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In this study, we prepared porous nano-hydroxyapatite/polyamide 66 (n-HA/PA66) composite scaffolds utilizing injection molding method. The morphology and macrostructure as well as mechanical strength of the scaffolds were characterized. Osteoblasts (OBs) were seeded on nHA/PA66 scaffolds as experiment group, and the same amount of OBs were seeded on n-HA/PA66 plate as control group. The OB/scaffold constructs were cultured for up to 18 days, then the adhesion, proliferation and osteogenic activity of OBs were observed by scanning electron microscope (SEM) and detected by alkaline phosphatase (ALP) activity, calcium content and osteocalcin (OC) concentration. The results show that the porous n-HA/PA66 composite scaffolds are biocompatible and have no negative effects on the OBs in vitro. The scaffolds fulfill the basic requirements of bone tissue engineering scaffold, and have the potential application in orthopedic, reconstructive and maxillofacial surgery.