Enhancement of physical and mechanical properties of the scaffolds is achieved via various methods including cross-linking and incorporation of nano particles. In the present research chitosan-based scaffolds firstly were reinforced with the incorporation of the graphene oxide (GO) nanoparticles. The GO nanoparticles were synthesized from graphite successfully and were identified by TGA, XRD, SEM and FTIR analytical methods. Nanocomposite scaffolds based on chitosan with different percentages of the GO were prepared. The chitosan-GO nanocomposite scaffolds were then simultaneously sterilized and cross-linked in an autoclave and comprehensively characterized. In XRD measurements, the absence of the peak at 10° related to GO, is evidence that the GO layers are exfoliated. With increasing the GO concentration from 0 to 0.1, 0.2 and 0.3%, physical and mechanical properties were improved, considerably. Seeding of the human articular chondrocytes on the nanocomposite scaffolds showed an increased proliferation with augmentation of the GO percentage particularly in prolonged cultivation periods (14 days). Investigation of the human articular chondrocyte morphology revealed a more spherical morphology of the cells on the cross-linked scaffolds for 21 days of culture in vitro.