研究者詳細 - 長塚　仁

論文 - 長塚　仁

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ハニカムTCP構造の幾何学構造が骨・軟骨形成および血管新生に及ぼす影響(Effects of the geometrical structure of a honeycomb TCP structure on bone and cartilage formation and angiogenesis)

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

Journal of Hard Tissue Biology 27 ( 4 ) 367 - 367 2018年10月

口腔扁平上皮癌における腫瘍間質による腫瘍生物学特性の制御(Control of tumor biological character by tumor stroma in oral squamous cell carcinoma)

Takabatake Kiyofumi, Kawai Hotaka, Yoshida Saori, Matsuda Hiroyuki, Fujii Masae, Nakano Keisuke, Nagatsuka Hitoshi

Journal of Hard Tissue Biology 27 ( 4 ) 365 - 366 2018年10月

エナメル上皮腫における骨組織に対する実質-間質相互作用の影響(Effect of parenchyma-stromal interactions on bone tissue in ameloblastoma)

Sumi Ayaka, Sakaue Takaki, Hamada Mei, Takabatake Kiyofumi, Nagatsuka Hitoshi, Tsujigiwa Hidetsugu

Journal of Hard Tissue Biology 27 ( 4 ) 365 - 365 2018年10月

A case of sublingual adenoid cystic carcinoma involving the mandible presenting as a “skip lesion”

Mariko Fujita, Yoshinobu Yanagi, Arthur, R.G. Cortes, Emiko Saito Arita, Tomoo Onoda, Hitoshi Nagatsuka, un ichi Asaumi

Oral Radiology 34 ( 3 ) 281 - 287 2018年9月

Involvement of miR-140-3p in Wnt3a and TGFβ3 signaling pathways during osteoblast differentiation in MC3T3-E1 cells 査読国際誌

Shigeko Fushimi, Tsutomu Nohno, Hitoshi Nagatsuka, Hironobu Katsuyama

Genes to Cells 23 ( 7 ) 517 - 527 2018年7月

記述言語：英語掲載種別：研究論文（学術雑誌）

The Wnt/β-catenin signaling and TGFβ signaling pathways play a key role in osteoblast differentiation. The miRNAs play important roles in regulating gene expression at the post-transcriptional level through fine-tuning of protein-encoding gene expression. However, involvement of miRNAs is not established for Wnt3a and TGFβ signaling pathways in osteoblast differentiation. Here, we examined the role of miRNAs expressed differentially after Wnt3a expression during osteoblast differentiation. Over-expression of the Wnt3a gene increased ALP transcription, but decreased Col1, Runx2, and OCN transcription in osteoblastic MC3T3-E1 cells. Expression profiling and quantitative PCR for miRNAs showed that miR-140-3p decreased in Wnt3a-over-expressing osteoblastic cells. Wnt3a over-expression increased TGFβ3 expression, whereas transfection of the miR-140-3p mimic into MC3T3-E1 cells significantly inhibited TGFβ3 expression. Luciferase assay for the TGFβ3 transcript showed that TGFβ3 was a direct target of miR-140-3p. miR-140-3p mimic transfection resulted in significantly increased OCN transcription, but did not affect ALP, Col1, and Runx2 transcription in MC3T3-E1 cells. rTGFβ3 treatment decreased OCN transcription in MC3T3-E1 cells. These results suggest that the miR-140-3p is involved in osteoblast differentiation as a critical regulatory factor between Wnt3a and TGFβ3 signaling pathways.

