Faculty Profiles - NAGATSUKA Hitoshi

Papers - NAGATSUKA Hitoshi

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Histological evaluation of periodontal ligament in response to orthodontic mechanical stress in mice Reviewed International journal

Keiko Kaneko, Saeka Matsuda, Rina Muraoka, Keisuke Nakano, Takami Iwasaki, Mihoko Tomida, Hidetsugu Tsujigiwa, Hitoshi Nagatsuka, Toshiyuki Kawakami

International Journal of Medical Sciences 12 ( 9 ) 689 - 694 2015.8

Language：English Publishing type：Research paper (scientific journal)

The purpose of the study was to determine the cell dynamics in periodontal ligament in response to mechanical stress during orthodontic movement. Following Waldo’s method, a square sheet of rubber dam was inserted in between the first and second maxillary molars in 10 ddY mice leaving the stress load for 3 hours. After 3 days and at 1 week, cell count on pressure and tension sides of the periodontal ligament was determined. Furthermore, the type of cell present after mechanical stress was identified using GFP bone marrow transplantation mouse model. Immunohistochemistry was carried out at 0 min (immediately after mechanical stress), 24 hours, 1 week, 2 weeks and 6 months. Temporal changes in the expression of GFP-positive bone marrow derived cells were examined. Moreover, double immunofluorescent staining was performed to determine the type of cell in the periodontal ligament. Cell count on the tension side tremendously increased 3 days after mechanical stress. At 1 week, spindle and round cell count increased compared to the control group. These changes were observed on both tension and pressure sides. Cell count on pressure side at 3 days (22.11+/-13.98) and at 1 week (33.23+/-11.39) was higher compared to the control group (15.26+/-8.29). On the tension side, there was a significantly increased at 3 days (35.46+/-11.85), but decreased at 1 week (29.23+/-13.89) although it is still higher compared to the control group (AD+/-SD: 10.37+/-8.69). Using GFP bone marrow transplantation mouse model, GFP positive cell count increased gradually over time in 6 months. GFP positive cells were also positive to CD31, CD68 and Runx2 suggesting that fibroblasts differentiated into osteoclasts and tissue macrophages. In conclusion, mechanical stress during orthodontic movement promoted the increase in the number of cells in the periodontal ligament on both tension and pressure sides. The increase in the number of cells in the periodontal ligament is believed to be due to the migration and cell division of undifferentiated mesenchymal cells.

DOI： 10.7150/ijms.12883

Early Initiation of Endochondral Ossification of Mouse Femur Cultured in Hydrogel with Different Mechanical Stiffness International journal

Gulsan Ara Sathi, Kodai Kenmizaki, Satoshi Yamaguchi, Hitoshi Nagatsuka, Yasuhiro Yoshida, Aira Matsugaki, Takuya Ishimoto, Satoshi Imazato, Takayoshi Nakano, Takuya Matsumoto

Tissue Engineering - Part C: Methods 21 ( 6 ) 567 - 575 2015.6

Language：English Publishing type：Research paper (scientific journal) Publisher：MARY ANN LIEBERT, INC

Mineralization is one of the most important processes in normal bone tissue development and in disease condition. Developing a novel and standardized in vitro model system that can readily monitor both cellular dynamics and mineralization is crucial for better understanding the bone tissue development and growth. Recent studies indicated that the mechanical environment is a critical condition in mineralization. We hypothesized that hydrogel with different mechanical stiffness can provide a biomimetic mechanical environment that can modulate bone tissue growth and mineralization. A femur of mouse embryo (embryonic day 16) was embedded in agarose hydrogel (2-60 kPa) and cultured in an osteogenic medium for a week. Microcomputed tomography (μCT) results revealed enhanced mineralization was detected in the femur head cultured in the gel condition, whereas no mineralization in the femur head cultured in the control (floating culture) condition. The mineralized region was corresponding to the region of secondary ossification center. Both histological and quantitative analyses indicated that the mineralized region of femur head cultured in 10 kPa gel condition was the highest and the mineralized area was significantly larger than that cultured in 2, 40, and 60 kPa gel condition. Immunofluorescence results indicated the enhanced mineralization caused by the higher chondrogenic differentiation at that region. This enhancement mainly relating to the mechanical forces and not to the oxygen tension was also confirmed. Since this system enhances and shortens the mineralization procedure compared with the conventional two-dimensional or three-dimensional cell culture system, this hydrogel system would be one of the unique models for better understanding the mineralized tissue development.

