Tadahiko Utsunomiya

Low-energy laser stimulates tooth movement velocity via expression of RANK and RANKL

OBJECTIVE Recent studies have demonstrated that low-energy laser irradiation stimulates bone formation in vitro and in vivo. However, very little is known about the effects of laser irradiation on osteoclastogenesis. The receptor activator of the nuclear factor-kB (RANK) / RANK ligand (RANKL) / osteoprotegerin (OPG) system is essential and sufficient for osteoclastogenesis. The present study was designed to examine the effects of low-energy laser irradiation on expressions of RANK, RANKL, and OPG during experimental tooth movement. DESIGN To induce experimental tooth movement in rats, 10 g of orthodontic force was applied to the molars. Next, a Ga-Al-As diode laser was used to irradiate the area around the moved tooth and the amount of tooth movement was measured for 7 days. Immunohistochemical staining with RANK, RANKL, and OPG was performed. Real time PCR was also performed to elucidate the expression of RANK in irradiated rat osteoclast precursor cells in vitro. RESULTS In the irradiation group, the amount of tooth movement was significantly greater than in the non-irradiation group by the end of the experimental period. Cells that showed positive immunoreactions to the primary antibodies of RANKL and RANK were significantly increased in the irradiation group on day 2 and 3, compared with the non-irradiation group. In contrast, the expression of OPG was not changed. Further, RANK expression in osteoclast precursor cells was detected at an early stage (day 2 and 3) in the irradiation group. CONCLUSION These findings suggest that low-energy laser irradiation stimulates the velocity of tooth movement via induction of RANK and RANKL.

Low-energy laser irradiation facilitates the velocity of tooth movement and the expressions of matrix metalloproteinase-9, cathepsin K, and alpha(v) beta(3) integrin in rats

It has previously been reported that low-energy laser irradiation stimulated the velocity of tooth movement via the receptor activator of nuclear factor kappa B (RANK)/RANK ligand and the macrophage colony-stimulating factor/its receptor (c-Fms) systems. Matrix metalloproteinase (MMP)-9, cathepsin K, and alpha(v) beta(3) [alpha(v)beta3] integrin are essential for osteoclastogenesis; therefore, the present study was designed to examine the effects of low-energy laser irradiation on the expression of MMP-9, cathepsin K, and alpha(v)beta3 integrin during experimental tooth movement. Fifty male, 6-week-old Wistar strain rats were used in the experiment. A total force of 10g was applied to the rat molars to induce tooth movement. A Ga-Al-As diode laser was used to irradiate the area around the moving tooth and, after 7 days, the amount of tooth movement was measured. To determine the amount of tooth movement, plaster models of the maxillae were made using a silicone impression material before (day 0) and after tooth movement (days 1, 2, 3, 4, and 7). The models were scanned using a contact-type three-dimensional (3-D) measurement apparatus. Immunohistochemical staining for MMP-9, cathepsin K, and integrin subunits of alpha(v)beta3 was performed. Intergroup comparisons of the average values were conducted with a Mann-Whitney U-test for tooth movement and the number of tartrate-resistant acid phosphatase (TRAP), MMP-9, cathepsin K, and integrin subunits of alpha(v)beta3-positive cells. In the laser-irradiated group, the amount of tooth movement was significantly greater than that in the non-irradiated group at the end of the experiment (P < 0.05). Cells positively stained with TRAP, MMP-9, cathepsin K, and integrin subunits of alpha(v)beta3 were found to be significantly increased in the irradiated group on days 2-7 compared with those in the non-irradiated group (P < 0.05). These findings suggest that low-energy laser irradiation facilitates the velocity of tooth movement and MMP-9, cathepsin K, and integrin subunits of alpha(v)beta3 expression in rats.

Low-energy laser irradiation accelerates the velocity of tooth movement via stimulation of the alveolar bone remodeling

INTRODUCTION Previously, the authors have reported the acceleration of tooth movement and osteoclastogenesis on the pressure site in an experimental tooth movement model by low-energy laser irradiation (LELI), which stimulated the RANK/RANKL system and c-fms/macrophage colony-stimulating factor system. However, the effect of LELI on osteogenesis on the tension site is not known clearly. Moreover, the temporal changes in alveolar bone during tooth movement have not been investigated as yet. Therefore, the present study was designed to examine the effects of LELI on alveolar bone remodeling during experimental tooth movement, and observe the temporal bone mineral density (BMD) using micro-computed tomography (muCT). MATERIALS AND METHODS To induce experimental tooth movement in rats, 10 g force was applied to the upper right first molar with Nickel titanium closed-coil. Next, a gallium-aluminum-arsenide (Ga-Al-As) diode laser was used to irradiate the area around the moved tooth, and BMD and the amount of tooth movement were measured by muCT scanning for 21 days. Histopathological examination was also performed. RESULTS The amount of tooth movement in the LELI group was significantly greater than in the non-irradiation group by the end of the experimental period. Further, compared with the non-irradiation group, the fall of BMD was less in the LELI group. CONCLUSION These findings suggest that LELI accelerates the velocity of tooth movement via stimulation of the alveolar bone remodeling.