DOI： 10.1111/gtc.12591

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肺癌移植マウスを用いた高転移臓器における骨髄由来細胞の局在と役割の検討

河合穂高, 辻極秀次, 信長ひかり, 高畠清文, 中野敬介, 長塚仁

日本がん免疫学会総会プログラム・抄録集 22回 97 - 97 2018年7月

Cell supplying to the experimentally induced absorbable suture thread foreign body granuloma from the bone marrow tissues

Y. Nakayasu, S. Aoki, M. Shoumura, N. Osuga, N. Okafuji, K. Nakano, H. Nagatsuka, H. Tsujigiwa, Toshiyuki Kawakami

International Journal of Dentistry and Oral Science 5 ( 6 ) 641 - 644 2018年6月

掲載種別：研究論文（学術雑誌）

Using the experimental system of GFP (Green florescence Protein) bone marrow transplanted rats, we implanted the Vicryl absorbable suture thread in subcutaneous tissue and examined the formation of foreign body granuloma histopathologically. We also compared the main cellular components, namely, the macrophage and foreign body giant cell immunohistochemically using GFP to confirm their origin. Histologically, the disintegrated suture thread was observed as a circular in void surrounded by the proliferation of macrophages and foreign body giant cells. Fibrous connective tissue including the fibroblasts was interposed among large masses. Even after 6 months, some clusters considered to be residues of macrophages that grew on the suture were still remained. Immunohistochemical examination of GFP showed that all proliferated macrophages and foreign body giant cells were GFP-positive. However, the relatively thin fibrous tissues formed on the outermost layer of the granulation tissue growth were mostly GFP-negative. Based on the results of the experiment, the macrophages and foreign body giant cells that are GFP-positive were derived from the mesenchymal tissue of transplanted bone marrow tissues.

DOI： 10.19070/2377-8075-18000126

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The role of bone marrow-derived cells during ectopic bone formation of mouse femoral muscle in GFP mouse bone marrow transplantation model 査読国際誌

Kiyofumi Takabatake, Hidetsugu Tsujigiwa, Yu Song, Hiroyuki Matsuda, Hotaka Kawai, Masae Fujii, Mei Hamada, Keisuke Nakano, Toshiyuki Kawakami, Hitoshi Nagatsuka

International Journal of Medical Sciences 15 ( 8 ) 748 - 757 2018年5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

Multipotential ability of bone marrow-derived cells has been clarified, and their involvement in repair and maintenance of various tissues has been reported. However, the role of bone marrow-derived cells in osteogenesis remains unknown. In the present study, bone marrow-derived cells during ectopic bone formation of mouse femoral muscle were traced using a GFP bone marrow transplantation model. Bone marrow cells from C57BL/6-Tg (CAG-EGFP) mice were transplanted into C57BL/6 J wild type mice. After transplantation, insoluble bone matrix (IBM) was implanted into mouse muscle. Ectopic bone formation was histologically assessed at postoperative days 7, 14, and 28. Immunohistochemistry for GFP single staining and GFP-osteocalcin double staining was then performed. Bone marrow transplantation successfully replaced hematopoietic cells with GFP-positive donor cells. Immunohistochemical analyses revealed that osteoblasts and osteocytes involved in ectopic bone formation were GFP-negative, whereas osteoclasts and hematopoietic cells involved in bone formation were GFP-positive. These results indicate that bone marrow-derived cells might not differentiate into osteoblasts. Thus, the main role of bone marrow-derived cells in ectopic osteogenesis may not be to induce bone regeneration by differentiation into osteoblasts, but rather to contribute to microenvironment formation for bone formation by differentiating tissue stem cells into osteoblasts.

DOI： 10.7150/ijms.24605

The prognostic implications of bone invasion in gingival squamous cell Carcinoma 査読国際誌

Shoko Yoshida, Tsuyoshi Shimo, Yurika Murase, Kiyofumi Takabatake, Koji Kishimoto, Soichiro Ibaragi, Norie Yoshioka, Tatsuo Okui, Hitoshi Nagatsuka, Akira Sasaki

