Role of fibrinogen conformation in platelet activation.
Platelet adhesion and activation induced by fibrinogen (Fbg) coating on polysaccharide layers of hyaluronic acid (Hyal) and its sulfated derivative (HyalS) were analyzed. Hyal or HyalS was coated and grafted on the glass substrate using a photolithographic method. The Fbg coating was achieved by two different routes: the immobilization of Fbg by means of covalent bond to the polysaccharide layers and the mere adsorption of Fbg to Hyal and HyalS surfaces. Platelet adhesion and activation to the surfaces were evaluated using, respectively, scanning electron microscopy (SEM) and quantifying the release of Platelet Factor 4 by ELISA. The method used for the coating of the surfaces with the Fbg influenced the platelet response. In fact, platelet adhesion and activation took place on surfaces covered by bound Fbg but not on those containing adsorbed Fbg. To explain this difference, the molecular mechanism involved in the Fbg--platelet interaction was investigated blocking platelet membrane receptors by monoclonal antibodies. Because the interaction between Fbg and the GPIIb/IIIa platelet membrane receptor was the only molecular pathway involved, Fbg conformation after the interaction (adsorption or binding) with the Hyal and the HyalS chains and the role of serum proteins adsorbed on the Fbg containing surfaces were accurately analyzed. Both adsorbed and bound Fbg prevented the adsorption of further serum proteins; consequently, a direct interaction between Fbg and platelets was supposed and the different platelet behavior was ascribed to the different conformational changes that occurred after the adsorption and the chemical binding of the Fbg to the Hyal and HyalS surfaces.
Chiumiento
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CRISMA and Department of Chemical and Biosystem Sciences and Technologies, University of Siena, Via A. Moro n. 2, Siena 53100, Italy.
Lamponi
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Barbucci
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D000327:Adsorption; D005340:Fibrinogen / Q000737:chemistry* / Q000494:pharmacology; D006820:Hyaluronic Acid; D015539:Platelet Activation / Q000187:drug effects*; D010973:Platelet Adhesiveness; D011134:Polysaccharides; D011487:Protein Conformation; D013499:Surface Properties
D011134:Polysaccharides; D005340:Fibrinogen; D006820:Hyaluronic Acid