The basis of lengthening bones at their anatomic locations has been defined in detail by many authors. The possibility of distraction osteogenesis in vascularized bone grafts and the role of periosteum was investigated in 35 growing New Zealand White rabbits in four experimental groups: group 1, the scintigraphic assessment group (N = 4); group 2, the distraction group with intact periosteum (N = 15); group 3, the control group without periosteum in the distraction area (N = 6); and group 4, the control group in which the flaps were wrapped in silicone leaves (N = 10). Scintigraphy was used to evaluate the survival of the bone graft. Metatarsal fasciosteal flaps were fixed by external fixators and then subjected to osteotomy. Distraction was started after 10 days of osteotomy and continued at 0.5 mm/24 hr for 10 days, with a total elongation of 4 to 6 mm. Radiological examination at day 10 of starting the distraction, and both radiological and histological examinations at days 15, 20, and 30 were performed. In group 1, positive scintigraphic images of the flaps were obtained. In group 2, progressive calcification was detected radiologically and mixed-type ossification was observed histologically. However, in group 3, distraction failed. Group 4 was excluded from the study due to silicone exposition. The authors demonstrated that vascularized bone segments can be lengthened, and provided information about the importance of periosteum in distraction osteogenesis.