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Runx2 Regulates Endochondral Ossification in Condyle during Mandibular Advancement

¹ Hard tissue biology and repair research group and Orthodontics, Faculty of Dentistry, The University of Hong Kong, Prince Philip Dental Hospital, 34 Hospital Road, Hong Kong SAR, China; and
² Department of Orthodontics, School of Stomatology, Shanghai Second Medical University, Shanghai, China;

View larger version (137K): [in a new window]   Figure 1. Alcian blue and PAS staining showed an overview of the rat’s temporomandibular joint during natural growth (A: 42 days old) and during mandibular advancement (B: 7 days after the experiment; note the forward positioning of the condyle). Two measurement frames that overlapped at the erosive front in the posterior condyle were used for quantitative analysis (B). In situ hybridization (ISH, C and D, arrows) and immunohistochemical staining (IHC, F and G, arrowheads) showed Runx2 expression in the rat’s condylar cartilage during natural growth (C and F: 49 days old) and mandibular advancement (D and G: 14 days after the experiment). In situ hybridization with sense probe (E) and immunostaining omitting primary antibody (H) served as negative control. Scale bars: 100 µm.

View larger version (177K): [in a new window]   Figure 2. High magnification showed the expression of Runx2 (A and D, in situ hybridization, arrows; B and E, immunohistochemical staining, arrowheads) and type X collagen (C and F: immunohistochemical staining) in the rat’s condylar cartilage during natural growth (A,B,C: 56 days old) and during mandibular advancement (D,E,F: 21 days after the experiment).

View larger version (22K): [in a new window]   Figure 3. The temporal pattern of the expression of Runx2 (A, n = 10), type X collagen (B, n = 10), BrdU-labeled cells in the erosive zone (C, n = 5), and the amount of new bone formation (D, n = 10) in the mandibular condyle during natural growth (Cont.) and during mandibular advancement (Exp.). Values were mean ± SD. Significant difference between control and experimental animals was marked with asterisks (*p < 0.05, **p < 0.01, ***p < 0.001).

View larger version (174K): [in a new window]   Figure 4. BrdU labeling in 56-day-old rats showed the differentiation of osteoblasts. When BrdU was administered 1 hr before death (A), the labeled cells were accumulated in the center of the blood vessels. The disc-shaped osteoblasts lining the subchondral bone surface were not labeled (A, arrowheads). When BrdU was administered 3 days before death (B), almost all the labeled cells in the marrow cavity were skeletal progenitor cells migrating toward the cavity wall (B, arrows). The differentiated osteoblasts on the bone surface were labeled (B, arrowheads). The labeled cells in the erosive front were multinuclear chondroclasts (C) with strong TRAP activities (D). Three-day BrdU labeling showed more osteoblasts and chondroclasts in the erosive zone during mandibular advancement (F: 14 days after the experiment) than during natural growth (E: 49 days old).