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With the high demand for developing novel composites with integrated performance, graphene-based nanostructures have been drawing great attention in environmental and biomedical applications because of their extraordinary physicochemical properties and biocompatibility. Although graphene oxide (GO) nanosheets exhibit some antibacterial activities, novel GO based nanostructures with enhanced antibacterial activities are highly desired. To realize this aim, polyethyleneimine (PEI) modified GO as a tetracycline hydrochloride (TCH) carrier and release platform was constructed (pGO-TCH). The nanostructures were fully characterized by TEM, AFM, FTIR and Raman spectra, which demonstrated that TCH were uniformly and compactly deposited on PEI modified GO nanosheets. The antibacterial performances of the prepared nanostructures were investigated by disk diffusion method and bacterial growth kinetics method towards Gram-positive S. aureus and Gram-negative E. coli. Results show that pGO-TCH nanostructures exhibit good antibacterial behavior. The mechanism of antibacterial activity was studied. Moreover, the nanostructures showed good cytocompatibility. This study not only highlights a promising pGO-TCH nanostructure as a candidate of graphene-based antibacterial agent, but also provides us antibacterial mechanism between bacteria and graphene-based nanomaterials. [ABSTRACT FROM AUTHOR]