研究者詳細 - 長塚　仁

Oral Squamous Cell Carcinoma-derived Sonic Hedgehog Promotes Angiogenesis 国際誌

Hiromasa Kuroda, Naito Kurio, Tsuyoshi Shimo, Kenichi Matsumoto, Masanori Masui, Kiyofumi Takabatake, Tatsuo Okui, Soichiro Ibaragi, Yuki Kunisada, Kyoichi Obata, Norie Yoshioka, Koji Kishimoto, Hitoshi Nagatsuka, Akira Sasaki

Anticancer Research 37 ( 12 ) 6731 - 6737 2017年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：INT INST ANTICANCER RESEARCH

Background: Sonic hedgehog (SHH) signaling is related to the pathogenesis of oral squamous cell carcinoma (OSCC), but its role in OSCC is not yet well understood. In this study, we analyzed the role of SHH signaling in OSCC. Materials and Methods: We examined the expression pattern of SHH and its signal proteins in clinically resected OSCC samples by immunohistochemistry. We also evaluated the function of SHH signaling using the hedgehog signaling inhibitor cyclopamine in vivo and in vitro by proliferation, migration and angiogenesis analyses. Results: We found that SHH was highly expressed in human tongue OSCC, whereas patched (PTCH1), glioma-associated oncogene 1 (GLI1) and GLI2 proteins were expressed in the microvascular cells in the tumor invasive front. Administration of cyclopamine to mice suppressed the growth and angiogenesis of OSCC xenografts in vivo. Moreover, cyclopamine inhibited endothelial cell proliferation and migration, and reduced aorta vascular length in the rat. Conclusion: These findings suggest that OSCC-derived SHH stimulates angiogenesis at the tumor invasive front.

DOI： 10.21873/anticanres.12132

Antibacterial activity and biocompability of zinc oxide and graphite particles as endodontic materials

Silvia Noemí Kozuszko, María Alejandra Sánchez, María Inés Gutiérrez de Ferro, Ana María Sfer, Ana Paula Moreno Madrid, Kiyofumi Takabatake, Keisuke Nakano, Hitoshi Nagatsuka, Andrea Paola Rodríguez

Journal of Hard Tissue Biology 26 ( 4 ) 311 - 318 2017年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：JOURNAL HARD TISSUE BIOLOGY

The aim of this work is to evaluate the antibacterial effect and biocompatibility of zinc oxide (ZnO) nanopaticles and graphite-type carbon (Gt) microparticles commercial powders. SEM analysis was performed to assess particles morphology. The antibacterial behavior was studied against Staphylococcus aureus (Staph. Aureus) (bacterial strain ATCC 29213) and TEM analysis of bacteria was performed to determine ultrastructural alterations; in addition, biocompatibility was evaluated in subcutaneous tissue of Wistar rats at 3, 7 and 28 d. ZnO and Gt powders exhibited antibacterial activity while TEM images of Staph. aureus showed membrane disruption followed by the release of internal content. Also, an electron-light region within the cytoplasm was observed for microorganisms treated with ZnO. Regarding biocompatibility, Gt samples induced a foreign body reaction response with presence of giant cells whereas ZnO samples showed fibroblastic connective tissue with chronic inflammatory cells and new small vessels. Also, collagen fibers and lack of capsule was observed by Trichome Masson stain. Thus, ZnO improved wound healing by enhancing tissue regeneration in contrast with calcium hydroxide control sample response which showed a fibrous tissue scar. Hence, ZnO nano-powder seems to be a potential material in the regenerative endodontic field.

DOI： 10.2485/jhtb.26.311

GFPマウス骨髄移植モデルの異所性骨形成における骨髄由来細胞の役割(The role of bone marrow-derived cells in ectopic bone regeneration in the GFP mouse bone marrow transplantation model)

Tachykinin receptor 3 distribution in human oral squamous cell carcinoma 国際誌

Kyoichi Obata, Tsuyoshi Shimo, Tatsuo Okui, Kenichi Matsumoto, Hiroyuki Takada, Kiyofumi Takabatake, Yuki Kunisada, Soichiro Ibaragi, Hitoshi Nagatsuka, Akira Sasaki

Anticancer Research 36 ( 12 ) 6335 - 6341 2016年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：INT INST ANTICANCER RESEARCH

