Human corneal epithelial cell response to epidermal growth factor tethered via coiled-coil interactions

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DOIResolve DOI: http://doi.org/10.1016/j.biomaterials.2010.05.072
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TypeArticle
Journal titleBiomaterials
Volume31
Issue27
Pages70217031; # of pages: 11
SubjectAmmonia; Analysis; Bio; Cell adhesion; Cells; Epidermal growth factor; Epithelial cells; Human; Peptides; Phosphorylation; Protein; Tissue engineering; X-ray; Surface modification; Human corneal epithelial cells; Coiled-coil interaction; Cell proliferation; Cell adhesion
AbstractThe 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
Publication date
LanguageEnglish
AffiliationNational Research Council Canada (NRC-CNRC); NRC Biotechnology Research Institute
Peer reviewedYes
NRC number52760
NPARC number16225330
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Record identifier1c871e5a-994a-4e4f-a265-8e5e4a5cad1d
Record created2010-11-05
Record modified2016-05-09
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