Regenerative biomedicine has emerged as a forefront area in medical research, heralding a new era of therapeutic strategies. This review delineates the integration of 2D materials (2DMs) within the area of biomedical engineering, leveraging their superior physicochemical attributes for enhance biomedical outcomes. The synergistic interaction between biomacromolecules and 2DMs is explored, demonstrating their potential to mitigate the limitations inherent to each while simultaneously augmenting their beneficial properties. In particular, incorporating 2DMs into hydrogels highlights their capability to enhance the mechanical strength of hydrogels, providing a biomimetic scaffold for tissue engineering regeneration and cancer diagnosis and therapy. An overview of the synthetic methodologies are provided for 2DMs, elucidating their interaction dynamics with biomacromolecules. The review primarily concentrates on the applications of biomacromolecule‐reinforced 2DM hydrogels across various biomedical fields, including bone tissue engineering, wound healing, neural tissue engineering, cardiac tissue engineering, as well as in the delivery of drugs and genes, cancer therapy, and biosensing technologies. Finally, the review discusses the existing challenges and future outlook for developing and using biomacromolecule‐reinforced 2DM hydrogels, underlining their transformative potential in regenerative medicine.