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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. STATEMENT OF SIGNIFICANCE:The biomaterial surface upon exposure to biological fluids is covered by a layer of proteins, which is modified over a period of time and dictates the fate of the biomaterial. In this study, we present and validate a new methodology for quantification of protein adsorption on to a three-dimensional polymer scaffold from unitary and binary systems, using fluorescence molecular tomography, an optical trans-illumination technique with picomolar sensitivity. In additional to being able to follow behavior of two proteins simultaneously, this methodology is also suitable for studying protein uptake in cells situated in a polymer environment. The ability to follow protein adsorption/uptake in a continuous manner opens up new possibilities to study the role of serum proteins in biomaterial compatibility.

journal_name

Acta Biomater

journal_title

Acta biomaterialia

authors

Sarem M,Vonwil D,Lüdeke S,Shastri VP

doi

10.1016/j.actbio.2017.05.021

subject

Has Abstract

pub_date

2017-07-15 00:00:00

pages

285-292

eissn

1742-7061

issn

1878-7568

pii

S1742-7061(17)30301-X

journal_volume

pub_type

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