Quantification of fibrin deposition in flowing blood with peroxidase-labeled fibrinogen. High shear rates induce decreased fibrin deposition and appearance of fibrin monomers
| Autor: | J. J. Sixma, P. N. M. Tijburg, P. G. De Groot, M. J. W. Ijsseldijk |
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| Rok vydání: | 1991 |
| Předmět: |
Endothelium
Enzyme-Linked Immunosorbent Assay Fibrinogen Fibrin Immunoenzyme Techniques Tissue factor medicine Humans Thrombus Cells Cultured Whole blood biology Chemistry Models Cardiovascular Antibodies Monoclonal Anticoagulants Thrombosis medicine.disease Fibrin Monomer Endothelial stem cell medicine.anatomical_structure Biochemistry biology.protein Biophysics Endothelium Vascular Rheology Cardiology and Cardiovascular Medicine medicine.drug |
| Zdroj: | Arteriosclerosis and Thrombosis: A Journal of Vascular Biology. 11:211-220 |
| ISSN: | 1049-8834 |
| DOI: | 10.1161/01.atv.11.2.211 |
| Popis: | To study fibrin incorporation into thrombi at different wall shear rates, a new method to study fibrin deposition on extracellular matrixes underlying stimulated endothelial cells under flow conditions was developed. For this method, we used fibrinogen labeled with peroxidase (Fg-PO). Fg-PO was fully exchangeable for Fg in the clotting assays tested, and PO activity was bound to fibrin-specific fragments. Fg-PO containing fibrin could be stained for microscopic studies with 3,3'-diaminobenzidine and could be quantified by oxidation of phenylenediamine. The absorbance values at 492 nm were converted to fibrin quantities via a standard curve. To study fibrin deposition, Fg-PO was added in trace amounts to whole blood anticoagulated with low-molecular-weight heparin, and perfusion studies were performed over endothelial cell matrixes containing tissue factor. In parallel perfusion studies, 125I-labeled Fg was added in trace amounts to whole blood instead of Fg-PO. Both quantitative methods demonstrated decreased fibrin deposition after perfusions at 1,300 sec-1 compared with fibrin deposition after perfusions at 300 sec-1, while fibrinopeptide A generation was independent of the wall shear rate. The decrease in fibrin deposition at 1,300 sec-1 was accompanied by the appearance of fibrin monomers in the perfusate. This suggested that the decrease in fibrin incorporation at 1,300 sec-1 was due to the impaired polymerization of fibrin monomers. This impairment was probably due to a decrease in local fibrin monomer concentration as a result of the increased removal of monomers from the surface at 1,300 sec-1. |
| Databáze: | OpenAIRE |
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