Sonic hedgehog (SHH) and its signaling have been identified in several human cancers, and increased levels of its expression appear to correlate with disease progression and metastasis. However, the role of SHH in bone destruction associated with oral squamous cell carcinomas is still unclear. In this study we analyzed SHH expression and the role played by SHH signaling in gingival carcinoma-induced jawbone destruction. From an analysis of surgically resected lower gingival squamous cell carcinoma mandible samples, we found that SHH was highly expressed in tumor cells that had invaded the bone matrix. On the other hand, the hedgehog receptor Patched and the signaling molecule Gli-2 were highly expressed in the osteoclasts and the progenitor cells. SHH stimulated osteoclast formation and pit formation in the presence of the receptor activator for nuclear factor-κB ligand (RANKL) in CD11b+ mouse bone marrow cells. SHH upregulated phosphorylation of ERK1/2 and p38 MAPK, NFATc1, tartrate-resistant acid phosphatase (TRAP), and Cathepsin K expression in RAW264.7 cells. Our results suggest that tumor-derived SHH stimulated the osteoclast formation and bone resorption in the tumor jawbone microenvironment.

背景 : Gangliocytic paraganglioma (GP) は, 十二指腸乳頭部に好発するまれな腫瘍で, 主として粘膜下に存在するため, 生検による確定診断は困難なことが多い. EUS-FNA は術前診断の有効な方法として期待されるが, その報告は少ない.

BACKGROUND Tumor parenchyma-stromal interactions affect the properties of tumors and their dynamics. Our group previously showed that secreted frizzled related protein (sFRP)-2 impairs bone formation and promotes bone invasion in ameloblastoma. However, the effects of the secreted growth factors CCN2, TGF-β, and BMP4 on stromal tissues in ameloblastoma remain unclear. MATERIALS AND RESULTS Thirty-five paraffin-embedded ameloblastoma cases, ameloblastoma-derived cell lines (AM-1), and primary cultures of ameloblastoma stromal fibroblasts (ASF) were used. Immunohistochemistry, MTT assay, Western blotting, and RT-PCR were performed on these samples. Parenchyma-stromal CCN2 overexpression correlated significantly with fibrous-type stroma, but not with myxoid-type stroma, suggesting a role of CCN2 in fibrosis (P < 0.05). Recombinant CCN2 induction of enhanced ASF proliferation in AM-1 medium supports this view. Conversely, BMP4 and TGF-β were expressed in myxoid-type fibroblasts, but little expression was found in parenchyma. RANKL-positive and CD68-positive stromal cell populations were significantly greater in myxoid-type tumor areas than in fibrous-type tumor areas, while a higher Ki-67 labeling index was recorded in ameloblastoma with fibrous-type stroma. These data suggest that stromal properties influence bone resorption-related activities and growth rates, respectively. CONCLUSIONS These results suggest that the effects of secreted growth factors are governed by ameloblastoma parenchyma-stromal interactions. CCN2 promotes fibrogenesis independent of TGF-β signaling. Absence of CCN2 expression is associated with a phenotypic switch to a myxoid-type microenvironment that is conducive for TGF-β/BMP4 signaling to promote osteoclastogenesis.

INTRODUCTION: Immunoglobulin G4-related disease (IgG4-RD) is an inflammatory condition associated with elevated serum IgG4 levels and tissue infiltration by IgG4-expressing plasma cells. We present a case of adenoid cystic carcinoma (ACC) of the submandibular gland with possible involvement of IgG4-RD.PRESENTATION OF CASE: The patient was a 59-year-old man presenting with a swollen right submandibular gland. Laboratory tests revealed IgG4 levels of 176 mg/dl (reference range: 4.8-105). An initial open biopsy for histological diagnosis showed chronic sialadenitis. The region was monitored on an outpatient basis, and finally the right submandibular was totally resected because malignant tumor could not be excluded. Histological examination of the submandibular gland showed an ACC with lymphocytic infiltration containing many IgG4-positive plasma cells in the tumor stroma.DISCUSSION: We have described a case that indicated a possible involvement of ACC with IgG4-RD. This allows us to speculate that longstanding IgG4-RD may progress to malignancy or infiltration of IgG4-positive plasma cells through the signals of tumor stimuli. Further investigations are required to determine the potential pathogenic mechanism underlying this unique tumor.CONCLUSION: This case underscores that caution is needed in the diagnosis of masses with high serum IgG4 levels, as the differential diagnosis includes malignancy. (C) 2015 The Authors. Published by Elsevier Ltd. on behalf of Surgical Associates Ltd.

