title: Enhancing the biological activity of immobilized osteopontin using a type-1 collagen affinity coating.
abstract: The covalent attachment of biomolecules onto surfaces represents a step toward the improvement of biomaterial properties by providing relevant biological signals of interest to the cell culture or tissue environment. The chemistries involved, however, often attach proteins to the surface in a random fashion, rather than the conformation or orientation most easily recognized by cells and other proteins both in vitro and in vivo. An alternative approach is to take advantage of natural interactions to both bind and orient a biomolecule "naturally," thereby enhancing its biological activity. Type 1 collagen has been shown to bind to osteopontin (OPN), a protein implicated in processes such as wound healing, endothelial cell survival, and angiogenesis. This study seeks to characterize, quantify, and exploit this interaction in order to present a more naturally recognized form of OPN to the environment surrounding a biomaterial. Binding of OPN to type 1 collagen was confirmed using Surface Plasmon Resonance (SPR). Radio-iodination of OPN showed that binding to collagen was dose-dependent and maximal in basic conditions. Principal component analysis of Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS) data identified differences in OPN immobilized via different techniques. Adhesion of bovine aortic endothelial cells on OPN immobilized using the affinity coating was also significantly enhanced compared to controls. Investigation into the in vivo relevance of this immobilization method is currently underway.
authors.affiliation: Department of Bioengineering, University of Washington, Seattle, Washington 98195, USA.
authors.forename: Stephanie M
authors.initials: SM
authors.lastname: Martin
authors.forename: Jeffrey L
authors.initials: JL
authors.lastname: Schwartz
authors.forename: Cecilia M
authors.initials: CM
authors.lastname: Giachelli
authors.forename: Buddy D
authors.initials: BD
authors.lastname: Ratner
mesh_terms: D000818:Animals; D001639:Bicarbonates / Q000737:chemistry; D002417:Cattle; D002448:Cell Adhesion; D002478:Cells, Cultured; D020099:Coated Materials, Biocompatible / Q000737:chemistry*; D024042:Collagen Type I / Q000737:chemistry*; D042783:Endothelial Cells / Q000378:metabolism; D005079:Excipients; D007093:Imidazoles; D007457:Iodine Radioisotopes; D007553:Isotope Labeling; D013058:Mass Spectrometry; D053495:Osteopontin; D011102:Polyhydroxyethyl Methacrylate; D025341:Principal Component Analysis; D012795:Sialoglycoproteins / Q000008:administration & dosage / Q000032:analysis / Q000494:pharmacology*; D020349:Surface Plasmon Resonance
chemical_list: D001639:Bicarbonates; D020099:Coated Materials, Biocompatible; D024042:Collagen Type I; D005079:Excipients; D007093:Imidazoles; D007457:Iodine Radioisotopes; D012795:Sialoglycoproteins; D053495:Osteopontin; D011102:Polyhydroxyethyl Methacrylate; C006900:N,N-carbonyldiimidazole