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The aim of the present study was to develop antibacterial cellulose (cotton) nanocomposite fabrics (CNCFs) with in situ generated silver nanoparticles using medicinal plant Vitex leaf extract. The developed CNCFs were characterized by scanning electron microscope (SEM), Fourier transform infrared (FT-IR) spectroscopy, X-ray diffraction (XRD) and antibacterial tests. Further, these CNCFs possessed good antibacterial activities. These CNCFs prepared using simple and environmentally friendly method can be considered for medical applications in, such as, surgical aprons, wound cleaning, wound dressing, and hospital bed materials.