Oral and maxillofacial tumors, trauma and infections are the main causes of jaw defects, whose clinical treatment is very complicated. With the development of biological tissue engineering, many biological materials have been widely used in various fields of stomatology, and they play a very important role in the repair and replacement of maxillofacial bone defects. In this study, we intended to prepare a graphene oxide/hyaluronic acid/chitosan (GO/HA/CS) composite hydrogel with different mass ratios of GO: 0.1% (0.1% GO/HA/CS), 0.25% (0.25% GO/HA/CS), 0.5% (0.5% GO/HA/CS), and 1% (1% GO/HA/CS), prepare it into a multilayered and stable composite scaffold through 3D-printing technology, observe the surface morphology of the composite scaffold through scanning electron microscopy (SEM), and then test its physical and chemical properties, mechanical properties, water swelling rate, in vitro degradation and other material properties. Moreover, the biological performance of the GO/HA/CS composite scaffold was studied through experiments, such as cell morphology observation, cell adhesion, cell proliferation, and live-dead cell staining. The results showed that through chemical cross-linking and 3D-printing technology, a porous (pore size: 450–580 μm) and multilayered GO/HA/CS biological scaffold could be successfully constructed, and its surface was an interconnected microporous structure, and the porosity decreased (94%−40%) gradually with the increase of GO. Meanwhile, with the change in GO concentration, some mechanical properties of the scaffold could be improved, such as water swelling rate, degradation rate, and elastic modulus. In addition, the composite scaffold with the appropriate amount of GO had almost no cytotoxicity and could promote cell growth and proliferation, especially 0.25% GO/HA/CS composite scaffold. Consequently, the 0.25% GO/HA/CS composite scaffold had excellent biological material properties and good biocompatibility with osteoblasts, which may provide a new idea for the repair of jaw defects.