Supramolecular assembly loading drug as biomedical materials is a research hotspot. Herein, we reported a supramolecular electrospun assembly constructed via the hydrophobic and hydrogen bonding interaction. The obtained results showed that the assembly by supramolecular electrospinning not only increased the interactions of multiple antibacterial active species including antibiotics, cationic polymers, and silver to form a flexible membrane with good mechanical strength but also indicated the dual effects of rapid doxycycline and polyethyleneimine release as well as a sustained Ag release. Interestingly, the assembly showed not only good degradability but also a high bacteriostatic efficacy toward Escherichia coli (E. coli) up to 99.9%. More importantly, the in vivo wound healing assay indicated that the assembly could promote the healing of uninfected, E. coli-infected, and even methicillin-resistant staphylococcus aureus-infected wounds. The current research provides a novel approach to construct a supramolecular assembly by electrospinning mechanically induced strong noncovalent interaction.