Background: Tachykinin 3 (TAC3) and its preferred tachykinin receptor 3 (TACR3) that are prominently detected in the central nervous system, play significant roles in physiological development and specifically in the human reproductive system. The roles of TAC3/TACR3 in oral squamous cell carcinoma are unknown. Materials and Methods: We examined the expression pattern of TAC3/TACR3 in clinically-resected oral squamous cell carcinoma samples using immunohistochemistry and immunofluorescence analysis. Results: We found that even though the expression level of TACR3 was negative in the normal epithelium, it was highly elevated in tumor cells. A more intense signal was observed in the invasive front of tumor cells that had migrated into the mandible bone matrix. TAC3 was not detected in tumor cells, but was expressed in PGP-9.5-positive sensory nerves in the mandible. Conclusion: Our results suggest that peripheral sensory nerve-derived TAC3 may affect gingival oral squamous cell carcinoma cells through TACR3 in the bone matrix.

DOI： 10.21873/anticanres.11230

Yu Song, Takabatake Kiyofumi, Tsujigiwa Hidetsugu, Matsuda Hiroyuki, Kawai Hotaka, Yoshida Saori, Nakano Keisuke, Nagatsuka Hitoshi

Journal of Hard Tissue Biology 25 ( 4 ) 458 - 458 2016年10月

エナメル上皮腫において間質特性をもたらすCCN2とYAP局在化の解析(Analyzing the localization of CCN2 and YAP which effect the stromal properties in ameloblastoma)

Hamada Mei, Takabatake Kiyofumi, Tsujigiwa Hidetsugu, Fujii Masae, Ono Sawako, Nobunaga Hikari, Nakano Keisuke, Nagatsuka Hitoshi

Journal of Hard Tissue Biology 25 ( 4 ) 460 - 461 2016年10月

Invadopodia proteins, cortactin, N-WASP and WIP differentially promote local invasiveness in ameloblastoma 国際誌

Chong Huat Siar, Zainal Ariff Bin Abdul Rahman, Hidetsugu Tsujigiwa, Kamila Mohamed Om Alblazi, Hitoshi Nagatsuka, Kok Han Ng

Journal of Oral Pathology and Medicine 45 ( 8 ) 591 - 598 2016年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：WILEY-BLACKWELL

Background: Cell migration and invasion through interstitial tissues are dependent upon several specialized characteristics of the migratory cell notably generation of proteolytic membranous protrusions or invadopodia. Ameloblastoma is a benign odontogenic epithelial neoplasm with a locally infiltrative behaviour. Cortactin and MMT1-MMP are two invadopodia proteins implicated in its local invasiveness. Other invadopodia regulators, namely N-WASP, WIP and Src kinase remain unclarified. This study addresses their roles in ameloblastoma. Materials and method: Eighty-seven paraffin-embedded ameloblastoma cases (20 unicystic, 47 solid/multicystic, 3 desmoplastic and 17 recurrent) were subjected to immunohistochemistry for expression of cortactin, N-WASP, WIP, Src kinase and F-actin, and findings correlated with clinicopathological parameters. Results: Invadopodia proteins (except Src kinase) and F-actin were widely detected in ameloblastoma (cortactin: n = 73/87, 83.9%; N-WASP: n = 59/87; 67.8%; WIP: n = 77/87; 88.5%; and F-actin: n = 87/87, 100%). Protein localization was mainly cytoplasmic and/or membranous, and occasionally nuclear for F-actin. Cortactin, which functions as an actin-scaffolding protein, demonstrated significantly higher expression levels within ameloblastoma tumoral epithelium than in stroma (P < 0.05). N-WASP, which coordinates actin polymerization and invadopodia-mediated extracellular matrix degradation, was overexpressed in the solid/multicystic subtype (P < 0.05). WIP, an upstream regulator of N-WASP, and F-actin were significantly upregulated along the tumour invasive front compared to tumour centres (P < 0.05). Except for males with cortactin overexpression, other clinical parameters (age, ethnicity and anatomical site) showed no significant correlations. Conclusions: Present results suggest that local invasiveness of ameloblastoma is dependent upon the migratory potential of its tumour cells as defined by their distribution of cortactin, N-WASP and WIP in correlation with F-actin cytoskeletal dynamics.

