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Local OPG Gene Transfer to Periodontal Tissue Inhibits Orthodontic Tooth Movement

¹ Division of Orthodontics and Dentofacial Orthopedics, and ² Division of Oral Dysfunction Science, Department of Oral Health and Development Sciences, Graduate School of Dentistry, Tohoku University, 4-1 Seiryo-machi, Aoba-ku, Sendai 980-8575, Japan;

View larger version (66K): [in this window] [in a new window]   Figure 1. Local gene transfer area and systemic effects. (a) Schema of local gene transfer. Vector-solution was injected into the sub-periosteal area, adjacent to the first molars. Bu: buccal side. Pa: palatal side. (b) Body weight curve of experimental animals. The body weights of the experimental animals were measured throughout the experiment. There were no significant differences among the 3 groups. The results are expressed as the mean ± SD. N = 3, 8, and 9, respectively (control group, tooth-movement group, OPG-transfection group). (c) X-ray photograph of proximal rat tibiae. The densitometric patterns inside the standardized white box area were measured. (d) Bone mineral density values of tibiae of both groups. The results are expressed as the mean ± SD (N = 8). There were no significant differences among the groups.

View larger version (79K): [in this window] [in a new window]   Figure 2. In vitro assay of pcDNA-mOPG. (a) RT-PCR analysis of NIH3T3 cells. RT-PCR analysis was performed with cDNA from OPG-transfected NIH3T3 cells and control NIH3T3 cells. The results shown are from 1 representative independent experiment out of 3. C, control NIH3T3 cells; T, OPG-transfected NIH3T3 cells; L, DNA ladder. (b) Western blot analysis with antibody against OPG. L, protein ladder; Control, cell lysate from control NIH3T3 cells; Transfect, cell lysate from OPG-transfected NIH3T3 cells. The results shown are from 1 representative independent experiment out of 3. (c) Resorption assay (photos). Upper panel: Osteologic® discs cultured with normal culture medium and 1,25-(OH)2D3. Lower panel: Osteologic® discs cultured with 50% collected culture medium and 1,25-(OH)2D3. Arrowhead indicates resorption pit. The areas of the resorption pits from randomly selected fields were measured. The results shown are from 1 representative experiment out of 8. Bar = 100 µm. (d) Resorption assay (resorbed area of Osteologic® discs). The areas of the resorption pits from randomly selected fields were measured. The resorbed areas of 24 fields were measured. * p < 0.05.

View larger version (107K): [in this window] [in a new window]   Figure 3. Immunohistochemical analysis of OPG. Immunofluorescence analyses of serial sections were performed with anti-OPG antibodies. Images (a) to (c) are photographs of the palatal sides of first molars. Fluorescence image and phase-contrast image were superimposed on the software (Adobe Photoshop 5.0, San Jose, CA, USA). (a) Photograph of the tooth-movement group. Representative photographs of 6 sections are shown. (b) Photograph of the OPG-transfection group. Representative photographs of 5 sections are shown. (c) Photograph of the mock group. Representative photographs of 4 sections are shown. Bar = 100 µm. Bu, buccal side. Pa, palatal side. (d) Combined photograph of the OPG-transfection group. Series of the fluorescent photographs taken under the same exposure conditions were combined. Strong OPG expression was observed at M1. (e) Photograph of the OPG-transfection group stained with hematoxylin-eosin. About the same field as shown in Fig. 3d is presented. Bar = 1 mm.

View larger version (58K): [in this window] [in a new window]   Figure 4. Osteoclastogenesis and tooth movement. Photo images of osteoclasts. TRAP-positive multinucleated cells were observed around the teeth in the tooth-movement group (a) and in the OPG-transfection group (b). Periodontal tissues of the mesiopalatal roots of the upper first molars were examined in serial cross-sections of the molars at a bifurcation level. TRAP-positive multinucleated cells that formed resorption lacunae on the alveolar bone surface adjacent to the mesiopalatal roots of the upper first molars were counted. Representative photographs of 6 sections of both groups are shown. Bar = 100 µm. (c) Number of osteoclasts in the periodontium. TRAP-positive multinucleated cells that formed resorption lacunae on the alveolar bone surface adjacent to the mesiopalatal roots of the upper first molars were counted. The results are expressed as the mean ± SD (N = 6). (d) Time course of changes in tooth movement. N = 8 and 9, respectively (tooth-movement group, OPG-transfection group). No tooth movement to the palatal side was observed in both control and mock groups (data not shown). (e) Data are presented as percent inhibition of tooth movement (OPG-transfection group/tooth-movement group) in each day. *P < 0.05; **P < 0.01; ***P < 0.001.

Journal of Dental Research, Vol. 83, No. 12, 920-925 (2004)
DOI: 10.1177/154405910408301206


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