title: Nanofibrillar hydrogel scaffolds from recombinant protein-based polymers with integrin- and proteoglycan-binding domains.
abstract: This study describes the design, production, and testing of functionalized variants of a recombinant protein-based polymer that forms nanofibrillar hydrogels with self-healing properties. With a view to bone tissue engineering applications, we equipped these variants with N-terminal extensions containing either (1) integrin-binding (RGD) or (2) less commonly studied proteoglycan-binding (KRSR) cell-adhesive motifs. The polymers were efficiently produced as secreted proteins using the yeast Pichia pastoris and were essentially monodisperse. The pH-responsive protein-based polymers are soluble at low pH and self-assemble into supramolecular fibrils and hydrogels at physiological pH. By mixing functionalized and nonfunctionalized proteins in different ratios, and adjusting pH, hydrogel scaffolds with the same protein concentration but varying content of the two types of cell-adhesive motifs were readily obtained. The scaffolds were used for the two-dimensional culture of MG-63 osteoblastic cells. RGD domains had a slightly stronger effect than KRSR domains on adhesion, activity, and spreading. However, scaffolds featuring both functional domains revealed a clear synergistic effect on cell metabolic activity and spreading, and provided the highest final degree of cell confluency. The mixed functionalized hydrogels presented here thus allowed to tailor the osteoblastic cell response, offering prospects for their further development as scaffolds for bone regeneration. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 3082-3092, 2016.
authors.affiliation: Physical Chemistry and Soft Matter, Wageningen University, Stippeneng 4, NL-6708 WE Wageningen, The Netherlands.
authors.forename: Małgorzata K
authors.initials: MK
authors.lastname: Włodarczyk-Biegun
authors.affiliation: Wageningen UR Food & Biobased Research, Bornse Weilanden 9, NL-6708 WG Wageningen, The Netherlands. marc.werten@wur.nl.
authors.forename: Marc W T
authors.initials: MW
authors.lastname: Werten
authors.affiliation: Physical Chemistry and Soft Matter, Wageningen University, Stippeneng 4, NL-6708 WE Wageningen, The Netherlands.|Faculty of Materials Science and Ceramics, Department of Biomaterials, AGH University of Science and Technology, Al. A. Mickiewicza 30, Krakow, 30-059, Poland.
authors.forename: Urszula
authors.initials: U
authors.lastname: Posadowska
authors.affiliation: Physical Chemistry and Soft Matter, Wageningen University, Stippeneng 4, NL-6708 WE Wageningen, The Netherlands.
authors.forename: Ingeborg M
authors.initials: IM
authors.lastname: Storm
authors.affiliation: Wageningen UR Food & Biobased Research, Bornse Weilanden 9, NL-6708 WG Wageningen, The Netherlands.
authors.forename: Frits A
authors.initials: FA
authors.lastname: de Wolf
authors.affiliation: Department of Biomaterials, Radboudumc, Philips Van Leydenlaan 25, NL-6525 EX Nijmegen, The Netherlands.
authors.forename: Jeroen J J P
authors.initials: JJ
authors.lastname: van den Beucken
authors.affiliation: Department of Biomaterials, Radboudumc, Philips Van Leydenlaan 25, NL-6525 EX Nijmegen, The Netherlands.
authors.forename: Sander C G
authors.initials: SC
authors.lastname: Leeuwenburgh
authors.affiliation: Physical Chemistry and Soft Matter, Wageningen University, Stippeneng 4, NL-6708 WE Wageningen, The Netherlands.
authors.forename: Martien A
authors.initials: MA
authors.lastname: Cohen Stuart
authors.affiliation: Physical Chemistry and Soft Matter, Wageningen University, Stippeneng 4, NL-6708 WE Wageningen, The Netherlands.
authors.forename: Marleen
authors.initials: M
authors.lastname: Kamperman
mesh_terms: D001665:Binding Sites; D002448:Cell Adhesion; D002460:Cell Line; D002470:Cell Survival; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry*; D016023:Integrins / Q000378:metabolism*; D057139:Nanofibers / Q000737:chemistry* / Q000648:ultrastructure; D009842:Oligopeptides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism; D010843:Pichia / Q000235:genetics; D011485:Protein Binding; D011509:Proteoglycans / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D023822:Tissue Engineering; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D016023:Integrins; D009842:Oligopeptides; D011509:Proteoglycans; D011994:Recombinant Proteins; C437191:lysyl-arginyl-seryl-arginine; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; C047981:arginyl-glycyl-aspartic acid
keywords: KRSR cell-adhesive domain; Pichia pastoris; protein-based polymers; self-assembled hydrogel; tissue engineering