2D nanomaterials show great potential in biomedical applications due to their unique physical and chemical surface properties. This review includes typical 2D nanomaterials used in bone tissue engineering (BTE), such as graphene oxide, hexagonal boron nitride, molybdenum disulfide, black phosphorus, and MXenes. Moreover, the construction methods of BTE materials with 2D nanosheets are analyzed. Before designing a BTE material, it is essential to understand the relationship between the material structure and properties. Notably, 2D nanomaterials can be hybridized with biomaterials, such as polypeptides, proteins, and polysaccharides, to improve biocompatibility and host responses. The effects of the surface properties and size of 2D nanomaterials on cellular behavior, gene expression, antibacterial properties, and cytotoxicity in BTE applications are also discussed. This work provides new design ideas and directions for constructing 2D nanomaterial-based BTE scaffolds.