Time-Dependent Conformational Changes in Adsorbed Albumin and Its Effect on Platelet Adhesion

Abstract Recent studies have shown that platelets can adhere to adsorbed albumin (Alb) through a receptor-mediated mechanism, but only if the Alb undergoes more than a critical degree of adsorption-induced unfolding. The objectives of this research were to investigate whether Alb that wasinitially adsorbed in a manner that induced unfolding that was less than this critical level wouldundergo further unfolding with time; and if so, whether this would induce the onset of plateletadhesion once this critical level was exceeded. To address these questions, CD spectropolarimetrywas used to monitor the structure of Alb on OH- and CH 3 -functionalized alkanethiol self-assembled monolayer surfaces, with the Alb initially adsorbed under conditions resulting indegrees of unfolding that were below this critical level, and then the adsorbed Alb layers wereaged over a six-month period of time in sterile physiological saline at 37°C. Platelet adhesion toAlb was quantified at selected time points via a lactate dehydrogenase (LDH) assay. The resultsindicate that an adsorbed Alb layer does undergo further structural changes with increasingresidence time and supports platelet adhesion once it unfolds beyond the previously determinedcritical level. These results may be relevant to the clinically observed problem of the onset of late-thrombosis, which occurs on cardiovascular implants such as drug-eluting stents.