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Originally published as JHC exPRESS on July 11, 2007.
doi:10.1369/jhc.6A7162.2007
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Journal of Histochemistry and Cytochemistry
Volume 55 (11): 1095-1104, 2007
Copyright ©The Histochemical Society, Inc.

Immunolocalization of Sibling and RUNX2 Proteins During Vertical Distraction Osteogenesis in the Human Mandible

Lisa R. Amir, Andreas Jovanovic, Frits B.T. Perdijk, Satoru Toyosawa, Vincent Everts and Antonius L.J.J. Bronckers

Department of Oral Cell Biology, Academic Centre for Dentistry Amsterdam (ACTA), Universiteit van Amsterdam and Vrije Universiteit, Amsterdam, The Netherlands (LRA,VE,ALJJB); Department of Oral & Maxillofacial Surgery, Alkmaar Medical Center, The Netherlands (AJ); Department of Oral & Maxillofacial Surgery, Gelderse Vallei Hospital, Ede, The Netherlands (FBTP); and Department of Oral Pathology, Osaka University Graduate School of Dentistry, Osaka, Japan (ST)

Correspondence to: A.L.J.J. Bronckers, Department of Oral Cell Biology, ACTA, Vrije Universiteit, Van der Boechorststr 7, 1081 BT, Amsterdam, The Netherlands. E-mail: a.bronckers{at}vumc.nl

We tested the hypothesis that mechanical loading of human bone increases expression of the transcription factor RUNX2 and bone matrix proteins osteopontin (OPN), bone sialoprotein (BSP), dentin matrix protein-1 (DMP1), and matrix extracellular phosphoglycoprotein (MEPE). We examined this in tissue sections of atrophic mandibular bone taken from edentulous patients who had undergone distraction osteogenesis. In undistracted bone, weak to moderate staining for OPN and BSP was found in osteoblasts and bone matrix of immature woven bone. RUNX2 was also detectable in osteoblasts and in cells of the periosteum. In woven bone, but not in lamellar bone, a small number of osteocytes stained for all proteins tested. After distraction, staining intensity had increased in the existing old bone and staining was seen in more bone cells than before distraction. We also found a high expression of DMP1 and MEPE in many osteocytes embedded in woven bone and in some osteocytes of lamellar bone not seen before distraction. New bone trabeculae were forming in the fibrous tissue of the distraction gap containing all stages of intramembranous bone formation. Moderate to strong staining was seen for all five proteins tested in osteocytes located in woven bone of these trabeculae and for RUNX2, OPN, and BSP in osteoblasts lining the trabecular surfaces. We conclude that loading of atrophic human jawbone by distraction activates matrix synthesis of bone cells in and around existing bone. Increased staining of DMP1 and MEPE in osteocytes after loading is in line with the concept that these proteins may be involved in signaling the effector cells to adapt the bone structure to its mechanical demands. (J Histochem Cytochem 55:1095–1104, 2007)

Key Words: distraction osteogenesis • alveolar mandible • osteopontin • bone sialoprotein • dentin matrix protein • matrix extracellular phosphoglycoprotein • RUNX2/cbfa1 • immunohistochemistry


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