In this study, bamboo fiber was first designed to incorporate into nano-hydroxyapatite/poly(lactic-co-glycolic) to obtain a new composite scaffold of bamboo fiber/nano-hydroxyapatite/poly(lactic-co- glycolic) (BF/n-HA/PLGA) by freeze-drying method. The effect of their components and some factors consisting of different freeze temperatures, concentrations, and pore-forming agents on the porous morphology, porosity, and compressive properties of the scaffold were investigated by scanning electron microscope, modified liquid displacement method, and electromechanical universal testing machine. The results indicated that the 5% BF/30% n-HA/PLGA composite scaffold, prepared with 5% (w/v) high concentration and frozen at -20 °C without pore-forming agent, had the best ideal porous structure and porosity as well as compressive properties, which far exceed those of n-HA/PLGA composite scaffold. In addition, the in vitro simulated body fluids soaking and cell culture experiment showed the addition of BF into the scaffold accelerated the BF/n-HA/PLGA composite scaffolds degradation and exhibited good cytocompatibility, including attachment and proliferation. All the results of the study show that BF has improved the properties of n-HA/PLGA composite scaffolds and BF/n-HA/PLGA might have a great potential for bone tissue engineering scaffold.