Journal
BIOMATERIALS
Volume 31, Issue 27, Pages 7021-7031Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.biomaterials.2010.05.072
Keywords
Surface modification; Human corneal epithelial cells; Epidermal growth factor (EGF); Coiled-coil interactions; Cell proliferation; Cell adhesion
Funding
- Canada Research Chair on Protein-enhanced Biomaterials
- Natural Sciences and Engineering Research Council of Canada
- Canada Research Chair in Applied Metabolic Engineering
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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. (C) 2010 Elsevier Ltd. All rights reserved.
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