Maxillary anterior implants are associated with the risk of nasopalatine canal damage. Here we present the case of a 37-year-old man who developed a nasopalatine duct cyst after maxillary implant placement. The patient received an implant 3 months after the extraction of a fractured maxillary right central incisor. At a maintenance visit 9 years after the procedure, he complained of swelling and mild pain in the palatal region of the implant. A panoramic radiograph and computed tomography (CT) scan revealed a large, well-circumscribed, periapical radiolucency surrounding the apical portion of the implant and extending to the nasopalatine duct. We removed the entire lesion without removing the implant. Histopathologic examination of the resected specimen revealed a nasopalatine duct cyst. Accidental contact with the nasopalatine canal during implant surgery may have led to the development of the nasopalatine duct cyst. Careful planning using a preoperative CT scan prior to implant placement may prevent such complications.

Robotic-assisted surgery using the da Vinci Surgical System (Intuitive Surgical, Inc., Sunnyvale, California, USA) is an innovative procedure, and offers several advantages over conventional laparoscopy. These include wristed instruments, 3D vision, intuitive movements and tremor filtration. The shorter learning curve provided by this platform allows for effective training and widespread use of this minimally invasive approach to several operative methods. We aimed to clarify the effects of FIR ceramics activating the water and protein molecules on the metabolism activity of hatchery Tai fish by oral administration. The FIR ceramics were mixed with the food for hatchery Tai fish. The scale, muscle and liver were analyzed after 1 month. The Tai fish eating the food containing FIR ceramics grew quickly and got the resistance to red tide. The scale was absorbed and the muscle and liver were activated. Therefore, it was suggested that FIR ceramics activated blood circulation to improve the metabolism of hatchery Tai fish. II on the expression of osteogenesis-related transcription factors and extracellular matrix proteins in ROS17/2.8 cells. Runx2, Msx2, and osteocalcin expression significantly decreased with Ang II compared to the control, whereas AJ18 expression significantly increased. AT 1 blocker blocked suppressive or stimulatory effects of Ang II on Runx2, Msx2, osteocalcin, and AJ18 expression. These results suggest that Ang II suppresses osteoblastic differentiation by altering the expression of osteogenesis-related transcription factors via the AT 1 receptor and the function of osteogenesis in ROS17/2.8 cells. potential of HPV-16 for malignant transformation of OLP not been studied. In this study we assessed HPV-16 infection in Japanese OLP patients who developed cancer. HPV infection in 200 cases of OLP and 8 cases of OLP which developed cancer was detected by type specific PCR. The HPV-16 infection rate in 200 cases of OLP was 25.5% and in 8 cases of OLP which developed cancer was 62.5%. The results indicate that HPV-16 was often present in OLP and OLP which developed cancer. It suggests that HPV infection is a risk factor for malignant transformation in OLP. Our previous studies reported that nicotine doesn’t affect mineral resorption. In this study, we examined the direct effects of nicotine on the expression of bone resorption-related enzymes, actin organization, and bone resorption using RAW264.7 cells and bone marrow cells as osteoclast. Nicotine decreased the expression of cathepsin K, MMP-9 ， V-ATPase and the distribution of F-actin. Nicotine also suppressed the planar area of resorption pit on dentin slice by osteoclasts. These result indicate that nicotine reduce bone resorption by suppressing V-ATPase, cathepsin K and MMP-9 expression and actin organization. that Annexin a II (Anx a II) suppressed inflammatory disorders and thrombotic ischemic injuries in sepsis DIC through hyperfibrinolysis and anti-inflammatory. Anx a II may inhibit the thrombus formation through plasmin activation and suppression of cytokines like tissue factor (TF), but these mechanisms still remain unclear. To investigate we examined the influence of Anx a! and low molecular weight heparin (LMWH) thrombus formation and TF localization histopathologically, and plasma IL-6 expression,using ELISA in LPS-induced DIC model. The immunoreactivity of TF was negative on glomeruli without thrombus in Anx a II group, but positive on glomeruli without thrombus in LMWH group. IL-6 in Anx a II group reduced