Cell surface receptor-specific scaffold requirements for adhesion to laminin-derived peptide–chitosan membranes

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 αvβ3 integrin-binding peptide A99a (ALRGDN) was conjugated on chitosan membranes of varying density (1.5–1500 ng/mm2), cell adhesion was altered depending on the amount of chitosan. 3 or 30 ng/mm2 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/mm2) 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/mm2 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.