title: Laminin-1 peptide-conjugated chitosan membranes as a novel approach for cell engineering.
abstract: Laminin, a major component of the basement membrane, has diverse biological activities. Recently, we identified various biologically active sequences on laminin-1 by using a large set of synthetic peptides. Chitosan, a polysaccharide, is biodegradable and has been used as a biomaterial. Here, we conjugated several biologically active laminin peptides onto chitosan membranes and measured the cell attachment activity of peptide-conjugated chitosan membranes with various cell types. The active laminin peptide-conjugated chitosan membranes promoted cell attachment with cell type specificity. A99 (AGTFALRGDNPQG)-chitosan membrane promoted cell attachment with well-organized actin stress fibers. This adhesion was inhibited by EDTA but not by heparin. AG73 (RKRLQVQLSIRT)-chitosan membrane promoted cell attachment with filopodia formation, and this adhesion was inhibited by heparin but not by EDTA. These data suggest that the A99-chitosan membrane interacted with an integrin cellular receptor and that the AG73-chitosan membrane promoted proteoglycan-mediated cell attachment, as previously reported. Furthermore, both AG73-chitosan and A99-chitosan membranes effectively promoted neurite outgrowth with PC12 rat pheochromocytoma cells. We conclude that conjugation on a chitosan membrane is applicable for testing quantitatively the biological activity of synthetic peptides and that these constructs have a potential ability to serve as bioadhesive materials for tissue regeneration and engineering.
authors.affiliation: Laboratory of Bio-Material Chemistry, Division of Bioscience, Graduate School of Environmental Earth Science Hokkaido University, Kita 10 Nishi 5, Kita-ku, Sapporo 060-0810, Japan. nomizu@ees.hokudai.ac.jp
authors.forename: Mayumi
authors.initials: M
authors.lastname: Mochizuki
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Kadoya
authors.forename: Yoko
authors.initials: Y
authors.lastname: Wakabayashi
authors.forename: Kozue
authors.initials: K
authors.lastname: Kato
authors.forename: Ikuko
authors.initials: I
authors.lastname: Okazaki
authors.forename: Masanori
authors.initials: M
authors.lastname: Yamada
authors.forename: Taku
authors.initials: T
authors.lastname: Sato
authors.forename: Nobuo
authors.initials: N
authors.lastname: Sakairi
authors.forename: Norio
authors.initials: N
authors.lastname: Nishi
authors.forename: Motoyoshi
authors.initials: M
authors.lastname: Nomizu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods; D002455:Cell Division / Q000502:physiology; D002686:Chitin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism*; D048271:Chitosan; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D007797:Laminin / Q000737:chemistry*; D008567:Membranes, Artificial; D016716:PC12 Cells; D051381:Rats
chemical_list: D007797:Laminin; D008567:Membranes, Artificial; C121346:laminin 1; D002686:Chitin; D048271:Chitosan