DOI： 10.1089/ten.tec.2014.0475

Long-term bioresorption of bone fixation devices made from composites of unsintered hydroxyapatite particles and poly-L-lactide Reviewed

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

Journal of Hard Tissue Biology 24 ( 2 ) 219 - 224 2015.4

Language：English Publishing type：Research paper (scientific journal) Publisher：JOURNAL HARD TISSUE BIOLOGY

Osteosynthetic bone fixation devices made from composites of uncalcined and unsintered hydroxyapatite (u-HA) particles and poly-L-lactide (PLLA) are widely adopted for clinical use because of their bioresorbability and osteoconductive properties. However, how the plate systems constituting these devices change during long-term use in vivo is unknown. In this study, we present cases of two patients fitted with u- HA/PLLA devices for >5 years after surgery and evaluate the resorption process on the basis of the residual versus the resorbed material. In both cases, the majority of the degraded plates and screws had been replaced by bone. In post-operative three-dimensional (3D) CT imaging, plate and screws were maintained until two years after surgery, and then they were degraded and replaced to bone in 4-6 years after surgery. Examination of the aggregation of hydroxyapatite and decrease in molecular weight suggested that the residual material was in the final stages of resorption. The plate system examined demonstrated stable degradation without foreign body reactions in vivo. Although complete resorption is a lengthy process, it is possible to follow its progress using CT.

DOI： 10.2485/jhtb.24.219

RANK, RANKL, and OPG in recurrent solid/multicystic ameloblastoma: Their distribution patterns and biologic significance International journal

Chong Huat Siar, Hidetsugu Tsujigiwa, Ismadi Ishak, Nurmawarnis Mat Hussin, Hitoshi Nagatsuka, Kok Han Ng

Oral Surgery, Oral Medicine, Oral Pathology and Oral Radiology 119 ( 1 ) 83 - 91 2015.1

Language：English Publishing type：Research paper (scientific journal) Publisher：ELSEVIER SCIENCE INC

Objectives To determine the distribution patterns of bone resorption regulators, receptor activator of nuclear factor κ-B (RANK), RANK ligand (RANKL), and osteoprotegerin (OPG) in recurrent ameloblastoma (RAs) and to clarify their impact on the biologic behavior of these neoplasms.

DOI： 10.1016/j.oooo.2014.09.017

Effect of sintering time of CaTiO<inf>3</inf>–CaCO<inf>3</inf> in osteoblastic response of KUSA/A1 cells Reviewed

Andrea P. Rodríguez, María A. Sánchez, Miho Inoue, Betiana Felice, Hitoshi Nagatsuka, Rossana E. Madrid, Carmelo Felice, Hidetsugu Tsujigiwa

IFMBE Proceedings 49 159 - 162 2015

Language：English Publishing type：Research paper (international conference proceedings) Publisher：SPRINGER INT PUBLISHING AG

In recent years, calcium titanate (CaTiO3) and carbon-containing materials have gained much attention in a number of biomedical material researches. To maximize the advantages of both materials, we developed a novel alkoxide method to get ‘‘calcium titanate with calcium carbonate’’ (CaTiO3–CaCO3). Thus, our previous result indicated that calcium carbonate could play a key role in the cell response of CaTiO3 material. In the present study we evaluated the effect of sintering time of CaTiO3–CaCO3 in osteoblastic response of KUSA/A1 cells. We examined the material characterization as well as cellular proliferation and mineralization of KUSA/A1 cells cultured with the materials. The results showed that Ca-TiO3–CaCO3 material sintered during 30 min, has better influ-ence on the bone marrow stromal cells for their proliferation and mineralization compared to CaTiO3-CaCO3 sintered at 2 and 10 h. These results suggest that sintering time could be directly related to the amount of carbon within CaCO3 being crucial in cell response. In conclusion, our preliminary find-ings indicate that CaTiO3–CaCO3 could play a more dominant and favorable effect on cell response when sintered for 30 minutes.

