The purpose of this study is to analyze osteogenic activity of human osteoblastic MG63 cells on the bone-like self-organized hydroxyapatite/collagen (HAp/Col) nanocomposite sponge cultured by a pressure/perfusion technique using collagen sponge as a control. Histological analyses, alkaline phosphatase (AlkP) protein analysis and real-time polymerase chain reaction (PCR) analyses for AlkP and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) to evaluate the osteogenic activity of MG63. The MG63 cells were attached well and showed good proliferation on the HAp/Col sponge as well as in the control. The MG63 cells on HAp/Col sponge demonstrated higher osteogenic activity than those on the control. The results suggested that the HAp/Col sponge is expected to be a good scaffold for bone tissue engineering.