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Immunohistochemical Localization of Bone Morphogenetic Protein–signaling Smads during Long-bone Distraction Osteogenesis

Tasima Haque, Manuela Mandu-Hrit, Frank Rauch, Dominique Lauzier, Maryam Tabrizian and Reggie C. Hamdy

Shriners Hospital for Children and Division of Orthopaedics, Montreal Children's Hospital (MM-H,FR,DL,RCH), Faculty of Dentistry (MM-H,MT), Department of Biomedical Engineering (MT), and Centre for Biorecognition and Biosensors (MT), McGill University, Montréal, Québec, Canada

Correspondence to: Reggie C. Hamdy, MD, Shriners Hospital for Children, 1529 Cedar Avenue, Montréal, Québec, Canada H3G 1A6. E-mail: rhamdy{at}shriners.mcgill.ca

In this study we investigated the expression of bone morphogenetic protein (BMP)-signaling Smads in distraction osteogenesis (DO). Osteotomy of the right tibia was performed in 14 skeletally mature white New Zealand male rabbits. Lengthening was started 1 week later at a rate of 0.5 mm/12 hr and was maintained for 3 weeks. Expression of Smad proteins 1, 4, 5, 6, 7, and 8 and Smad ubiquitin regulatory factors (Smurfs) 1 and 2 was evaluated in the distracted zone using immunohistochemistry. Expression of receptor-regulated Smads (R-Smads) 1, 5, and 8 showed a significant increase during the distraction phase, followed by a gradual decrease during the consolidation phase. Smad 4 showed significant expression during both distraction and the beginning of the consolidation phase. Smad 6 and Smad 7 were highly expressed during the consolidation phase. Staining for both Smurfs 1 and 2 was maximal at the end of the distraction period. Staining for all proteins was most intense in chondrocyte and fibroblast-like cells. Expression pattern of R-Smads correlated with our previously reported expression pattern of BMPs 2, 4, and 7 and their receptors. These results therefore suggest a role for the whole BMP signaling pathway including the Smad proteins in DO. (J Histochem Cytochem 54:407–415, 2006)

Key Words: bone morphogenetic proteins • distraction osteogenesis • immunohistochemistry • Smad

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