © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Dupuy, Alexander
Hagimola, Lejla
Mgaieth, Neil S. A.
Houlahan, Callum B.
Preketes-Tardiani, Renee E.
Coleman, Paul R.
Passam, Freda H.
Thromboinflammation Model-on-A-Chip by Whole Blood Microfluidics on Fixed Human Endothelium
title Thromboinflammation Model-on-A-Chip by Whole Blood Microfluidics on Fixed Human Endothelium
title_full Thromboinflammation Model-on-A-Chip by Whole Blood Microfluidics on Fixed Human Endothelium
title_fullStr Thromboinflammation Model-on-A-Chip by Whole Blood Microfluidics on Fixed Human Endothelium
title_full_unstemmed Thromboinflammation Model-on-A-Chip by Whole Blood Microfluidics on Fixed Human Endothelium
title_short Thromboinflammation Model-on-A-Chip by Whole Blood Microfluidics on Fixed Human Endothelium
title_sort thromboinflammation model-on-a-chip by whole blood microfluidics on fixed human endothelium
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7911484/
https://www.ncbi.nlm.nih.gov/pubmed/33573079
http://dx.doi.org/10.3390/diagnostics11020203
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