DOI： 10.1111/jop.12417

A case of a calcifying cystic odontogenic tumor with odontoma in a 5-year-old boy

Norifumi Moritani, Naoki Nakata, Eiki Yamachika, Tatsushi Matsumura, Hitoshi Nagatsuka, Seiji Iida

Journal of Oral and Maxillofacial Surgery, Medicine, and Pathology 28 ( 5 ) 421 - 425 2016年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Elsevier Ltd

We report a case of calcifying cystic odontogenic tumor (CCOT) with odontoma developing in the deciduous dentition period, and present previously reported cases in Japan. A 5-year-old boy visited a dental clinic because of swelling on his left mandibular buccal alveolar region, and was referred to our department. Clinical examination revealed a 20-mm (in diameter), bone-like, hard mass on the left mandibular buccal gingiva at the first deciduous molar region. A panoramic radiograph showed a 25 mm × 20 mm, unilocular, cystic radiolucent area from the left mandibular second deciduous molar to the first molar region. There were radiopaque tiny flecks at the upper side of the lesion. The clinical diagnosis was a benign tumor of the left mandible. Consequently, tumor enucleation was performed under general anesthesia. Histopathological diagnosis confirmed a CCOT associated with odontoma. Five years after surgery, the left mandibular region has healed well, with no evidence of recurrence.

DOI： 10.1016/j.ajoms.2016.03.009

移植骨髄由来細胞の口蓋腺への遊走と細胞分化

大谷友加里, 玉村亮, 河野哲朗, 鈴木久仁博, 中野敬介, 長塚仁, 辻極秀次, 岡田裕之

Journal of Oral Biosciences Supplement 2016 352 - 352 2016年9月

Three-Dimensional Cell Culture at the Frontiers of in Vitro Cancer Research: Present Perspectives

Soo LL, Zainal AA, Hidetsugu T, Takabatake K, Nakano K, Chai WL, Nagatsuka H, Siar CH

Annals of Dentistry 23 ( 1 ) 1 - 10 2016年6月

Efficacy of honeycomb TCP-induced microenvironment on bone tissue regeneration in craniofacial area 国際誌

Satoko Watanabe, Kiyofumi Takabatake, Hidetsugu Tsujigiwa, Toshiyuki Watanabe, Eijiro Tokuyama, Satoshi Ito, Hitoshi Nagatsuka, Yoshihiro Kimata

International Journal of Medical Sciences 13 ( 6 ) 466 - 476 2016年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：IVYSPRING INT PUBL

Artificial bone materials that exhibit high biocompatibility have been developed and are being widely used for bone tissue regeneration. However, there are no biomaterials that are minimally invasive and safe. In a previous study, we succeeded in developing honeycomb β-tricalcium phosphate (β-TCP) which has through-and-through holes and is able to mimic the bone microenvironment for bone tissue regeneration. In the present study, we investigated how the difference in hole-diameter of honeycomb β-TCP (hole-diameter: 75, 300, 500, and 1600 μm) influences bone tissue regeneration histologically. Its osteoconductivity was also evaluated by implantation into zygomatic bone defects in rats. The results showed that the maximum bone formation was observed on the β-TCP with hole-diameter 300μm, included bone marrow-like tissue and the pattern of bone tissue formation similar to host bone. Therefore, the results indicated that we could control bone tissue formation by creating a bone microenvironment provided by β-TCP. Also, in zygomatic bone defect model with honeycomb β-TCP, the result showed there was osseous union and the continuity was reproduced between the both edges of resected bone and β-TCP, which indicated the zygomatic bone reproduction fully succeeded. It is thus thought that honeycomb β-TCP may serve as an excellent biomaterial for bone tissue regeneration in the head, neck and face regions, expected in clinical applications.

DOI： 10.7150/ijms.15560

In Vivo evaluation of adipogenic induction in fibrous and honeycomb-structured atelocollagen scaffolds 国際誌

Andrea P. Rodríguez, Betiana Felice, María A. Sánchez, Hidetsugu Tsujigiwa, Carmelo J. Felice, Hitoshi Nagatsuka

Materials Science and Engineering C 63 125 - 130 2016年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ELSEVIER SCIENCE BV

