title: Protein and platelet interactions with thermally denatured fibrinogen and cross-linked fibrin coated surfaces.
abstract: In this work the hypothesis that a mature, cross-linked fibrin clot, pre-formed on a biomaterial, may be relatively nonthrombogenic was investigated. A cross-linked fibrin layer was formed on polyethylene which had been precoated with thermally denatured fibrinogen. Plasma protein adsorption and platelet interactions with the cross-linked fibrin and denatured fibrinogen surfaces were investigated. The adsorption of albumin, fibrinogen, and fibronectin from plasma was measured. For all three proteins, the cross-linked fibrin surface exhibited much higher levels of adsorption than either the thermally denatured fibrinogen or the polyethylene surface. Vroman peaks were observed for fibrinogen and fibronectin on polyethylene but not on the cross-linked fibrin and thermally denatured fibrinogen materials. In dilute plasma the thermally denatured fibrinogen surface showed considerable resistance to protein adsorption. However, at plasma concentrations greater than about 5% normal, this protein resistance was apparently lost. Platelet interactions (adhesion and release of granule constituents from adherent platelets) using suspensions of washed platelets in the presence of red cells were investigated at shear rates of 50, 300, and 525 s(-1) using a cone and plate apparatus. The levels of platelet adhesion on the different surfaces were in the order: adsorbed fibrinogen > cross-linked fibrin > thermally denatured fibrinogen = polyethylene. Platelets on the cross-linked fibrin surface also showed high levels of release indicating significant platelet activation. Scanning electron microscopic observations were in agreement with the platelet adhesion and release data, showing only a few (but well-spread) adherent platelets on the cross-linked fibrin surface.
authors.affiliation: Department of Chemical Engineering, McMaster University, Hamilton, Ontario, Canada.
authors.forename: G A
authors.initials: GA
authors.lastname: Skarja
authors.forename: J L
authors.initials: JL
authors.lastname: Brash
authors.forename: P
authors.initials: P
authors.lastname: Bishop
authors.forename: K A
authors.initials: KA
authors.lastname: Woodhouse
mesh_terms: D000327:Adsorption; D001792:Blood Platelets / Q000166:cytology* / Q000648:ultrastructure; D020099:Coated Materials, Biocompatible / Q000737:chemistry*; D003432:Cross-Linking Reagents / Q000737:chemistry; D015718:Fibrin Tissue Adhesive / Q000737:chemistry*; D005340:Fibrinogen / Q000737:chemistry* / Q000648:ultrastructure; D006358:Hot Temperature; D006801:Humans; D008855:Microscopy, Electron, Scanning; D010973:Platelet Adhesiveness; D011095:Polyethylenes / Q000737:chemistry; D011489:Protein Denaturation
chemical_list: D020099:Coated Materials, Biocompatible; D003432:Cross-Linking Reagents; D015718:Fibrin Tissue Adhesive; D011095:Polyethylenes; D005340:Fibrinogen