DOI： 10.1007/978-3-319-13117-7_42

A case of adenoid cystic carcinoma associated with IgG4-related disease Reviewed International journal

Tsuyoshi Shimo, Mayumi Yao, Yuichiro Takebe, Yuko Ono, Kyoichi Obata, Naito Kurio, Soichiro Ibaragi, Norie Yoshioka, Koji Kishimoto, Yoshinobu Yanagi, Hitoshi Nagatsuka, Akira Sasaki

International Journal of Surgery Case Reports 10 12 - 16 2015

Language：English Publishing type：Research paper (scientific journal) Publisher：ELSEVIER SCI LTD

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.

DOI： 10.1016/j.ijscr.2015.01.022

Does HMGB1 predict occult neck lymph node metastasis in early tongue carcinoma? A case-control study of 26 patients

H. Hanakawa, Y. Orita, Y. Sato, M. Takeuchi, S. Takao, K. Ohno, T. Kohno, N. Iwaki, H. Marunaka, R. Tamamura, M. Nishibori, H. Nagatsuka, K. Nishizaki, T. Yoshino

Journal of Laryngology and Otology 128 ( 10 ) 926 - 931 2014.10

Publishing type：Research paper (scientific journal)

Objective: This study examined whether the occurrence of late neck metastasis in early tongue squamous cell carcinoma can be predicted by evaluating HMGB1 (high mobility group box 1) expression in the primary lesion. Methods: A case-control study was conducted. The cases comprised 10 patients with late neck metastasis. The controls consisted of 16 patients without recurrence. All were examined immunohistochemically for HMGB1 protein expression. The odds ratio for late neck metastasis in relation to HMGB1 was estimated. Results: Results for HMGB1 were dichotomised into positive staining scores (score, 5-7) and negative scores (0-4). Six cases (60 per cent) and four controls (25 per cent) were HMGB1-positive. Although no significant result was seen, compared with HMGB1-negative patients the odds ratio for late neck metastasis in HMGB1-positive patients was 3.8 (95 per cent confidence interval, 0.6-26.5) after adjusting for other factors. Conclusion: In the present study, immunohistochemical study of HMGB1 in early tongue squamous cell carcinoma did not appear to be very useful for predicting occult neck metastasis. Further study is necessary to clarify the relationship between HMGB1 expression and late neck metastasis in early tongue squamous cell carcinoma.

DOI： 10.1017/S0022215114001819

DKK3 expression and its function in head and neck squamous cell carcinoma Reviewed

Naoki Katase, Mehmet Gunduz, Hidetsugu Tsujigiwa, Satoshi Ito, Masae Fujii, Hitoshi Nagatsuka, Akira Sasaki, Tsutomu Nohno

CANCER RESEARCH 74 ( 19 ) 2014.10

移植骨髄由来細胞の歯周組織への遊走と細胞分化(Transplanted bone marrow-derived cell migration into periodontal tissue and cell differentiation)

Tsujigiwa Hidetsugu, Muraoka Rina, Nakano Keisuke, Tomida Mihoko, Takabatake Kiyofumi, Tamamura Ryo, Nagatsuka Hitoshi, Kawakami Toshiyuki

Journal of Hard Tissue Biology 23 ( 4 ) 470 - 470 2014.10

エナメル上皮腫細胞による腫瘍間質の多様性の解析(Analyze of variation of the tumor stroma by ameloblastoma cells)

Takebe Yuichiro, Kawai Hotaka, Takabatake Kiyofumi, Song Yu, Ito Satoshi, Huji Masae, Tsujigiwa Hidetsugu, Nagatsuka Hitoshi

Journal of Hard Tissue Biology 23 ( 4 ) 470 - 471 2014.10

Effect of geometry and microstructure of honeycomb TCP scaffolds on bone regeneration International journal

