Regenerative biomedicine has become a cutting-edge field in medical research, heralding the arrival of a new era of treatment strategies. This review provides an overview of the integration of two-dimensional materials (2DM) in the field of biomedical engineering, utilizing their superior physicochemical properties to enhance biomedical achievements. Explored the synergistic interactions between biomolecules and 2DM, demonstrating their potential to enhance their beneficial properties while reducing their inherent limitations. In particular, the incorporation of 2DM into hydrogels highlights their ability to enhance the mechanical strength of hydrogels, providing bionic scaffolds for tissue engineering regeneration and cancer diagnosis and treatment. This article provides an overview of the synthesis methods of 2DMs and elucidates their interaction kinetics with biomolecules. This review focuses on the application of biomacromolecule enhanced 2DM hydrogel in various biomedical fields, including bone tissue engineering, wound healing, neural tissue engineering, heart tissue engineering, drug and gene delivery, cancer treatment and biosensor technology. Finally, this review discusses the current challenges and future prospects of developing and using biomacromolecule enhanced 2DM hydrogels, and emphasizes their potential for innovation in regenerative medicine.