The tyrosine phosphatase CD148 interacts with the p85 regulatory subunit of phosphoinositide 3-kinase.

CD148 is a transmembrane tyrosine phosphatase that has been implicated in the regulation of cell growth and transformation. However, the signalling mechanisms of CD148 are incompletely understood. To identify the specific intracellular molecules involved in CD148 signalling, we carried out a modified yeast two-hybrid screening assay. Using the substrate-trapping mutant form of CD148 (CD148 D/A) as bait, we recovered the p85 regulatory subunit of PI3K (phosphoinositide 3-kinase). CD148 D/A, but not catalytically active CD148, interacted with p85 in a phosphorylation-dependent manner in vitro and in intact cells. Growth factor receptor and PI3K activity were also trapped by CD148 D/A via p85 from pervanadate-treated cell lysates. CD148 prominently and specifically dephosphorylated p85 in vitro. Co-expression of CD148 reduced p85 phosphorylation induced by active Src, and attenuated the increases in PI3K activity, yet CD148 did not alter the basal PI3K activity. Finally, CD148 knock-down by siRNA (short interfering RNA) increased PI3K activity on serum stimulation. Taken together, these results demonstrate that CD148 may interact with and dephosphorylate p85 when it is phosphorylated and modulate the magnitude of PI3K activity.

IL-8 and MCP-1 induced by excessive orthodontic force mediates odontoclastogenesis in periodontal tissues

OBJECTIVE The aim of this study was to investigate how interleukin (IL)-8 (cytokine-induced neutrophil chemoattractant; CINC-1) and monocyte chemotactic protein (MCP)-1/CCL2 contribute to root resorption during orthodontic tooth movement. MATERIALS AND METHODS Forty 6-week-old male Wistar rats were subjected to orthodontic force of 10 or 50 g to induce a mesially tipping movement of the upper first molars for 7 days. We determined the expressions of CINC-1, CXCR2, and MCP-1 proteins in root resorption area using immunohistochemistry. Furthermore, we investigated the effects of compression forces (CF) on IL-8 and MCP-1 production by human periodontal ligament (hPDL) cells. We observed an effect of chemokine treatment on rat odonto/osteoclasts in dentin slices that recapitulated root resorption. RESULTS The immunoreactivity for CINC-1/CXCR2 and MCP-1 was detected in odontoclasts and PDL fibroblasts by the orthodontic force of 50 g on day 7. CF increased the secretion and the expression of mRNA of IL-8 and MCP-1 from PDL cells in a magnitude-dependent manner. Moreover, CINC-1 and MCP-1 stimulated osteoclastogenesis from rat osteoclast precursor cells. CONCLUSION IL-8 (CINC-1) and MCP-1 may therefore facilitate the process of root resorption because of excessive orthodontic force.

Magnetic resonance imaging of osteomyelitis in the mandible: Comparative study with other radiologic modalities

Magnetic resonance imaging of 14 histopathologically confirmed cases of osteomyelitis of the mandible wasretrospectively reviewed. The findings of magnetic resonance imaging were compared with conventional radiography, computed tomography, bone scintigraphy, and histopathologic examinations. All lesions in bone marrow were shown as areas of low (64%) or low-to-intermediate (36%) signal intensity on T1-weighted images, and areas of high (29%), mixed (high and low, 21%; high and intermediate, 36%) or low (14%) signal intensity on T2-weighted images. Histopathologically, high T2-weighted signal intensity areas that showed enhancement after contrast injection corresponded to active infection. These were not collections of pus but were predominantly areas of granulation tissue. Magnetic resonance imaging showed larger areas of abnormality than plain radiography or computed tomography. Bone scintigraphy did not accurately reveal the locations of lesions but showed heterogeneous increased uptake in all patients. MRI was an extremely useful technique for assessing osteomyelitis of the mandible.

