title: Mobility of the Arg-Gly-Asp ligand on the outermost surface of biomaterials suppresses integrin-mediated mechanotransduction and subsequent cell functions.
abstract: Mechanotransduction in the regulation of cellular responses has been previously studied using elastic hydrogels. Because cells interact only with the surface of biomaterials, we are focusing on the molecular mobility at the outermost surface of biomaterials. In this study, surfaces with the mobile Arg-Gly-Asp-Ser (RGDS) peptide have been constructed. Cell culture substrates were coated with ABA-type block copolymers composed of poly(2-methacryloyloxyethyl phosphorylcholine-co-n-butyl methacrylate) segments (A) and a polyrotaxane (PRX) unit with RGDS bound to α-cyclodextrin (B). Adhesion, morphological changes and actin filament formation of human umbilical vein endothelial cells were reduced on the surfaces containing mobile PRX-RGDS in comparison to the immobile RGDS surfaces constructed from random copolymers with RGDS side groups (Prop-andom-RGDS). In the neurite outgrowth assay using rat adrenal pheochromocytoma cells (PC12), only ∼20% of adherent PC12 cells had neurites on PRX-RGDS surfaces, but more than 50% did on the Random-RGDS surface. The beating colony of dimethyl-sulfoxide-treated mouse embryonic carcinoma cells (P19CL6) were found 10 and 14 days after induction on PRX-RGDS and Random-RGDS surfaces, respectively. After 22 days, the beating colony disappeared on PRX-RGDS surfaces, but many colonies remained on Random-RGDS surfaces. These data suggest that the molecular mobility of the cell-binding ligand on the outermost surface of materials effectively suppresses the actin filament formation and differentiation of these functional cell lines, and may be used as a culture substrate for immature stem cells or progenitor cells.
authors.affiliation: Department of Biomedical Engineering, National Cerebral and Cardiovascular Center Research Institute, 5-7-1 Fujishirodai, Suita, Osaka 565-8565, Japan; JST-CREST, 7 Gobancho, Chiyoda-ku, Tokyo 102-0076, Japan.
authors.forename: Sachiro
authors.initials: S
authors.lastname: Kakinoki
authors.affiliation: Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10, Kanda-Surugadai, Chiyoda-ku, Tokyo 101-0062, Japan; JST-CREST, 7 Gobancho, Chiyoda-ku, Tokyo 102-0076, Japan.
authors.forename: Ji-Hun
authors.initials: JH
authors.lastname: Seo
authors.affiliation: Department of Materials Engineering, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan; JST-CREST, 7 Gobancho, Chiyoda-ku, Tokyo 102-0076, Japan.
authors.forename: Yuuki
authors.initials: Y
authors.lastname: Inoue
authors.affiliation: Department of Materials Engineering, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan; JST-CREST, 7 Gobancho, Chiyoda-ku, Tokyo 102-0076, Japan.
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Ishihara
authors.affiliation: Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10, Kanda-Surugadai, Chiyoda-ku, Tokyo 101-0062, Japan; JST-CREST, 7 Gobancho, Chiyoda-ku, Tokyo 102-0076, Japan.
authors.forename: Nobuhiko
authors.initials: N
authors.lastname: Yui
authors.affiliation: Department of Biomedical Engineering, National Cerebral and Cardiovascular Center Research Institute, 5-7-1 Fujishirodai, Suita, Osaka 565-8565, Japan; JST-CREST, 7 Gobancho, Chiyoda-ku, Tokyo 102-0076, Japan. Electronic address: yamtet@ncvc.go.jp.
authors.forename: Tetsuji
authors.initials: T
authors.lastname: Yamaoka
mesh_terms: D008841:Actin Cytoskeleton; D000818:Animals; D002454:Cell Differentiation; D002465:Cell Movement; D003505:Cyclodextrins / Q000737:chemistry; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D040542:Mechanotransduction, Cellular; D051379:Mice; D009842:Oligopeptides / Q000737:chemistry*; D016716:PC12 Cells; D020442:Poloxamer / Q000737:chemistry; D011109:Polymethacrylic Acids / Q000737:chemistry; D051381:Rats; D043862:Rotaxanes / Q000737:chemistry; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*; D047391:alpha-Cyclodextrins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D003505:Cyclodextrins; D009842:Oligopeptides; D011109:Polymethacrylic Acids; D043862:Rotaxanes; D047391:alpha-Cyclodextrins; C475673:polyrotaxane; D020442:Poloxamer; C046617:arginyl-glycyl-aspartyl-serine
keywords: Cell adhesion; Cell differentiation; Molecular mobility; Polyrotaxane; RGDS