Silk fibroin hydrogels occupy an essential position in the biomedical field due to their remarkable biological properties, excellent mechanical properties, flexible processing properties, as well as abundant sources and low cost. Herein, we introduce the unique structures and physicochemical characteristics of silk fibroin, including mechanical properties, biocompatibility, and biodegradability. Then, various preparation strategies of silk fibroin hydrogels are summarized, which can be divided into physical cross‐linking and chemical cross‐linking. Emphatically, the applications of silk fibroin hydrogel biomaterials in various biomedical fields, including tissue engineering, drug delivery, and wearable sensors, are systematically summarized. At last, the challenges and future prospects of silk fibroin hydrogels in biomedical applications are discussed.