To investigate the role of neutrophils in thrombus formation, a thromboinflammation model is described that uses straight channels, lined with fixed human umbilical vein endothelial cells, after being treated with tumour necrosis factor-alpha. Re-calcified whole blood is perfused over the endothelium at venous and arterial shear rates. 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. The model offers the advantage of using fixed endothelium and whole blood, instead of isolated neutrophils, and a small amount of blood. The characteristics of this thromboinflammation model provide the potential for further development of drug screening and point-of-care applications.
Thromboinflammation model-on-a-chip by whole blood microfluidics on fixed human endothelium
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