Platelet adhesion to radiofrequency glow-discharge-deposited fluorocarbon polymers preadsorbed with selectively depleted plasmas show the primary role of fibrinogen.
Fluorocarbon radio-frequency glow-discharge (RFGD) treatment has previously been shown to cause decreased platelet adhesion despite the presence of adsorbed fibrinogen on the surfaces. In this study platelet adhesion to fluorocarbon RFGD-treated surfaces preadsorbed with human plasma was further examined. A series of plasma deposited fluorocarbon thin films were made by varying the C3F6/CH4 ratio in the monomer feed. The surfaces were preadsorbed with plasma, serum, or plasma selectively depleted of fibronectin, vitronectin, or Von Willebrand factor, and platelet adhesion was measured. We also measured fibrinogen adsorption to the surfaces from plasma, monoclonal antibody binding to adsorbed fibrinogen and SDS elutability of the adsorbed fibrinogen. The antibodies used bind to the three putative platelet binding sites on fibrinogen, namely, M1 antibody binds to the dodecapeptide at the C-terminus of the gamma chain, gamma (402-411), R1 antibody binds to a sequence in the Aalpha chain (87-100) which includes RGDF at Aalpha (95-98) and R2 antibody binds a sequence in the Aalpha chain (566-580) which includes RGDS at Aalpha (572-575). Fibrinogen was found to play a decisive role in mediating platelet adhesion to the fluorocarbon surfaces contacting plasma. Few platelets adhered to the fluorocarbon surfaces preadsorbed with serum, while preadsorption with plasma selectively-depleted of either fibronectin, vitronectin, or von Willebrand factor did not decrease platelet adhesion significantly. Replenishment of exogenous fibrinogen to serum restored platelet adhesion, while replenishment of the other proteins had no effect. Platelet adhesion to the fluorocarbon surfaces was lower than to PET or the methane glow-discharge-treated PET. However, there was no apparent correlation between platelet adhesion and the amount of fibrinogen adsorption or monoclonal antibody binding to surface-bound fibrinogen.
Department of Bioengineering, University of Washington, Seattle, WA 98195, USA.
W B
WB
Tsai
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Shi
J M
JM
Grunkemeier
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C
McFarland
T A
TA
Horbett
D000327:Adsorption; D000911:Antibodies, Monoclonal / Q000737:chemistry; D001672:Biocompatible Materials; D001798:Blood Proteins / Q000737:chemistry; D002021:Buffers; D002448:Cell Adhesion; D004797:Enzyme-Linked Immunosorbent Assay; D005340:Fibrinogen / Q000737:chemistry / Q000502:physiology*; D005353:Fibronectins / Q000737:chemistry; D005466:Fluorocarbons / Q000737:chemistry*; D006801:Humans; D010973:Platelet Adhesiveness; D011093:Polyethylene Terephthalates / Q000737:chemistry; D011108:Polymers / Q000737:chemistry*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D013499:Surface Properties; D013997:Time Factors; D019096:Vitronectin / Q000737:chemistry; D014867:Water; D014841:von Willebrand Factor / Q000737:chemistry
D000911:Antibodies, Monoclonal; D001672:Biocompatible Materials; D001798:Blood Proteins; D002021:Buffers; D005353:Fibronectins; D005466:Fluorocarbons; D011093:Polyethylene Terephthalates; D011108:Polymers; D019096:Vitronectin; D014841:von Willebrand Factor; D014867:Water; D012967:Sodium Dodecyl Sulfate; D005340:Fibrinogen