Recently, tissue engineering has developed into a powerful tool for repairing and reconstructing damaged tissues and organs. Tissue engineering scaffolds play a vital role in tissue engineering, as they not only provide structural support for targeted cells but also serve as templates that guide tissue regeneration and control the tissue structure. Over the past few years, owing to unique physicochemical properties and excellent biocompatibility, various types of two-dimensional (2D) nanomaterials have been developed as candidates for the construction of tissue engineering scaffolds, enabling remarkable achievements in bone repair, wound healing, neural regeneration, and cardiac tissue engineering. These efforts have significantly advanced the development of tissue engineering. In this review, we summarize the latest advancements in the application of 2D nanomaterials in tissue engineering. First, each typical 2D nanomaterial is introduced briefly, followed by a detailed description of its applications in tissue engineering. Finally, the existing challenges and prospects for the future of the application of 2D nanomaterials in tissue engineering are discussed.