Treatment of bone defects in osteoporotic patients with bone substitutes is difficult, due to insufficient osseointegration. The development of appropriate biomaterials to solve the problem requires the assessment of the material performance in an osteoporotic environment, which is rarely investigated. Herein, nanostructured biphasic calcium phosphate (nBCP) ceramics were prepared via the incorporation of hydroxyapatite nanoparticles (HANPs) into porous biphasic CaP (BCP) substrates, leading to an increase of over 500% in the specific surface area. Primary osteoblasts harvested from osteoporotic rats were cultured on the nBCP ceramics, and it was found that the osteoblast functions, including proliferation, alkaline phosphatase activity, osteocalcin secretion and expression of osteogenic genes, were significantly enhanced compared with osteoblasts grown on non-nanostructured BCP ceramics. To further assess the osteoinduction ability, the ceramics were implanted in the femur of osteoporotic rats. Compared to the rats implanted with non-nanostructured BCP ceramics, a higher amount of mechanically matured bone was newly formed in the rats with nBCP ceramics after 6 weeks of implantation. Such enhanced osteoinduction ability of the nBCP ceramics may be due to the incorporated HANPs, as well as the nanostructured topography induced by the HANPs. These results indicate good in vitro and in vivo osteoinductivity of the nBCP ceramics in an osteoporotic environment and offer potential benefits for treating bone defects in osteoporotic patients.