significantly. These results suggest that the Anx a II inhibits thrombus formation by not only plasmin activation,but also suppressing of the TF induced by IL-6. We examined the transplanted bone marrow-derived cell migration into periodontal tissues. Bone marrow cells from green fluorescent protein (GFP) transgenic mice were transplanted. This immunohistochemistry (IHC) revealed the GFP-positive cells were detected in the periodontal tissues. The GFP-positive cells histopathologically differentiated into some cell types. The fluorescence IHC and TRAP staining techniques demonstrated these cells were detected as macrophages and osteoclasts. Furthermore, GFP-positive cells gathered adjacent small blood vessels. These data suggest that GFP-positive bone marrow-derived cell migration into periodontal tissues and differentiate periodontal tissue component-cells. Analyze of variation of the tumor stroma by ameloblastoma cells. and oral squamous cell carcinoma(OSCC) with lymph node metastasis(LNM). for of HPV DNA, of underlying tubular damage caused by myoglobin-induced oxidative DNA damage is still unclear. We performed biochemical and histological analysis of myoglobin, malondialdehyde, 8-OHdG levels in the kidney tissue of a rat model of glycerol-induced rhabdomyolysis. The levels of myoglobin and malondialdehyde were found to increase at 3 hours after glycerol injection. Immunoreactivity for 8-OHdG was observed in the nuclei of proximal tubules at 0.5 hours. These results suggest that the proximal tubular injury by myoglobin may be caused through two different pathological processes, the cell membrane damage associated with the lipid peroxidation and the apoptosis associated with DNA oxidation. This study was designed to investigate the histopathological changes of the rat dental pulp directly-capped with a new light-curable resin-modified material containing calcium silicate and highly-hydrophilic monomer (TheraCal LC, BISCO, USA) comparing with those of MTA (ProRoot MTA, DENTSPLY-Sankin, Japan). In both experimental groups, slight chronic inflammatory cell infiltration was observed with almost complete or complete dentin-bridging at 7 days, while complete dentin bridge was formed without any inflammation at 14 days. This particular study revealed that the new light-curable direct pulp-capping agent containing calcium silicate possessed excellent ability of reparative healing of the exposed pulp, that was equivalent of MTA. We studied the proliferation and differentiation of osteoblastic cells derived from mesenchymal stem cell in diabetes model (SDT fatty) rat on titanium surface. The proliferation of osteoblastic cells was lower in the SDT group than in the control group at the initial stage of culture. The ALP activity of osteoblasitc cells was lower in the SDT group than in the control group at day14 cultures, however, that was no significant at day21 cultures. These results suggested that the osteoblastic cells of the SDT fatty rat have proliferative potential, differentiation potency and ability of calcification, these useful data which opened the research of the effects of diabetic to bone metabolism was provided. 40 cases of ameloblastoma using immunohistochemical techniques. To examine the relevance of HSP in cell differentiation, we focused on the cytoskeletal expression of CK. CK19 is a marker of typical odontogenic epithelium widely observed. Since staining with CK14 is one of the measures of the differentiation potential of squamous cells and is extensively expressed, it implies that squamous differentiation of each type can occur. CK8 was strongly detected in tumor nests in plexiform type but weakly detected in follicular type. The data suggested that HSP27 act as a possible molecular chaperon of cell differentiation in ameloblastomas. Notch signaling was examined ameloblastic fibroma cases using immunohistochemistry (IHC). There were positive products detected in the epithelial cells. But, the staining products of the mesemchymal fibroblastic cells were detected positively or negatively in case by case. The fibroblastic cells of odontogenic myxofibroma cases as control were also positive or negative to IHC. These data suggested that Notch act as a possible molecular chaperon of cell differentiation of odontogenic epithelial cell nests, although there are some differences of fibroblastic cells in the meschemcymal tissues among individual cases. We considered that the differences are according to the relationship between the epithelial and meschemcymal interactions in the neoplastic tissues.

