title: Human corneal epithelial cell response to epidermal growth factor tethered via coiled-coil interactions.
abstract: The development of new strategies for protein immobilization to control cell adhesion, growth and differentiation is of prime interest in the field of tissue engineering. Here we propose a versatile approach based on the interaction between two de novo designed peptides, Ecoil and Kcoil, for oriented immobilization of epidermal growth factor (EGF) on polyethylene terephthalate (PET) films. After amination of PET surfaces by ammonia plasma treatment, Kcoil peptides were covalently grafted in an oriented fashion using succinimidyl 6-[30-(2-pyridyldithio)-propionamido] hexanoate (LC-SPDP) linker, and the Kcoil-functionalized films were characterized by X-ray photoelectron spectroscopy (XPS). Bioactivity of Ecoil-EGF captured on Kcoil-functionalized PET via coiled-coil interactions was confirmed by EGF receptor phosphorylation analysis following A-431 cell attachment. We also demonstrated cell biological effects where tethered EGF enhanced adhesion, spreading and proliferation of human corneal epithelial cells compared to EGF that was either physically adsorbed or present in solution. Tethered EGF effects were most likely linked to the prolonged activation of both mitogen-activated protein kinase and phosphoinositidine 3-kinase pathways. Taken together, our results indicate that coiled-coil-based oriented immobilization is a powerful method to specifically tailor biomaterial surfaces for tissue engineering applications.
authors.affiliation: Department of Chemical Engineering, Groupe de Recherche en Sciences et Technologie Biomédicales, Bio-P(2) Research Unit, Ecole Polytechnique de Montréal, succ. Centre-Ville, Montréal, QC, Canada.
authors.forename: Cyril
authors.initials: C
authors.lastname: Boucher
authors.forename: Juan-Carlos
authors.initials: JC
authors.lastname: Ruiz
authors.forename: Marc
authors.initials: M
authors.lastname: Thibault
authors.forename: Michael D
authors.initials: MD
authors.lastname: Buschmann
authors.forename: Michael R
authors.initials: MR
authors.lastname: Wertheimer
authors.forename: Mario
authors.initials: M
authors.lastname: Jolicoeur
authors.forename: Yves
authors.initials: Y
authors.lastname: Durocher
authors.forename: Gregory
authors.initials: G
authors.lastname: De Crescenzo
mesh_terms: D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D003315:Cornea / Q000166:cytology*; D004815:Epidermal Growth Factor / Q000737:chemistry / Q000494:pharmacology*; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D066246:ErbB Receptors / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D018613:Microscopy, Confocal; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D056951:Photoelectron Spectroscopy; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D004815:Epidermal Growth Factor; D019869:Phosphatidylinositol 3-Kinases; D066246:ErbB Receptors; D020928:Mitogen-Activated Protein Kinases