Direct quantification of dual protein adsorption dynamics in three dimensional systems in presence of cells.

Abstract

Understanding the composition of the adsorbed protein layer on a biomaterial surface is of an extreme importance as it directs the primary biological response. Direct detection using labeled proteins and indirect detection based on enzymatic assays or changes to mass, refractive index or density of a surface have been so far established. Nevertheless, using current methodologies, detection of multiple proteins simultaneously and particularly in a three-dimensional (3D) substrates is challenging, with the exception of radiolabeling. Here using fluorescence molecular tomography (FMT), we present a non-destructive and versatile approach to quantify adsorption of multiple proteins within 3D environments and reveal the dynamics of adsorption of human serum albumin (HSA) and fibrinogen (Fib) on 3D polymeric scaffold. Furthermore, we show that serum starved human articular chondrocytes in 3D environment preferentially uptake HSA over Fib and to our knowledge this represents the first example of direct visualization and quantification of protein adsorption in a 3D cell culture system.