Anticancer Research 38 ( 2 ) 955 - 962 2018年2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/Aim: This study evaluated the associations between bone invasion of gingival squamous cell carcinoma (SCC) and clinicopathological manifestations, and aimed to determine whether bone invasion is an independent prognostic factor in gingival SCC. Patients and Methods: The study was a retrospective review of 78 patients with gingival SCC who underwent surgery with curative intent. The level of bone invasion was pathologically categorized as medullary, cortical or no bone invasion. Results: Cortical and medullary bone invasion was present in 29 and 22 patients, respectively. There was a significant association between medullary bone invasion and tumor size (p=0.017), pathological N classification (p0.001), differentiation (p=0.017) and lymphovascular invasion (p=0.007). Medullary bone invasion and lymphovascular invasion were independent predictors of reduced overall survival (p=0.015, 0.048); medullary bone invasion was also an independent predictor of reduced disease-specific survival (p=0.018). Conclusion: Pathologically-proven medullary bone invasion and lymphovascular invasion were found to be key prognostic factors in gingival SCC. The results suggest that it is necessary to consider adjuvant therapy in patients with medullary bone invasion.

DOI： 10.21873/anticanres.12309

Characterization and potential roles of bone marrow-derived stromal cells in cancer development and metastasis 査読国際誌

Hotaka Kawai, Hidetsugu Tsujigiwa, Chong Huat Siar, Keisuke Nakano, Kiyofumi Takabatake, Masae Fujii, Mei Hamada, Ryo Tamamura, Hitoshi Nagatsuka

International Journal of Medical Sciences 15 ( 12 ) 1406 - 1414 2018年

記述言語：英語掲載種別：研究論文（学術雑誌）

Background: The tumor microenvironment and its stromal cells play an important role in cancer development and metastasis. Bone marrow-derived cells (BMDCs), a rich source of hematopoietic and mesenchymal stem cells, putatively contribute to this tumoral stroma. However their characteristics and roles within the tumor microenvironment are unclear. In the present study, BMDCs in the tumor microenvironment were traced using the green fluorescent protein (GFP) bone marrow transplantation model. Methods: C57BL/6 mice were irradiated and rescued by bone marrow transplantation from GFP-transgenic mice. Lewis lung cancer cells were inoculated into the mice to generate subcutaneous allograft tumors or lung metastases. Confocal microscopy, immunohistochemistry for GFP, α-SMA, CD11b, CD31, CD34 and CD105, and double-fluorescent immunohistochemistry for GFP-CD11b, GFP-CD105 and GFP-CD31 were performed. Results: Round and dendritic-shaped GFP-positive mononuclear cells constituted a significant stromal subpopulation in primary tumor peripheral area (PA) and metastatic tumor area (MA) microenvironment, thus implicating an invasive and metastatic role for these cells. CD11b co-expression in GFP-positive cells suggests that round/dendritic cell subpopulations are possibly BM-derived macrophages. Identification of GFP-positive mononuclear infiltrates co-expressing CD31 suggests that these cells might be BM-derived angioblasts, whereas their non-reactivity for CD34, CD105 and α-SMA implies an altered vascular phenotype distinct from endothelial cells. Significant upregulation of GFP-positive, CD31-positive and GFP/CD31 double-positive cell densities positively correlated with PA and MA (P<0.05). Conclusion: Taken together, in vivo evidence of traceable GFP-positive BMDCs in primary and metastatic tumor microenvironment suggests that recruited BMDCs might partake in cancer invasion and metastasis, possess multilineage potency and promote angiogenesis.

DOI： 10.7150/IJMS.24370

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Osteogenic capacity of mixed-acid and heat-treated titanium mesh prepared by a selective laser melting technique

Kayoko Yamamoto, Seiji Yamaguchi, Tomiharu Matsushita, Shigeo Mori, Azumi Hirata, Nahoko Kato-Kogoe, Hiroyuki Nakano, Yoichiro Nakajima, Yoshihiro Nishitani, Hitoshi Nagatsuka, Takaaki Ueno

RSC Advances 8 ( 46 ) 26069 - 26077 2018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：ROYAL SOC CHEMISTRY

