This study presents a hybrid 3D printed scaffold with QK and KP peptides to enhance bone regeneration in cranial defects. The scaffold consists of vertically aligned cryogel fibers (ACFs) that guide cell penetration into the defect area, followed by the regulation of cell function by the peptides to promote vascularization and osteogenic differentiation. The dual peptide-modified scaffold significantly increased regenerated bone volume and surface coverage compared to the collagen sponge control. In vitro and in vivo studies demonstrated that the scaffold enhanced cell recruitment, angiogenesis, and osteogenic differentiation. The KP peptide promoted osteogenic differentiation, while the QK peptide enhanced angiogenesis. The dual peptide-modified scaffold showed sustained bone regeneration and reduced bone resorption, making it a promising strategy for bone regeneration.This study presents a hybrid 3D printed scaffold with QK and KP peptides to enhance bone regeneration in cranial defects. The scaffold consists of vertically aligned cryogel fibers (ACFs) that guide cell penetration into the defect area, followed by the regulation of cell function by the peptides to promote vascularization and osteogenic differentiation. The dual peptide-modified scaffold significantly increased regenerated bone volume and surface coverage compared to the collagen sponge control. In vitro and in vivo studies demonstrated that the scaffold enhanced cell recruitment, angiogenesis, and osteogenic differentiation. The KP peptide promoted osteogenic differentiation, while the QK peptide enhanced angiogenesis. The dual peptide-modified scaffold showed sustained bone regeneration and reduced bone resorption, making it a promising strategy for bone regeneration.