title: Cell surface receptor-specific scaffold requirements for adhesion to laminin-derived peptide-chitosan membranes.
abstract: Scaffolds are used for bioengineering to regulate cellular functions. Previously, we developed laminin-derived peptide-conjugated chitosan membranes for cell engineering. Here, we determined whether changes in the chitosan scaffold altered the cellular response. When an alphavbeta3 integrin-binding peptide A99a (ALRGDN) was conjugated on chitosan membranes of varying density (1.5-1500 ng/mm(2)), cell adhesion was altered depending on the amount of chitosan. 3 or 30 ng/mm(2) of the A99a-chitosan membrane effectively promoted cell attachment, cell spreading with well-organized actin stress fibers, phosphorylation of FAK Tyr397, and neurite outgrowth. In contrast, syndecan-binding peptide AG73 (RKRLQVQLSIRT) conjugated chitosan membranes density (1.5-1500 ng/mm(2)) promoted similar biological activities at all of the concentrations tested. These results suggest that integrin-mediated cell adhesion is sensitive to the scaffold condition. To improve the function of integrin-mediated biological activities on a large amount of scaffold, we designed an A99a/AG73 mixed peptide-chitosan membrane. The mixed peptide-chitosan membrane promoted the strongest biological activities at 150-1500 ng/mm(2) of chitosan membrane. We conclude that the A99a/AG73 mixed peptide-chitosan membrane effectively interacts with both integrins and syndecans and is a useful multi-functional biomaterial.
authors.affiliation: Laboratory of Clinical Biochemistry, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo, Japan.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Hozumi
authors.forename: Dai
authors.initials: D
authors.lastname: Otagiri
authors.forename: Yuji
authors.initials: Y
authors.lastname: Yamada
authors.forename: Ayano
authors.initials: A
authors.lastname: Sasaki
authors.forename: Chikara
authors.initials: C
authors.lastname: Fujimori
authors.forename: Yuki
authors.initials: Y
authors.lastname: Wakai
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Uchida
authors.forename: Fumihiko
authors.initials: F
authors.lastname: Katagiri
authors.forename: Yamato
authors.initials: Y
authors.lastname: Kikkawa
authors.forename: Motoyoshi
authors.initials: M
authors.lastname: Nomizu
mesh_terms: D000595:Amino Acid Sequence; D002448:Cell Adhesion; D002478:Cells, Cultured; D048271:Chitosan / Q000378:metabolism*; D006801:Humans; D007797:Laminin / Q000378:metabolism*; D008567:Membranes, Artificial; D008969:Molecular Sequence Data; D010455:Peptides / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000502:physiology*
chemical_list: D007797:Laminin; D008567:Membranes, Artificial; D010455:Peptides; D011956:Receptors, Cell Surface; D048271:Chitosan