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The nerve conduit with biofunctionalities can regulate neurite outgrowth, as well as the migration, proliferation, and myelination activity of Schwann cells. In the present study, polycaprolactone (PCL) conduits are coated with Naphthalene-phenylalanine-phenylalanine-glycine-arginine-glycine-aspartic (Nap-FFGRGD) and Naphthalene-phenylalanine-phenylalanine-glycine-cysteine-aspartic-proline-glycine-tyrosine-isoleucine-glycine-serine-arginine (Nap-FFGCDPGYIGSR) by self-assembly. In vitro studies demonstrate that arginine-glycine-aspartic (RGD) and tyrosine-isoleucine-glycine-serine-arginine (YIGSR) are capable of synergistically enhancing the ability of PCL to support the adhesion and proliferation of Schwann cells, as well as increasing neurite outgrowth from dorsal root ganglions explants. This synergistic effect may occur via the activation of both the phosphoinositide 3-kinase/protein kinase B and mitogen-activated protein kinase/extracellular signal-regulated protein kinase pathways. RGD/YIGSR modifications demonstrate beneficial effects across a 15 mm sciatic nerve gap in axonal regeneration and functional recovery. In addition, increased vascularization is observed in the RGD/YIGSR-PCL group, which might contribute to their beneficial effects on nerve regeneration. These findings indicate the potential of the RGD/YIGSR-PCL conduit to promote axonal regeneration and functional recovery, making the RGD/YIGSR-PCL conduit an attractive candidate for the treatment of a critical nerve defect.