The present work reports low cost, green synthesis of Nanocellulose (NC) nanoparticles. The biosynthesized nanoparticles were characterized by transmission electron microscopy (TEM), X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (ATR-FTIR). The synthesized NC nanoparticles were pure, predominantly spherical in shape with size ranging from 25 nm. The biosynthesized NC nanoparticles have been used for antibacterial and in vitro applications. The antibacterial activity of the prepared CS/PVA/MC-NC1,CS/PVA/MC-NC2, and CS/PVA/MC-NC3 nanocomposites samples was tested against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) due to disk diffusion method. When adding the NC nanoparticles both bacteria were very good inhibition zone was obtained. The transmission NC micrographs depicts spherical with uniform shape and good crystallinity and are composed of nanoparticles with a diameter less than 25 nm the average cube size was 100 nm. The AFM thickness of the CS/PVA/MC-NC1 scaffold was, estimated from the AFM image, was about 10-20 nm and a roughness-like structure was observed. The FESEM film exhibits a scaffold exhibited porous structures. The excellent cell viability of the composite scaffolds was attributed to the good biocompatibility of the CS/PVA/MC-NC3 as well as green fabrication process of the scaffolds. MTT analysis exposed that the samples did not have any toxicity. Since these positive points, these two kinds of scaffolds show appropriate properties for attachment, proliferation, and tendency to form group from L929 cells. In this work, we have prepared nanocellulose by high pressure homogenization process and the resultant nanocellulose was evaluated as fibers in CS/PVA/MC films. (C) 2017 Elsevier B.V. All rights reserved.