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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| Format: | Article |
| Language: | English |
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MDPI AG
2021-01-01
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| Series: | Diagnostics |
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| Online Access: | https://www.mdpi.com/2075-4418/11/2/203 |
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| author | Alexander Dupuy Lejla Hagimola Neil S. A. Mgaieth Callum B. Houlahan Renee E. Preketes-Tardiani Paul R. Coleman Freda H. Passam |
| author_facet | Alexander Dupuy Lejla Hagimola Neil S. A. Mgaieth Callum B. Houlahan Renee E. Preketes-Tardiani Paul R. Coleman Freda H. Passam |
| author_sort | Alexander Dupuy |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-09T03:17:16Z |
| format | Article |
| id | doaj.art-be7e4a3e09b947a6a0c753b45b8be3e1 |
| institution | Directory Open Access Journal |
| issn | 2075-4418 |
| language | English |
| last_indexed | 2024-03-09T03:17:16Z |
| publishDate | 2021-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Diagnostics |
| spelling | doaj.art-be7e4a3e09b947a6a0c753b45b8be3e12023-12-03T15:18:02ZengMDPI AGDiagnostics2075-44182021-01-0111220310.3390/diagnostics11020203Thromboinflammation Model-on-A-Chip by Whole Blood Microfluidics on Fixed Human EndotheliumAlexander Dupuy0Lejla Hagimola1Neil S. A. Mgaieth2Callum B. Houlahan3Renee E. Preketes-Tardiani4Paul R. Coleman5Freda H. Passam6Heart Research Institute, University of Sydney, Sydney, NSW 2042, AustraliaHeart Research Institute, University of Sydney, Sydney, NSW 2042, AustraliaHeart Research Institute, University of Sydney, Sydney, NSW 2042, AustraliaHeart Research Institute, University of Sydney, Sydney, NSW 2042, AustraliaHeart Research Institute, University of Sydney, Sydney, NSW 2042, AustraliaHeart Research Institute, University of Sydney, Sydney, NSW 2042, AustraliaHeart Research Institute, University of Sydney, Sydney, NSW 2042, AustraliaMicrofluidic 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.https://www.mdpi.com/2075-4418/11/2/203neutrophilthromboinflammationmicrofluidicsendotheliumplatelet |
| spellingShingle | Alexander Dupuy Lejla Hagimola Neil S. A. Mgaieth Callum B. Houlahan Renee E. Preketes-Tardiani Paul R. Coleman Freda H. Passam Thromboinflammation Model-on-A-Chip by Whole Blood Microfluidics on Fixed Human Endothelium Diagnostics neutrophil thromboinflammation microfluidics endothelium platelet |
| 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 | neutrophil thromboinflammation microfluidics endothelium platelet |
| url | https://www.mdpi.com/2075-4418/11/2/203 |
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