Purpose of the research: Tooth germ development involves multiple events, including cell proliferation and cell differentiation. Connective tissue growth factor (CTGF/CCN2) is a signaling protein involved in tooth germ development, and we investigated how it is expressed and what roles it may have in primary cultures of mesenchymal cells derived from the developing tooth germ. We also examined the expression of CCN2 in a human odontogenic myxofibroma, a benign tumor of odontogenic mesenchymal origin, and considered the possible roles of CCN2 in the development of myxofibromas.Materials and methods: Mesenchymal cells of early bell-stage tooth germs were isolated from Day-90 bovine embryos and placed in primary culture. A resected specimen from a patient with odontogenic myxofibroma was prepared for immunohistochemical studies.Principal results: The CCN2 expression level in proliferating odontogenic mesenchymal cells freshly isolated from the early bell stage of developing bovine tooth germs and placed in primary culture was 3 times higher than that in the confluent non-proliferating cells. Recombinant CCN2 significantly increased the proliferation and type I collagen expression in odontogenic mesenchymal cells in primary culture. Immunohistochemical analysis on myxofibroma case revealed that CCN2 was detectable in MIB-1, a cellular marker of proliferation-positive odontogenic mesenchymal cells adjacent to capillary blood vessels and in the endothelial cells of the vessels in the tumor.Major conclusion: CCN2 signaling would influence the proliferation of and extracellular matrix production by dental mesenchymal cells. Our results suggest that the same mechanisms of CCN2 action toward dental mesenchymal cells would also be operative in the odontogenic myxofibroma microenvironment. c 2013 Japanese Stomatological Society. Published by Elsevier Ltd. All rights reserved.

Induced pluripotent stem (iPS) cells can be generated through the reprogramming of somatic cells by forced expression of defined exogenous factors. We previously reported that the gingiva, which is often resected during general dental treatments and treated as biomedical waste, is a promising source of iPS cells. These iPS cells were capable of terminal osteogenic differentiation, which could be greatly enhanced by small molecules obtained by a chemical library screening. In this presentation, generation and basic aspects of osteogenic capacity of the gingiva-derived iPS cells will be discussed, with an emphasis on potential applications of the iPS cell technologies to bone biology.

Tooth tissue engineering offers very attractive perspectives for elaboration of regenerative treatments, which enables to cure tooth loss and restore quality of life of the patients. To elaborate such treatment, isolation and culture of dental pulp cell must be achieved as a key element. In this article, we report the establishment of a stable cell line from GFP transgenic rat dental pulp, named TGC (Tooth Matrix-forming, GFP Rat-derived Cell). TGCs have exhibited odontoblastic feature both in vitro and in vivo. In vitro, TGC exposed to osteogenic medium demonstrated collagen fiber synthesis with matrix vesicle and mineralization and formed a sheet-like substrate on the cell culture dish. Increased ALP activity and elevated transcription level of various genes involved in calcification and dentin formation were also observed. In vivo, transplanted TGC in SCID mice with β-TCP particles formed dentin-like and pulp-like structure with lining odontoblast. Notably, even after up to 80 passages, TGCs retain their morphological features and differentiation ability. To our knowledge, this is the first report of a dental pulp-derived cell with such stable odontoblastic characteristics. TGC could be a very useful model for further study on dental pulp cell.