Nowadays, soft tissue restoration techniques are mainly focused on volume regeneration instead of function recovering. So far, autologous fat transplant has been the most popular method although its multiple reported problems like volume and function loss. Adipose tissue engineering therefore emerges as a solution for development of biological substitutes for soft tissue which promotes not only volume regeneration but also function restoration with minimal consequences. Here we tested fibrous-structured atelocollagen (FSA) scaffolds and honeycomb atelocollagen (HCA) scaffolds for their ability to induce adipogenesis in vivo. Implants were subjected to histological and immunohistochemical assessment after 1, 2, and 4 weeks of implantation. Our studies showed that FSA scaffolds induced in vivo a markedly adipogenic response, whereas an acute inflammatory process was observed at HCA scaffolds without tissue regeneration detected within them. Our histological findings concerning FSA scaffolds clearly showed the presence of adipose-like tissue surprisingly composed by a mixture of brown-like and white-like adipocytes at week 2 whereas only white-like adipocytes at week 4. Subsequent positive Pax7 immunostaining at weeks 1 and 2 suggested the existence of a common myogenic progenitor shared by brown-like and white-like adipocytes observed. Then, in this work we present FSA scaffolds as a promising structure for brown and white adipose tissue engineering.

DOI： 10.1016/j.msec.2016.02.055

Novel midkine inhibitor iMDK inhibits tumor growth and angiogenesis in oral squamous cell carcinoma 国際誌

Masanori Masui, Tatsuo Okui, Tsuyoshi Shimo, Kiyofumi Takabatake, Takuya Fukazawa, Kenichi Matsumoto, Naito Kurio, Soichiro Ibaragi, Yoshio Naomoto, Hitoshi Nagatsuka, Akira Sasaki

Anticancer Research 36 ( 6 ) 2775 - 2781 2016年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：INT INST ANTICANCER RESEARCH

Midkine is a heparin-binding growth factor highly expressed in various human malignant tumors. However, its role in the growth of oral squamous cell carcinoma is not well understood. In this study, we analyzed the antitumor effect of a novel midkine inhibitor (iMDK) against oral squamous cell carcinoma. Administration of iMDK induced a robust antitumor response and suppressed cluster of differentiation 31 (CD31) expression in oral squamous cell carcinoma HSC-2 cells and SAS cells xenograft models. iMDK inhibited the proliferation of these cells dose-dependently, as well as the expression of midkine and phospho-extracellular signal-regulated kinase in HSC-2 and SAS cells. Moreover, iMDK significantly inhibited vascular endothelial growth factor and induced tube growth of human umbilical vein endothelial cells in a dose-dependent fashion. These findings suggest that midkine is critically involved in oral squamous cell carcinoma and iMDK can be effectively used for the treatment of oral squamous cell carcinoma.

The role of sonic hedgehog signaling in osteoclastogenesis and jaw bone destruction 国際誌

Tsuyoshi Shimo, Kenichi Matsumoto, Kiyofumi Takabatake, Eriko Aoyama, Yuichiro Takebe, Soichiro Ibaragi, Tatsuo Okui, Naito Kurio, Hiroyuki Takada, Kyoichi Obata, Pai Pang, Masahiro Iwamoto, Hitoshi Nagatsuka, Akira Sasaki

PLoS ONE 11 ( 3 ) e0151731 2016年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：PUBLIC LIBRARY SCIENCE

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.

DOI： 10.1371/journal.pone.0151731

Anti-osteoporosis effects of 1,4-dihydroxy-2-naphthoic acid in ovariectomized mice with increasing of bone density

Kenichiro Kita, Eiki Yamachika, Masakazu Matsubara, Hidetsugu Tsujigiwa, Nobuhisa Ishida, Norifumi Moritani, Tatsushi Matsumura, Masahide Mizutani, Yuki Fujita, Kiyofumi Takabatake, Kenichiro Ejima, Hitoshi Nagatsuka, Yoshinori Yamaguchi, Seiji Iida

Journal of Oral and Maxillofacial Surgery, Medicine, and Pathology 28 ( 1 ) 66 - 72 2016年1月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Elsevier Ltd

Objective: 1,4-Dihydroxy-2-naphthoic acid (DHNA) is the main component of the metabolic products after fermentation by Propionibacterium freudenreichii ET-3. In this study, DHNA, vitamin K2 (VK2), and risedronate (Ris) were administered to ovariectomized (OVX) mice to ascertain their suppressive effects on bone mass reduction. Methods: Fifty mice were divided into five groups: Sham, OVX, DHNA, VK2, and Ris groups. The drugs were administered daily for 8 weeks. Subsequently, blood was collected from the orbital venous plexus. The bilateral femurs and L5 lumbar vertebra were excised. To determine the effects of the drugs, we performed histomorphometric analyses of the left femur and L5 by micro-CT, biochemical analyses of serum samples, and histological analyses of the right femur. Results: The results for the total bone mineral density, bone volume density, trabecular thickness, trabecular number, trabecular separation, and histological analyses showed that bone resorption was improved in the DHNA and Ris groups compared with the OVX group. Furthermore, the biochemical analyses revealed that the production of inflammatory cytokines was suppressed in the DHNA and Ris groups. Conclusions: The present findings show that DHNA suppresses the bone mass reduction in OVX mice, suggesting that it could be an alternative treatment for postmenopausal osteoporosis.

