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A poly(γ⁻glutamic acid)/β⁻tricalcium phosphate (γ⁻PGA/β⁻TCP) composite fibrous mat was fabricated using the electrospinning technique as a novel bone substitute. The mat was then cross-linked with cystamine in the presence of 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide to improve its water-resistant ability. Scanning electron micrographs revealed that the γ⁻PGA/β⁻TCP fibers had a uniform morphology with diameters ranging from 0.64 ± 0.07 µm to 1.65 ± 0.16 µm. The average diameter of the fibers increased with increasing cross-linking time. Moreover, increasing the cross-linking time and decreasing the γ⁻PGA/β⁻TCP weight ratio decreased the swelling ratio and in vitro degradation rate of the composite fibrous mat. In vitro experiments with osteoblast-like MG-63 cells demonstrated that the mat with a γ⁻PGA/β⁻TCP weight ratio of 20 and cross-linked time of 24 h had a higher alkaline phosphatase activity and better cell adhesion. Furthermore, the rat cranial bone defect was created and treated with the γ⁻PGA/β⁻TCP composite fibrous mat to evaluate its potential in bone regeneration. After 8 weeks of implantation, micro computed tomography showed that the γ⁻PGA/β⁻TCP composite fibrous mat promoted new bone growth. These observations suggest that the γ⁻PGA/β⁻TCP composite fibrous mat has a potential application in bone tissue engineering.