Low-energy laser irradiation stimulates the tooth movement velocity via expression of M-CSF and c-fms

Abstract Recent studies have demonstrated that low-energy laser irradiation stimulates bone formation in vitro and in vivo. However, very little is known about the effects of laser irradiation on osteoclastogenesis. Macrophage colony-stimulating factor (M-CSF) is essential and sufficient for osteoclastogenesis. The present study was designed to examine the effects of low-energy laser irradiation on expressions of M-CSF and c-fms during experimental tooth movement. A total of 10 g of orthodontic force was applied to rat molars to cause experimental tooth movement. A Ga-Al-As diode laser was used to irradiate the area around the moved tooth and the amount of tooth movement was measured for 7 days. Immunohistochemical staining with M-CSF and c-fms was performed. RT-PCR was also performed to elucidate the expression of c-fms from irradiated rat osteoclast precursor cells. In the irradiation group, the amount of tooth movement was significantly greater than that of the non-irradiation group at the end of the experiment. Cells positively stained with M-CSF and c-fms were found to be significantly increased in the irradiation group on days 2 and 3 as compared with the non-irradiation group. Further, c-fms expression in osteoclast precursor cells was detected at an early stage (days 2 and 3) in the irradiation group. These findings suggest that low-energy laser irradiation stimulates the velocity of tooth movement via the expressions of M-CSF and c-fms.

Kaempferol stimulates bone sialoprotein gene transcription and new bone formation

Kaempferol is a typical flavonol‐type flavonoid that is present in a variety of vegetables and fruits, and has a protective effect on postmenopausal bone loss. Bone sialoprotein (BSP) is thought to function in the initial mineralization of bone and could be crucial for osteoblast differentiation, bone matrix mineralization and tumor metastasis. In the present study we investigated the regulation of BSP transcription by kaempferol in rat osteoblast‐like UMR106 cells, and the effect of kaempferol on new bone formation. Kaempferol (5 µM) increased BSP and Osterix mRNA levels at 12 h and up‐regulated Runx2 mRNA expression at 6 h. Kaempferol increased luciferase activity of the construct pLUC3, which including the promoter sequence between nucleotides −116 to +60. Transcriptional stimulation by kaempferol abrogated in constructs included 2 bp mutations in the inverted CCAAT, CRE, and FRE elements. Gel shift analyses showed that kaempferol increased nuclear protein binding to CRE and FRE elements, whereas the CCAAT‐protein complex did not change after kaempferol stimulation. Twelve daily injections of 5 µM kaempferol directly into the periosteum of parietal bones of newborn rats increased new bone formation. These data suggest that kaempferol increased BSP gene transcription mediated through inverted CCAAT, CRE, and FRE elements in the rat BSP gene promoter, and could induce osteoblast activities in the early stage of bone formation. J. Cell. Biochem. 110: 1342–1355, 2010.

T-helper 17 cells mediate the osteo/odontoclastogenesis induced by excessive orthodontic forces

OBJECTIVE The aim of this study was to investigate how T-helper 17 cells (Th17 cells), interleukin (IL)-17, and interleukin-6 contribute to root resorption during orthodontic tooth movement. MATERIALS AND METHODS Fifteen male 6-week-old Wistar rats were subjected to orthodontic force of 10 or 50 g to induce a mesially tipping movement of the upper first molars for 7 days. The expression levels of TRAP, IL-17, the IL-17 receptor (IL-17R), and IL-6 proteins were determined in periodontal ligament (PDL) by immunohistochemical analysis. Moreover, the fluorescent localization immunoassay was performed to detect Th17 cells. Furthermore, the effects of IL-17 on IL-6 release were investigated using human PDL cells in vitro. The effect of IL-17 on osteoclastogenesis was evaluated by TRAP staining, actin ring staining, and the pit formation assay. RESULTS The immunoreactivity for Th17, IL-17, IL-17R, and IL-6 was detected in PDL tissue subjected to the orthodontic force on day 7. IL-17 increased the release of IL-6 from human periodontal ligament cells in a time-dependent manner. Moreover, IL-17 stimulated osteoclastogenesis from human osteoclast precursor cells, and these effects were partially suppressed by an anti-IL-6 antibody. CONCLUSION These results suggest that Th17 cells may aggravate the process of orthodontically induced inflammatory root resorption.

Cellular schwannoma in the oral mucosa

A case of cellular schwannoma of the oral mucosa in a 34-year-old Japanese man is described. Cellular schwannoma commonly affects soft tissues such as the retroperitoneum and posterior mediastinum, and also bone, but is extremely rare in the oral region. To our knowledge, this is only the second report of oral cellular schwannoma. Histologically, the tumor parenchyma consisted of hypercellular spindle cells with nuclear and cytoplasmic pleomorphism and nuclear palisading resembling Antoni A-type conventional schwannoma, without evidence of Verocay bodies. These features were indicative of cellular schwannoma. Immunohistochemically, the tumor cells were positive for S-100, S-100alpha, S-100beta and vimentin, suggesting that they were of peripheral nervous origin. Furthermore, it is speculated that the tumor was intermediate between a benign and a malignant state, based on the histological features and positivity for S-100alpha.