Hydrogels catalyzed by horseradish peroxidase (HRP) serve as an efficient and effective platform for biomedical applications due to their mild reaction conditions for cells, fast and adjustable gelation rate in physiological conditions, and an abundance of substrates as water-soluble biocompatible polymers. In this review, we highlight the tunable characteristics and use of the HRP-catalyzed hydrogels and provide a brief overview of various substrates employed in the HRP system for different biomedical applications of the resultant hydrogels. In addition, we discuss and summarize the biocompatibility, possible functionalization, and biofabrication process. Finally, the future prospective of the HRP crosslinking system is highlighted with biomedical applications.