DOI： 10.1016/j.ajoms.2015.07.001

Surgical treatment and dental implant rehabilitation after the resection of an osseous dysplasia

Shintaro Sukegawa, Takahiro Kanno, Hotaka Kawai, Akane Shibata, Kenichi Matsumoto, Yuka Sukegawa-Takahashi, Kyosuke Sakaida, Hitoshi Nagatsuka, Yoshihiko Furuki

Journal of Hard Tissue Biology 25 ( 4 ) 437 - 441 2016年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：JOURNAL HARD TISSUE BIOLOGY

Osseous dysplasia (OD), which is subdivided into four subtypes (focal cement-osseous dysplasia, florid osseous dysplasia, periapical cemental dysplasia, and familial gigantiform cementoma), is an idiopathic process located in the periapical region of the tooth-bearing jaw areas, characterized by a replacement of normal bone by fibrous tissue and metaplastic bone. Unless accompanied by bulging or secondary infection of the jawbone, treatment is not necessary. However, treatment for extirpation is required when a secondary infection is present. Consequently, occlusion reconstruction becomes difficult because of large bone defect. Herein, we report the surgical technique to maintain the alveolar ridge form after resecting the lesion and for the case of an infected alveolar bone in a patient with OD. The loss of the buccal cortical bone was inevitable after removal of the infected area. For postoperative occlusal reconstruction, we performed a bone graft to maintain the alveolar ridge form at the same time as the tumor extirpation. Deficient buccal cortical bone was rebuilt with bone taken from the mandibular ramus and a bioactive resorbable plate. We describe the management of OD and the surgical technique for alveolar ridge form management by resecting the lesion and infected alveolar bone.

DOI： 10.2485/jhtb.25.437

A clinical retrospective study of surgical treatment for medication-related osteonecrosis of the jaw

Shintaro Sukegawa, Takahiro Kanno, Hotaka Kawai, Satoko Nakamura, Akane Shibata, Yuka Sukegawa-Takahashi, Hitoshi Nagatsuka, Yoshihiko Furuki

Journal of Hard Tissue Biology 25 ( 4 ) 447 - 454 2016年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：JOURNAL HARD TISSUE BIOLOGY

In 2014, the American Association of Oral and Maxillofacial Surgery recommended surgical treatment for medication-related osteonecrosis of the jaw (MRONJ) patients classified as stage 3 or those with mobile segments of bony sequestrum. However, there is limited information regarding the healing mechanism in MRONJ surgical treatments. This study aimed to retrospectively elucidate clinical outcomes of the surgical treatment of MRONJ patients. This study included 26 patients (7 men, 19 women; age: 42–92 years; mean ± standard deviation, 75.3 ± 11.7 years) who were classified as stage 3 or had mobile segments of bony sequestrum, and intake of the offending drug was ceased. The sequestrum was removed with surrounding vital bone, and segmental resection was performed in one patient with a pathological mandibular fracture. The outcome was classified into one of the following three categories: “Healing,” “Improvement,” “Unchanged.” and “Exacerbation.” The mean postoperative follow-up period was 16.6 months (3.0 –57.9 months), and complete healing was observed in 21 patients (80.8%), improvement of the lesion was observed in four patients (15.4%), and the outcome of one patient was unchanged (3.8%). The procedure described here may be recommended with relatively high clinical success rate for the patients with MRONJ requiring surgical treatment.

DOI： 10.2485/jhtb.25.447

Nasopalatine duct cyst associated with dental implant treatment: A case report 査読

Shintaro Sukegawa, Takahiro Kanno, Hotaka Kawai, Yuichiro Takebe, Akane Shibata, Yuka Takahashi, Hitoshi Nagatsuka, Yoshihiko Furuki

Oral and Maxillofacial Surgery Cases 1 ( 3 ) 38 - 41 2015年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Elsevier Inc

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.

DOI： 10.1016/j.omsc.2015.06.003