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 and interference from nonspecifically adsorbed molecules. In this study, we have investigated a technique for covalently immobilizing fibronectin to the PEO-containing triblock copolymer Pluronic 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.
Webb
K
K
W. M. Keck Center for Tissue Engineering, University of Utah, Salt Lake City 84112, USA.
Caldwell
K D
KD
Tresco
P A
PA
D016475:3T3 Cells; D000327:Adsorption; D000818:Animals; D001672:Biocompatible Materials; D002417:Cattle; D002448:Cell Adhesion / Q000502:physiology*; D002455:Cell Division; D002465:Cell Movement; D018914:Cells, Immobilized / Q000502:physiology* / Q000648:ultrastructure; D003599:Cytoskeleton / Q000502:physiology / Q000648:ultrastructure*; D004797:Enzyme-Linked Immunosorbent Assay; D005353:Fibronectins / Q000032:analysis / Q000737:chemistry*; D008297:Male; D051379:Mice; D020442:Poloxamer / Q000737:chemistry*; D011137:Polystyrenes
D001672:Biocompatible Materials; D005353:Fibronectins; D011137:Polystyrenes; D020442:Poloxamer