Copper nanoparticles were synthesized via precipitation technique using the pseudonatural cationic chitosan biopolymer as a stabilizing agent. The nanoparticles developed were successfully incorporated into the 1:1 ratio of blended chitosan: pluronic F127 polymer solution and made their nanocomposite hydrogels by solution casting method. The formed copper-based nanocomposite hydrogels were characterized by using Fourier transform infrared spectroscopy, thermogravimetric analysis, X-ray diffraction, scanning electron microscopy-energy dispersive spectroscopy and transmission electron microscopy studies. The antimicrobial activity of the fabricated nanocomposite hydrogels was tested via an inhibition zone process against both E. coli (gram-negative) and S. aureus (gram-positive) bacteria. The results conveyed that the copper-embedded chitosan-pluronic\ F127 nanocomposite hydrogels can be used effectively for antimicrobial applications as well as for wound care applications.