Disintegrin-Derived Cyclic RGD Surfaces Enable Integrin α5β1-Mediated Rapid Cell Adhesion

Abstract Integrin-mediated cell adhesion is a key determinant of biomaterial performance in tissue engineering and regenerative medicine. Although Arg–Gly–Asp (RGD) peptides are widely used as minimal cell-adhesive ligands, conventional RGD-functionalized surfaces predominantly engage αv-class integrins, limiting the ability to exploit the distinct functions of other integrin subtypes in cell–material interactions. Despite advances in ligand design, it remains unclear how RGD-flanking sequences in natural ligands can be leveraged to achieve subtype-selective integrin engagement. In this study, RGD-flanking sequences derived from snake venom disintegrins were systematically analyzed to identify motifs suitable for integrin-selective peptide design. Based on sequence features of the RGD loop region, thioether-cyclized peptides incorporating representative motifs were designed and synthesized, and immobilized on functionalized surfaces. Among the peptides evaluated, the cARGDNP-functionalized surface exhibited a distinct integrin-binding profile, engaging integrin α5β1 in addition to αv integrins. Time-resolved adhesion assays demonstrated that the cARGDNP surface significantly accelerated the initial phase of cell attachment, and this effect was attenuated by α5β1 blockade. These findings indicate that cARGDNP, derived from disintegrin-inspired RGD-flanking sequences, functions as a cyclic peptide ligand that enables integrin α5β1 engagement and promotes rapid initial cell adhesion on engineered surfaces.