The practical use of additive manufacturing to create artificial bone as a material for repairing complex bone defects is currently attracting attention. In this study, we compared the osteogenic capacity of materials composited by the method developed by Kokubo et al. of treating 3D-printed titanium (Ti) mesh with a mixture of H2SO4 and HCl and heating (mixed-acid and heat treatment) with that of materials subjected to conventional chemical treatment. Ti plates treated with this method have been found to promote highly active bone formation on their surface when inserted into rabbit tibial bone defects. No previous study has compared this method with other surface treatment methods. In this study, we used histological and other observations to compare the bone formation process in bone defects when Ti meshes prepared by the selective laser melting technique (SLM) and treated either with mixed acids and heat or with conventional chemical Ti surface treatments were implanted in a rat calvarial bone defect model. We found that both micro-computed tomography and observations of undecalcified ground sections showed that the best bone formation was observed in rats implanted with mesh treated with mixed acids and heat. Our results suggest that mixed-acid and heat-treated Ti mesh prepared by SLM may have a high osteogenic capacity in bone defects.

DOI： 10.1039/c8ra04193h

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DKK3 overexpression increases the malignant properties of head and neck squamous cell carcinoma cells 査読国際誌

Naoki Katase, Shin Ichiro Nishimatsu, Akira Yamauchi, Masahiro Yamamura, Kumiko Terada, Masumi Itadani, Naoko Okada, Nur Mohammad Monsur Hassan, Hitoshi Nagatsuka, Tohru Ikeda, Tsutomu Nohno, Shuichi Fujita

Oncology Research 26 ( 1 ) 45 - 58 2018年

記述言語：英語掲載種別：研究論文（学術雑誌）

DKK3, a member of the dickkopf Wnt signaling pathway inhibitor family, is believed to be a tumor suppressor because of its reduced expression in cancer cells. However, our previous studies have revealed that DKK3 expression is predominantly observed in head and neck/oral squamous cell carcinoma (HNSCC/OSCC). Interestingly, HNSCC/OSCC patients with DKK3 expression showed a high rate of metastasis and poorer survival, and siRNA-mediated knockdown of DKK3 in HNSCC-derived cancer cell lines resulted in reduced cellular migration and invasion. From these data, it was hypothesized that DKK3 might exert an oncogenic function specific to HNSCC. In the present research, the DKK3 overexpression model was established, and its influences were investigated, together with molecular mechanism studies. The DKK3 expression profile in cancer cell lines was investigated, including HNSCC/OSCC, esophageal, gastric, colorectal, pancreatic, prostatic, and lung cancers. DKK3 overexpression was performed in HNSCC-derived cells by transfection of expression plasmid. The effects of DKK3 overexpression were assessed on cellular proliferation, migration, invasion, and in vivo tumor growth. The molecular mechanism of DKK3 overexpression was investigated by Western blotting and microarray analysis. DKK3 overexpression significantly elevated cellular proliferation, migration, and invasion, as well as increased mRNA expression of cyclin D1 and c-myc. However, reporter assays did not show TCF/LEF activation, suggesting that the increased malignant property of cancer cells was not driven by the Wnt/β-catenin pathway. For the investigation of the pathways/molecules in DKK3-mediated signals, the Western blot analyses revealed that phosphorylation of Akt (S473) and c-Jun (Ser63) was elevated. The application of a PI3K kinase inhibitor, LY294002, on HSC-3 DKK3 cells significantly decreased tumor cell proliferation, migration, and invasion. From these results, we demonstrated that DKK3 might contribute to cellular proliferation, invasion, migration, and tumor cell survival in HNSCC cells through a mechanism other than the canonical Wnt signaling pathway, which might be attributed to PI3K-Akt signaling.

DOI： 10.3727/096504017X149268745963860965-0407

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In vitro efficacy of CaCO<inf>3</inf> content in CaTiO<inf>3</inf> - CaCO<inf>3</inf> composites for bone growth 査読

Andrea Paola Rodríguez, María Alejandra Sánchez, Betiana Felice, Martín Lucas Zamora, Hidetsugu Tsujigiwa, Kiyofumi Takabatake, Hotaka Kawai, Keisuke Nakano, Hitoshi Nagatsuka