Kiyofumi Takabatake, Eiki Yamachika, Hidetsugu Tsujigiwa, Yasushi Takeda, Mariko Kimura, Shin Takagi, Hitoshi Nagatsuka, Seiji Iida

Journal of Biomedical Materials Research - Part A 102 ( 9 ) 2952 - 2960 2014.9

Language：English Publishing type：Research paper (scientific journal) Publisher：WILEY

In recent years, artificial biological materials have been commonly used for the treatment of bone tissue defects caused by trauma, tumors, or surgical stress. Although tricalcium phosphate (TCP) is a promising absorbent bone tissue reconstruction biomaterial, it has been reported that its biocompatibility and osteoconductivity depend on its preparation method and sintering temperature. In addition, although it is thought that the microenvironment produced by the extracellular matrix plays an important role in cell growth and differentiation, there have been few studies on how the geometric structure of artificial biological materials affects cells. In the present study, a new honeycomb TCP scaffold containing through-holes with diameters of 300 μm has been developed. The influence of the sintering temperature on the crystal structure and material properties of the honeycomb TCP scaffold was investigated using scanning electron microscopy and X-ray diffraction. Its biocompatibility and osteoconductivity were also evaluated by implantation into experimental animals. It was found that a β-TCP scaffold sintered at 1200°C exhibited high biocompatibility and osteoconductivity, and when it was loaded with BMP-2, it exhibited both osteoconductivity and osteoinductivity, promoting rapid bone formation in both ectopic and orthotopic areas. It is thus a highly promising bone reconstruction material that is expected to find clinical applications. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 2952-2960, 2014. © 2013 Wiley Periodicals, Inc.

DOI： 10.1002/jbm.a.34966

Regulatory T-cell infiltration in tongue squamous cell carcinoma Reviewed International journal

Hiroyuki Hanakawa, Yorihisa Orita, Yasuharu Sato, Mai Takeuchi, Kyotaro Ohno, Yuka Gion, Kiyoaki Tsukahara, Ryo Tamamura, Toshihiro Ito, Hitoshi Nagatsuka, Kazunori Nishizaki, Tadashi Yoshino

Acta Oto-Laryngologica 134 ( 8 ) 859 - 864 2014.8

Language：English Publishing type：Research paper (scientific journal)

Conclusion: In tongue squamous cell carcinoma (SCC), high levels of regulatory T-cell (Treg) infiltration in tumor nests are observed in the cases with poor prognosis. Objectives: The role of Tregs in head and neck cancers remains unclear. The aim of this study was to observe the distribution of Tregs in different stages of tongue SCC and estimate the effects on prognosis. Methods: Thirty-four cases with tongue SCC were examined immunohistochemically for CD4, CD8, and Forkhead box P3 (Foxp3). Immunoreactive cells were counted in cancer stroma and nest regions, and relationships between cell numbers and disease-free survival rates were analyzed. Results: In the 34 cases, univariate analysis for disease-free survival indicated high-level infiltration of Tregs (CD4+Foxp3+) into both cancer nests and stroma and presence of helper T (CD4+Foxp3-) cells in cancer stroma as potential predictors of significantly worse prognosis. In early-stage cases (stage I/II), high-level infiltration of Tregs in cancer nests correlated significantly with poor disease-free survival rate. Multivariate analysis for disease-free survival found no independent variables. © Informa Healthcare.

DOI： 10.3109/00016489.2014.918279

口腔癌細胞株担癌マウスに対するBCG生菌と5-FU併用療法による延命効果の検討

村上純, 大原直也, 中山真彰, 辻極秀次, 長塚仁, 此内浩信, 柳文修, 苔口進, 久富美紀, 藤田麻里子, 畦坪輝寿, 浅海淳一

日本癌治療学会誌 49 ( 3 ) 1533 - 1533 2014.6

Locally injected dexmedetomidine inhibits carrageenin-induced inflammatory responses in the injected region International journal

Shintaro Sukegawa, Hitoshi Higuchi, Miho Inoue, Hitoshi Nagatsuka, Shigeru Maeda, Takuya Miyawaki

