Fibronectin immobilized by a novel surface treatment regulates fibroblast attachment and spreading

In order to understand the influence of cell-adhesive molecules on anchorage-dependent cell behavior on biomaterial surfaces, a model system is required where these molecules can be applied to surfaces with controlled surface ligand density and resistance to the adsorption of additional proteins present in the medium. This study asked whether fibronectin could be immobilized in a controlled manner to a hydrophobic surface with a chemically modified triblock surfactant. ELISA studies indicated that variation of the soluble fibronectin concentration used for immobilization could be used to control the amount of fibronectin immobilized to the surface. Furthermore, fibroblasts seeded on these surfaces in 10% serum-containing medium attached and spread as a function of the amount of immobilized fibronectin. Surfaces treated with unmodified surfactant did not support cell attachment, suggesting that cell attachment and spreading were primarily regulated by the immobilized fibronectin with minimal interference from adsorption of serum proteins. Together, these results suggest that covalent immobilization to Pluronic F108 provides a method for studying cellular responses to cell adhesive proteins with little interference from competing adsorbates, even in the presence of complex biological fluids such as serum. This technique may be applicable to a variety of existing hydrophobic biomedical polymers as a basic science tool as well as for influencing cell behavior at implant interfaces.