Effects of Plasma Proteins on Staphylococcus epidermidis RP62A Adhesion and Interaction with Platelets on Polyurethane Biomaterial Surfaces

Abstract

Plasma proteins influence the initial adhesion of bacteria to biomaterials as well as interactions between bacteria and blood platelets on blood-contacting medical devices. In this paper, we study the effects of three human plasma proteins, albumin, fibrinogen (Fg), and fibronectin (Fn), on the adhesion of Staphylococcus epidemidis RP62A to polyurethane biomaterial surfaces, and also address how these three proteins affect bacterial interactions with human platelets on ma- terials. Measurements of bacterial adhesion on polymer surfaces pre-adsorbed with a variety of proteins demonstrate that Fn leads to increased bacterial adhesion, with the order of effectiveness being Fn Fg > albumin. Immuno-AFM (atomic force microscopy) was used to assess the Fn adsorption/activity on surfaces and bacterial cell membranes by looking at molecular scale events. A correlation between molecular scale Fn adsorption and macroscale bacterial adhe- sion was observed, with an increased numbers of Fn-receptor recognition events measured on cell surfaces as compared to Fg-receptor recognition events, suggesting Fn is an important protein in bacterial adhesion. Monoclonal antibodies recognizing either the carboxyl-terminus or amino-terminus of Fn were coupled to AFM probes and used to assess the orientation of Fn adsorbed on a surface, with an increased amount of Fn carboxyl-terminus availability corresponding to higher bacterial adhesion. Interactions between bacteria and platelets were demonstrated with fluorescence and AFM imaging on the polyurethane surfaces, with albumin inhibiting bacteria-platelet interaction and platelet activation, and both Fg and Fn promoting adhesion of bacteria to platelets and apparent platelet activation, resulting in bacteria/platelet aggregation. 