Distraction osteogenesis has become a widely used clinical approach in the treatment of craniofacial and orthopedic disorders. The exact biological mechanism of bone formation remains illusive, however. The aim of this study was to evaluate the role of bone morphogenetic protein-2, bone morphogenetic protein-4, and transforming growth factor-beta superfamily-related postreceptor signaling glycoproteins Smads 1 through 5 in distraction osteogenesis. Twelve sheep randomly divided into two groups were distracted to 24 mm at 1 or 4 mm/d using a submandibular osteotomy and an external distractor. After a 5-week fixation period, the mandibles were harvested. Employing immunohistochemical techniques, the sections were investigated for the previous antigens. Osteoblasts and periosteal lining cells were strongly positive. The matrix did not stain for the antigens investigated. Osteocytes demonstrated weak staining for the antigens. No significant difference between the groups was noted. In fracture healing, bone morphogenetic proteins 2 and 4 have been localized to the cambial layer of the periosteum, where healing occurs by intramembranous ossification. Their diffuse staining of the osteoblasts in the distracted region supports a similar role in distraction osteogenesis, where bone formation is predominantly through intramembranous ossification. Furthermore, bone morphogenetic proteins 2 and 4 have been demonstrated to promote mesenchymal cell conversion to osteoblasts. This is similar to the process observed in distraction osteogenesis. The presence of related Smads confirms postreceptor activity of these bone morphogenetic proteins in the process of distraction osteogenesis. This study supports induction of bone morphogenetic proteins 2 and 4, their related postreceptor signaling system (Smads), and intramembranous bone formation associated with mechanical strain in distraction osteogenesis.