Anesthesia and Analgesia 118 ( 2 ) 473 - 480 2014.2

Language：English Publishing type：Research paper (scientific journal) Publisher：LIPPINCOTT WILLIAMS & WILKINS

BACKGROUND:: Dexmedetomidine, a highly selective agonist of α2-adrenoceptors, is a commonly used sedative; however, a potent anti-inflammatory effect has also been found. In the present study we evaluated the inhibitory effect of locally injected dexmedetomidine on inflammatory responses in the injected region. METHODS:: Local inflammation was induced in the hindpaws of male mice (aged 6-8 weeks) by intraplantar injection of lambda-carrageenin. To offset the central effect of tested agents, different agents were blindly injected into the left and right paws in the pairs of comparison. The effect of dexmedetomidine on edema (increase in paw volume), the accumulation of leukocytes, and production of tumor necrosis factor-α (TNF-α) and cyclooxygenase-2 (COX-2) were evaluated after carrageenin injection, using water displacement plethysmometry, histological imaging, immunohistochemistry, and Western blotting analysis. Furthermore, we also evaluated the effect of yohimbine, a full antagonist of α2-adrenoceptors, and phenylephrine, an agonist of the α1-adrenoceptor, on dexmedetomidine's action on inflammatory responses. RESULTS:: Paw volume and amount of leukocytes in the injected region significantly increased after the injection of carrageenin. Similarly, TNF-α and COX-2 production was found in the subcutaneous region injected with carrageenin, 4 hours after injection. Dexmedetomidine significantly inhibited all increases in paw volume, leukocytes, and production of TNF-α and COX-2. Furthermore, yohimbine significantly antagonized the anti-inflammatory effects of dexmedetomidine, whereas phenylephrine did not significantly alter them. CONCLUSIONS:: The findings suggest that locally injected dexmedetomidine exhibits an anti-inflammatory effect against local acute inflammatory responses, mediated by α2-adrenoceptors. © 2013 International Anesthesia Research Society.

DOI： 10.1213/ANE.0000000000000060

Expression and role of sonic hedgehog in oral squamous cell carcinoma induced jaw bone destruction. Reviewed

Tsuyoshi Shimo, Naito Kurio, Masahiro Iwamoto, Tatsuo Okui, Hiromasa Kuroda, Kenichi Matsumoto, Soichiro Ibaragi, Norie Yoshioka, Yuichirou Takebe, Hitoshi Nagatsuka, Akira Sasaki

JOURNAL OF BONE AND MINERAL RESEARCH 29 S372 - S372 2014.2

Expression and roles of CCN2 in dental mesenchymal cells in primary culture-With findings in a case of odontogenic myxofibroma Reviewed

Tsuyoshi Shimo, Eiki Koyama, Yuu Horikiri, Tatsuo Okui, Naito Kurio, Naoki Katase, Shoko Yoshida, Yuichiro Takebe, Koji Kishimoto, Norie Yoshioka, Hitoshi Nagatsuka, Akira Sasaki

Oral Science International 11 ( 1 ) 8 - 14 2014.1

Language：English Publishing type：Research paper (scientific journal)

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. © 2013 Japanese Stomatological Society.

DOI： 10.1016/S1348-8643(13)00008-6

Fibrous dysplasia of the maxilla: A case report together with its conventional imaging and dynamic magnetic resonance imaging findings Reviewed

Marina Hara, Hidenobu Matsuzaki, Naoki Katase, Teruhisa Unetsubo, Yoshinobu Yanagi, Hitoshi Nagatsuka, Jun Ichi Asaumi

Oral Radiology 30 ( 1 ) 105 - 110 2014.1

Language：English Publishing type：Research paper (scientific journal)

We previously reported that dynamic contrastenhanced MRI parameters and time-signal intensity curves (TICs; also known as contrast index curves) are useful for the differential diagnosis of jawbone lesions. In particular, odontogenic fibroma and ossifying fibroma, which possess similar histopathological features (i.e., a mixture of hard and soft tissue components), display unique TIC patterns, and we consider that the TIC patterns of these lesions reflect their hard and soft tissue components. Therefore, fibrous dysplasia, which contains fibrous tissue and immature isolated trabeculae composed of woven bone, is expected to display an interesting TIC. The purpose of this study was to assess the utility of TICs for differentiating between the abovementioned lesions, which have similar histopathological components. © Japanese Society for Oral and Maxillofacial Radiology and Springer Japan 2013.

