title: A highly versatile adaptor protein for the tethering of growth factors to gelatin-based biomaterials.
abstract: In the field of tissue engineering, the tethering of growth factors to tissue scaffolds in an oriented manner can enhance their activity and increase their half-life. We chose to investigate the capture of the basic Fibroblast Growth Factor (bFGF) and the Epidermal Growth Factor (EGF) on a gelatin layer, as a model for the functionalization of collagen-based biomaterials. Our strategy relies on the use of two high affinity interactions, that is, the one between two distinct coil peptides as well as the one occurring between a collagen-binding domain (CBD) and gelatin. We expressed a chimeric protein to be used as an adaptor that comprises one of the coil peptides and a CBD derived from the human fibronectin. We proved that it has the ability to bind simultaneously to a gelatin substrate and to form a heterodimeric coiled-coil domain with recombinant growth factors being tagged with the complementary coil peptide. The tethering of the growth factors was characterized by ELISA and surface plasmon resonance-based biosensing. The bioactivity of the immobilized bFGF and EGF was evaluated by a human umbilical vein endothelial cell proliferation assay and a vascular smooth muscle cell survival assay. We found that the tethering of EGF preserved its mitogenic and anti-apoptotic activity. In the case of bFGF, when captured via our adaptor protein, changes in its natural mode of interaction with gelatin were observed.
  
  
  
  In an effort to functionalize collagen/gelatin-based biomaterials with growth factors, we have designed an adaptor protein corresponding to a collagen-binding domain fused to a coil peptide. In our strategy, this adaptor protein captures growth factors being tagged with the partner coil peptide in a specific, stable and oriented manner. We have found that the tethering of the Epidermal Growth Factor preserved its mitogenic and anti-apoptotic activity. In the case of the basic Fibroblast Growth Factor, the captured growth factor remained bioactive although its tethering via this adaptor protein modified its natural mode of interaction with gelatin. Altogether this strategy is easily adaptable to the simultaneous tethering of various growth factors.
authors.affiliation: Department of Chemical Engineering, Biomedical Science and Technology Research Group, Bio-P(2) Research Unit, École Polytechnique de Montréal, P.O. Box 6079, succ. Centre-Ville, Montréal (QC) H3C 3A7, Canada.
authors.forename: Cyril
authors.initials: C
authors.lastname: Addi
authors.affiliation: Department of Chemical Engineering, Biomedical Science and Technology Research Group, Bio-P(2) Research Unit, École Polytechnique de Montréal, P.O. Box 6079, succ. Centre-Ville, Montréal (QC) H3C 3A7, Canada.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Murschel
authors.affiliation: Department of Chemical Engineering, Biomedical Science and Technology Research Group, Bio-P(2) Research Unit, École Polytechnique de Montréal, P.O. Box 6079, succ. Centre-Ville, Montréal (QC) H3C 3A7, Canada.
authors.forename: Benoît
authors.initials: B
authors.lastname: Liberelle
authors.affiliation: Department of Chemical Engineering, Biomedical Science and Technology Research Group, Bio-P(2) Research Unit, École Polytechnique de Montréal, P.O. Box 6079, succ. Centre-Ville, Montréal (QC) H3C 3A7, Canada.
authors.forename: Nesrine
authors.initials: N
authors.lastname: Riahi
authors.affiliation: Department of Chemical Engineering, Biomedical Science and Technology Research Group, Bio-P(2) Research Unit, École Polytechnique de Montréal, P.O. Box 6079, succ. Centre-Ville, Montréal (QC) H3C 3A7, Canada. Electronic address: gregory.decrescenzo@polymtl.ca.
authors.forename: Gregory
authors.initials: G
authors.lastname: De Crescenzo
mesh_terms: D001672:Biocompatible Materials / Q000737:chemistry / Q000494:pharmacology; D004815:Epidermal Growth Factor / Q000737:chemistry / Q000494:pharmacology; D016222:Fibroblast Growth Factor 2 / Q000737:chemistry / Q000494:pharmacology; D005353:Fibronectins / Q000737:chemistry / Q000494:pharmacology; D005780:Gelatin / Q000737:chemistry / Q000494:pharmacology; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry / Q000494:pharmacology; D008422:Materials Testing / Q000379:methods; D000072417:Protein Domains; D011993:Recombinant Fusion Proteins / Q000494:pharmacology; D023822:Tissue Engineering / Q000379:methods
chemical_list: D001672:Biocompatible Materials; D005353:Fibronectins; D055767:Immobilized Proteins; D011993:Recombinant Fusion Proteins; D016222:Fibroblast Growth Factor 2; D004815:Epidermal Growth Factor; D005780:Gelatin
keywords: Biofunctionalization; Coiled-coil; Collagen-binding domain; Growth factor; Tethering