Abstract Repair of critical bone defects is a challenge in the orthopedic clinic. 3D printing is an advanced personalized manufacturing technology that can accurately shape internal structures and external contours. In this study, the composite scaffolds of polylactic acid (PLA) and nano-hydroxyapatite (n-HA) were manufactured by the fused deposition modeling (FDM) technique. Equal mass PLA and n-HA were uniformly mixed to simulate the organic and inorganic phases of natural bone. The suitability of the composite scaffolds was evaluated by material characterization, mechanical property, and in vitro biocompatibility, and the osteogenesis induction in vitro was further tested. Finally, the printed scaffold was implanted into the rabbit femoral defect model to evaluate the osteogenic ability in vivo. The results showed that the composite scaffold had sufficient mechanical strength, appropriate pore size, and biocompatibility. Most importantly, the osteogenic induction performance of the composite scaffold was significantly better than that of the pure PLA scaffold. In conclusion, the PLA/n-HA scaffold is a promising composite biomaterial for bone defect repair and has excellent clinical transformation potential.