In vitro studies have demonstrated nanofibrous scaffolds have promising clinical applications in the management of bone defects. However, few have assessed bone regeneration based on macroscopic observation. This study was aimed to assess bone regeneration by macroscopic osteogenesis and observation of bone density by X-ray imaging. Using a co-electrospinning method, we fabricated bilayer fibrous scaffolds consisting of an outer layer of polycaprolactone/polylactic acid to provide mechanical strength and an inner layer of collagen I/polylactic acid to promote bone induction. The scaffolds were characterized by high-magnification scanning electron microscopy and tensile strength tests. Bone regeneration of the scaffold was evaluated in vivo by macroscopic osteogenesis and observation of bone density by X-ray imaging. The results illustrated that the bilayer scaffolds were porous, similar to natural collagen found in the extracellular matrix. Additionally, the scaffolds showed sufficient mechanical strength for surgical procedures. The bilayer scaffolds also showed higher scores of macroscopic osteogenesis and bone density evaluation on X-ray imaging than controls after implantation in vivo. This study illustrated the fabricated bilayer fibrous scaffolds may have promising practical applications for the management of bone defects in clinical practice and could be measured by clinical methods.