Polymer chain flexibility-induced differences in fetuin A adsorption and its implications on cell attachment and proliferation

Tissue cells are known to respond to the stiffness of the polymer substrate on which they are grown. It has been suggested that material stiffness influences the composition of the protein layer that adsorbs to the material surface, which affects subsequent cell behavior. Previously, the stiffness of a biomaterial elastomer formed from an acrylated star-poly(d,l lactide-co-ε-caprolactone) was found to influence both fibroblast proliferation as well as the adsorption of certain proteins. However, it remained unresolved as to whether material stiffness influenced protein adsorption from serum supplemented environments and which protein(s) may have been responsible for the difference in fibroblast proliferation. Using quantitative proteomics, we show that polymer stiffness influenced the composition of the protein layers that adsorb from serum supplemented media. Fetuin A was identified as a protein that influenced fibroblast proliferation and, when combined with basic fibroblast growth factor as a medium supplement, improved fibroblast proliferation over 14 days. This study is the first to correlate cell proliferation to surface adsorbed fetuin A and presents the potential new application for fetuin A as biomaterial coating or surface modifier. This work also demonstrates a novel application of quantitative proteomics for the investigation of competitive protein adsorption to biomaterial surfaces.Statement of significanceCells are able to respond to the stiffness of their material substrate, but the method by which they sense material stiffness is still under investigation. Previously, material stiffness was found to impact the individual adsorption of fibronectin, a protein associated with cell attachment; however, it was unclear if stiffness was able to affect protein adsorption in environments with multiple proteins. This study shows that material stiffness affects the compositions of protein layers adsorbed from supplemented media, and suggests that cells may sense material stiffness via the adsorbed protein layer. Interestingly, fetuin A was found to be affecting cell proliferation and not fibronectin. Finally, this research demonstrates the use of relative quantitation proteomics as a potentially powerful method to improve biomaterial compatibility.

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