DOI： 10.1007/s11282-013-0137-1

Freeze dried bone matrix on rat critical size defect regeneration

María V. Jammal, Liliana R. Missana, Kiyofumi Takabatake, Shin Takagi, Hitoshi Nagatsuka

Journal of Hard Tissue Biology 23 ( 2 ) 233 - 237 2014

Publishing type：Research paper (scientific journal)

Bone allografts are commonly used for bone regeneration. The aim of this study was evaluate the efficacy of a freeze dried bone matrix (FDBM) in critical size defect (CZD) rat calvaria. Eighteen Wistar female rats (body weight 150 ± 50 g) with CZD (5mm) were divided in two groups: group 1, using freeze dried bone matrix; and group 2, only with coagulum. All samples were evaluated on the 1st, 3rd and 6th weeks post-surgery by soft X-ray, histological and histometric studies. Soft X-ray results showed a radiolucent image with many irregular radiopaque areas. Histologically, bone regeneration was initiated from the 3rd week, when a thin layer of new woven bone could be seen adjacent to the matrix. At the 6th week, lamellar bone covered over half (61.8%) of the defect area. The lack of FDBM resorption allowed its incorporation to the new regenerated bone. This behavior is important in circumstances where it is necessary not only to stimulate bone regeneration but also increase the volume in affected areas, such as during the placement of dental implants. The results obtained in this research are encouraging for the use of freeze dried bone matrix as a bone graft material. © 2014 The Hard Tissue Biology Network Association Printed in Japan, All rights reserved.

DOI： 10.2485/jhtb.23.233

Expressions of ABCG2, CD133, and podoplanin in salivary adenoid cystic carcinoma Reviewed International journal

Wuwei Li, Ryo Tamamura, Bo Wang, Qigui Liu, Han Liu, Tingjiao Liu, Naoki Katase, Jing Xiao, Hitoshi Nagatsuka

BioMed Research International 2014 132349 - 132349 2014

Language：English Publishing type：Research paper (scientific journal)

Adenoid cystic carcinoma (ACC) is one of the most common salivary gland malignant tumors with a high risk of recurrence and metastasis. Current studies on cancer stem cells (CSCs) have verified that CSCs are the driving force behind tumor initiation and progression, suggesting that new cancer therapies may be established by effectively targeting and killing the CSCs. The primary goal of this study is to investigate the expression patterns of ABCG2, CD133, and podoplanin in ACC of minor salivary glands by immunohistochemistry analysis. We found that ABCG2 was weakly expressed in normal looking salivary gland tissues. A significant upregulation of ABCG2 expression in ACC was observed with a similar expression pattern of Ki-67. CD133 was detected in apical membrane of epithelial cells and podoplanin was expressed positively in myoepithelial cells of both normal looking tissue and ACC. However, no significant difference was found of the expression pattern of CD133 and podoplanin between normal looking tissues and ACC. Our observations suggest that CSCs may exist in quiescent cells with ABCG2 positive staining, which are surrounded by cells with positive expression of ABCG2 and Ki-67 in ACC, and costaining with ABCG2 and Ki-67 may help predict the location of CSCs. © 2014 Wuwei Li et al.

DOI： 10.1155/2014/132349

Establishment of odontoblastic cells, which indicate odontoblast features both in vivo and in vitro International journal

Hidetsugu Tsujigiwa, Naoki Katase, Mathieu Lefeuvre, Eiki Yamachika, Ryo Tamamura, Satoshi Ito, Yuichiro Takebe, Hiroyuki Matsuda, Hitoshi Nagatsuka

Journal of Oral Pathology and Medicine 42 ( 10 ) 799 - 806 2013.11

Language：English Publishing type：Research paper (scientific journal) Publisher：WILEY-BLACKWELL

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. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

DOI： 10.1111/jop.12080

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