Abstract: Objective：Construction of erythropoietin (EPO) contained nanofibers by tissue engineering strategies and subsequent study of its pro-osteogenic effects. Methods：Biomimetic gelatin/hydroxyapatite (Gel/HA) nanofibrous scaffold was fabricated by electrostatic spinning method. Erythropoietin (EPO) loaded Gel/HA scaffold was prepared by freeze-dried method after crosslinking of Gel/HA scaffold. The release profile of EPO was detected by ELISA. The osteogenic effects of Gel/HA/EPO were detected by q-PCR. A 5mm diameter calvarial bone defect model was created in rat and scaffolds were implanted. Cone-Beam CT and histological technique were used to evaluate the bone repair effects. Results：SEM results showed that all the scaffolds were reticular and intertwined structure. There was some deposition of EPO protein on the surface of Gel/HA/EPO scaffold. The release profile of EPO showed an obvious burst release phenomenon, and only a small amount of EPO remnants kept releasing slowly. q-PCR results showed that EPO can promote osteoblast differentiation by promoting the expression of Col1 and Osterix. In vivo study showed that the Gel/HA/EPO group had a large amount of new bone formation both in 6-week and 12-week post implantation, which obviously was higher than those of the blank control group and Gel/HA group; Besides, the new bone formation in the Gel/HA group was also significantly higher than that of the blank control group. Conclusions：EPO loaded biomimetic Gelatin/HA nanofibers is an excellent scaffold material for bone tissue engineering.