In this work, composite hydrogel consisting of poly(vinyl alcohol), graphene and silver nanoparticles, (Ag/PVA/Gr), was prepared by the immobilization of silver nanoparticles (AgNPs) in PVA/Gr hydrogel matrix in two steps. The first step was cross linking of the PVA/Gr colloid solution by the freezing/thawing method, while in the second step, in situ electrochemical method was used to incorporate AgNPs inside the PVA/Gr hydrogel matrix. We used UV-vis spectroscopy, cyclic voltammetry, Raman spectroscopy, DSC analysis, as well as test of cytotoxicity and antibacterial activity, to characterize obtained Ag/PVA/Gr hydrogel. It was shown that graphene-based composite hydrogel with incorporated AgNPs is non toxic biomaterial with antibacterial activity, with a potential for use in biomedical purposes.