Infectious bone defects pose a significant challenge in orthopedics by hindering healing and vascularization. This study explored the impact of fibroin thermosensitive hydrogel on osteogenesis, inflammatory response, and angiogenesis as a potential biomaterial for bone regeneration in osteomyelitis treatment. The biocompatibility of the hydrogel by live/dead staining revealed a high number of viable osteoblast cells after 14 days. ALP activity was significantly increased in all hydrogel formulations, with F3 showing the highest levels of total protein content and calcium deposition, indicating more effective osteogenesis. Gene expression analysis of the osteogenesis-related genes demonstrated that