Chronic diabetic wounds, characterized by persistent infection and impaired tissue regeneration, remain a formidable clinical challenge. In this study, a 3D-bioprinted asymmetric bilayer scaffold was developed by integrating electrospinning and 3D bioprinting technologies to achieve "anti-infection and pro-regeneration" functionalities. The scaffold features a distinctive asymmetric architecture comprising a superficial layer (Layer S) and a basal layer (Layer B). Layer S, consisting of electrospun Cu₂O-PCL nanofibers, serves as an effective antibacterial barrier specifically targeting Methicillin-resistant Staphylococcus aureus (MRSA), while Layer B employs a 3D-bioprinted decellularized extracellular matrix (dSEM) hydrogel loaded with human bone marrow mesenchymal stem cell-derived extracellular vesicles (hBMSC-EVs) to facilitate tissue repair. Experimental results demonstrated that hBMSC-EVs significantly augmented fibroblast proliferation and migration, and the Cu₂O -doped layer exhibited potent bactericidal activity against MRSA. In db/db diabetic mice, this asymmetric composite scaffold significantly accelerated wound closure compared to standalone treatments. Histological analysis further confirmed enhanced neovascularization and accelerated extracellular matrix (ECM) reconstruction. Overall, this synergistic 3D-bioprinted bilayer system provides a high-performance strategy for the targeted management of MRSA-infected chronic diabetic wounds.