Microsurgical vascularized bone transfer has the disadvantages of limitation of available donor sites, loss of donor function, and the possibility of donor site defects or deformity. To overcome these shortage of current microsurgical tissue transfer, the method of creating the neovascularized free flap has been introduced. Potentially, this technique must be an innovation in providing the free vascularized bone grafts that are not limited by natural vascular anatomy. But, as could be imagined technique resulted in unavoidable donor bone defect and additional operation for harvesting the autologous bone. The purpose of this study was to evaluate the efficacy of demineralized allogeneic bone as a possible substitute for autologous bone in fabricating the neo-osseous flap. By histologic, microangiographic and radioisotope method, the viability and vascularity of neo-osseous flap, which has been fabricated using allogeneic bone or autologous bone, was assessed in rat model. After 6 weeks, demineralized allogeneic bone showed consistent bone formation and neovascularization. The clinical and microscopic findings of demineralized allogeneic bone group were inferior to those of autogenous bone with regard to bone regeneration. The amount of bone blood floow per dry weight of demineralized allogeneic bone group was significantly higher than that of autogenous bone, even higher that of control intact iliac bone. In conclusion, findings supported that allogeneic bone could be the potential substitute for autologous bone source in creating a prefabricated neo-osseous flap.