The aggregation of multi-walled carbon nanotubes (MWCNT) into the polymer matrix and weak bactericidal property of MWCNT usually limit the application of MWCNT in the infection resistance. To solve the above problem, vancomycin hydrochloride modified multi-walled carbon nanotube (Van-MWCNT) via the chemical reaction of the MWCNT's inherent carboxyl group and the Van's amide group was well designed and synthesized. Then, Van-MWCNT was in-situ anchored onto thermoplastic polyurethane (TPU) electrospun nanofibers (TPU/Van-MWCNT) by ultrasonication assistance to achieve the bactericidal property. The above preparation process was facile, non-toxic with the green solvent-water as medium. At the same time, it also effectively reduced the aggregation of Van-MWCNT into the electrospun nanofibers. The minimal inhibitory concentration (MIC) of Van-MWCNT against S. aureus was obviously lower than that of MWCNT. And the TPU/Van-MWCNT exhibited excellent antibacterial properties evaluated through the spread plate test and field emission scanning electron microscope (FESEM). The as-prepared antibacterial TPU electrospun nanofibers have potential application in wound dressing. (C) 2017 Elsevier Ltd. All rights reserved.