Journal of Hard Tissue Biology 27 ( 3 ) 250 - 256 2018年

掲載種別：研究論文（学術雑誌）

The effect of CaTiO3 compounded with different amounts of CaCO3 on osteoblastic KUSA/A1 cells was evaluated. CaTiO3-CaCO3 composites were obtained by alkoxide method, a simple, low-cost and reproducible technique used for large-scale production of material. The content of CaCO3 in our samples was controlled by varying the sintering time of the overall process. Composite morphology was assessed by scanning electron microscopy (SEM) showing particles with sizes ranging from100 to 500 nm. The presence of CaCO3 was revealed by XRD and thermogravimetric analyses, which suggested that samples treated at 650ºC for 30 min contained higher amounts of CaCO3 than samples treated for 2 and 10 h. Additionally, in vitro studies demonstrated that CaTiO3–CaCO3 composites sintered for 30 min induced augmented cell proliferation and mineralization in comparison to composites sintered for longer periods of time. Hence, our findings clearly suggest that the amount of CaCO3 within CaTiO3-CaCO3 composites exerts a critical effect on osteoblastic cells response. Enhanced bone regeneration could be achieved by increasing the content of CaCO3 within the composites, thus establishing CaTiO3-CaCO3 as a promising material for bone augmentation procedures in dental field.

DOI： 10.2485/jhtb.27.250

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Role of neurokinin 3 receptor signaling in oral squamous cell carcinoma 国際誌

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

Anticancer Research 37 ( 11 ) 6119 - 6123 2017年11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/Aim: The neurokinin 3 receptor (NK-3R) is differentially expressed in the central nervous system including cases of human oral squamous cell carcinoma. However, the role of NK-3R signaling in oral squamous cell carcinoma is not well known. Materials and Methods: NK-3R expression in surgically resected oral squamous cell carcinoma was examined immunohistochemically and the strength of the expression was quantified. We evaluated the function of NK-3R signaling using NK-3R antagonist in human oral squamous cell carcinoma bone invasion mouse model. Results: NK-3R was significantly expressed in tumor cells that had invaded the bone matrix compared to the oral side tumor cells. SB222200, a selective antagonist of NK-3R, significantly suppressed the radiographic osteolytic lesion and tumorigenesis. Conclusion: NK-3R signaling is a potential target for the treatment of oral squamous cell carcinoma in cases of bone destruction.

DOI： 10.21873/anticanres.12060

腫瘍移植マウスを用いた、遠隔臓器に含まれる骨髄由来細胞の局在同定(Localization of bone marrow-derived cells of distant organs using tumor transplanted mouse)

Kawai Hotaka, Tsujigiwa Hidetsugu, Nobunaga Hikari, Takabatake Kiyofumi, Nakano Keisuke, Matsuda Hiroyuki, Fushimi Shigeko, Nagatsuka Hitoshi

Journal of Hard Tissue Biology 26 ( 4 ) 438 - 438 2017年10月

Tertiary lymphoid structures in ameloblastoma 査読

C. H. Siar, Z. A. Bin Abdul Rahman, H. Tsujigiwa, H. Nagatsuka, K. H. Ng

VIRCHOWS ARCHIV 471 S2 - S2 2017年9月

Semaphorin 4D promotes bone invasion in head and neck squamous cell carcinoma 国際誌

Hiroyuki Takada, Soichiro Ibaragi, Takanori Eguchi, Tatsuo Okui, Kyoichi Obata, Masanori Masui, Ayaka Morisawa, Kiyofumi Takabatake, Hotaka Kawai, Norie Yoshioka, Nur Mohammad Monsur Hassan, Tsuyoshi Shimo, Guo Fu Hu, Hitoshi Nagatsuka, Akira Sasaki

International Journal of Oncology 51 ( 2 ) 625 - 632 2017年8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：SPANDIDOS PUBL LTD

