A novel surfactant-based immobilization method for varying substrate-bound fibronectin

Most biomaterials can be rendered adhesive for anchorage-dependent cells by adsorption of serum, isolated extracellular matrix proteins, or immobilization of peptide sequences. However, difficulties are frequently encountered in characterizing the adsorbed layer due to conformational changes in the molecules following adsorption al td interference from nonspecifically adsorbed molecules. In this study, we have investigated a technique for covalently immobilizing fibronectin to the PEG-containing triblock copolymer Pluronic(TM) F108 ("F108"). We have compared this technique to solution adsorption of fibronectin for its ability to provide controlled variation of bound fibronectin and regulation of fibroblast behavior. Both simple adsorption and covalent immobilization were effective for varying substrate-bound fibronectin. However, adsorption of fibronectin did not effectively regulate fibroblast attachment or spreading in either serum-free or serum-containing media. Fibroblast attachment, spreading, cytoskeletal organization, and proliferation were effectively regulated in response to fibronectin immobilized to F108. Furthermore, F108-treated surfaces without immobilized fibronectin did not support nonspecific fibroblast attachment, even in the presence of serum-containing medium. Fibroblasts were observed to only proliferate on surfaces with high levels of immobilized fibronectin that supported extensive cell spreading and cytoskeletal organization. In summary, covalent immobilization of fibronectin to F108 provided controlled regulation of fibroblast behavior without interference from nonspecific protein adsorption, even in the presence of serum-containing medium. (C) 2000 John Wiley & Sons, Inc.