Head and neck squamous cell carcinomas (HNSCCs) frequently invade the bones of the facial skeleton. Semaphorin 4D (Sema4D) is an axon guidance molecule produced by oligodendrocytes. Sema4D was also identified in the bone microenvironment and many cancer tissues including HNSCC. To date, however, the role of Sema4D in cancer-associated bone disease is still unknown. This is the first study to demonstrate the role of Sema4D in bone invasion of cancer. In the clinical tissue samples of bone lesion of HNSCC, Sema4D was detected at high levels, and its expression was correlated with insulin-like growth factor-I (IGF-I) expression. In vitro experiments showed that IGF-I regulates Sema4D expression and Sema4D increased proliferation, migration and invasion in HNSCC cells. Sema4D also regulated the expression of receptor activator of nuclear factor κβ ligand (RANKL) in osteoblasts, and this stimulated osteoclastgenesis. Fur thermore, knockdown of Sema4D in HNSCC cells inhibited tumor growth and decreased the number of osteoclasts in a mouse xenograft model. Taken together, IGF-I-driven production of Sema4D in HNSCCs promotes osteoclastogenesis and bone invasion.

DOI： 10.3892/ijo.2017.4050

MCSF orchestrates branching morphogenesis in developing submandibular gland tissue 国際誌

Gulsan Ara Sathi, Mahmoud Farahat, Emilio Satoshi Hara, Hiroaki Taketa, Hitoshi Nagatsuka, Takuo Kuboki, Takuya Matsumoto

Journal of Cell Science 130 ( 9 ) 1559 - 1569 2017年5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：COMPANY OF BIOLOGISTS LTD

The importance of macrophages in tissue development and regeneration has been strongly emphasized. However, the specific roles of macrophage colony-stimulating factor (MCSF), the key regulator of macrophage differentiation, in glandular tissue development have been unexplored. Here, we disclose new macrophage-independent roles of MCSF in tissue development. We initially found that MCSF is markedly upregulated at embryonic day (E)13.5, at a stage preceding the colonization of macrophages (at E15.5), in mouse submandibular gland (SMG) tissue. Surprisingly, MCSF-induced branching morphogenesis was based on a direct effect on epithelial cells, as well as indirectly, by modulating the expression of major growth factors of SMG growth, FGF7 and FGF10, via the phosphoinositide 3-kinase (PI3K) pathway. Additionally, given the importance of neurons in SMG organogenesis, we found that MCSF-induced SMG growth was associated with regulation of neurturin expression and neuronal network development during early SMG development in an in vitro organogenesis model as well as in vivo. These results indicate that MCSF plays pleiotropic roles and is an important regulator of early SMG morphogenesis.

DOI： 10.1242/jcs.196907

De novo myoepithelial carcinoma with multiple metastases arising from a submandibular salivary gland: A case report 国際誌

Karina Cecília Panelli Santos, Hidenobu Matsuzaki, Teruhisa Unetsubo, Shimo Tsuyoshi, Hitoshi Nagatsuka, Jun Ichi Asaumi

Oncology Letters 13 ( 4 ) 2679 - 2683 2017年4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：SPANDIDOS PUBL LTD

Salivary gland carcinomas are rare tumors, representing ~0.5% of all malignancies. Myoepithelioma is also representing ~1% of all salivary gland tumors. Myoepithelial carcinoma (MC) is even rarer, representing 0.2 to 0.6% of all salivary gland tumors. We herein report a case of MC with multiple metastases arising from a submandibular gland in a 71-year-old male patient and present the associated imaging findings. The patient was considered to have a de novo type of myoepithelial carcinoma, which is reportedly associated with higher malignancy than the transformation type of the disease (i.e., a malignant change from pleomorphic adenoma or myoepithelioma). This was reflected in the multiple lung and bone metastases sites and strong positivity for p53 and Ki-67.

DOI： 10.3892/ol.2017.5783

miRNA-mediated crosstalk between Wnt3a and TGFβ3 in osteoblast differentiation

Fushimi S, Nohno T, Nishimatsu S, Katase N, Terada K, Katsuyama M, Demura M, Saijoh K, Nagatsuka H, Katsuyama H

The FASEB Journal 31 ( 1Suppl ) 757.3 2017年4月

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