Faculty Profiles - NAGATSUKA Hitoshi

Papers - NAGATSUKA Hitoshi

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Gene expression of type 1 collagen in neoplastic chondrocytes.

T. Kawakami, A. Kimura, H. Hasegawa, H. Tsujigiwa, H. Nagatsuka, N. Nagai

JOURNAL OF DENTAL RESEARCH 82 B298 - B298 2003.6

Heparanase expression correlates with invasion and poor prognosis in gastric cancers

Munenori Takaoka, Yoshio Naomoto, Takaomi Ohkawa, Hirokazu Uetsuka, Yasuhiro Shirakawa, Futoshi Uno, Toshiyoshi Fujiwara, Mehmet Gunduz, Hitoshi Nagatsuka, Motowo Nakajima, Noriaki Tanaka, Minoru Haisa

Laboratory Investigation 83 ( 5 ) 613 - 622 2003.5

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

Degradation of basement membrane and extracellular matrix structures are important features of the metastatic process of malignant tumors. Human heparanase degrades heparan sulfate proteoglycans, which represent the main components of basement membranes and the extracellular matrix. Because of the role of heparanase in tumor invasion and metastasis, we examined heparanase expression in primary gastric cancers and in cell lines derived from gastric cancers by immunohistochemistry and RT-PCR, respectively. Four of seven gastric cancer cell lines showed heparanase mRNA expression by RT-PCR. Heparanase protein was detected in both the cytoplasm and the nucleus of heparanase mRNA-positive cells by immunohistochemical staining. Heparanase expression was confirmed in 35 (79.5%) of 44 gastric tumor samples by immunohistochemical staining. However, no or weak heparanase expression was detected in normal gastric mucosa. In situ hybridization showed that the mRNA expression pattern of heparanase was similar to that of the protein, suggesting that increased expression of the heparanase protein at the invasive front was caused by an increase of heparanase mRNA in tumor cells. Analysis of the clinicopathologic features showed stronger heparanase expression in cases of huge growing tumors, extensive invasion to lymph vessels, and regional lymph node metastasis. In gastric cancer, patients with heparanase expression showed significantly poorer prognosis than those without such expression (p = 0.006). In conclusion, our findings suggest that high expression of heparanase in gastric cancer is a strong predictor of poor survival.

DOI： 10.1097/01.LAB.0000067482.84946.BD

花北朋哉, 岡藤正樹, 内田堅一郎, 伊田正道, 上山吉哉, 早津良和, 長塚仁

日本口腔科学会雑誌 52 ( 3 ) 160 - 160 2003.5

Gene expression of type I collagen in neoplastic chondrocytes.

A. Kimura, Toshiyuki Kawakami, S. Matsuura, H. Hasegawa, H. Kanda, H. Tujigiwa, H. Nagatsuka, N. Nagai

European journal of medical research 8 ( 4 ) 165 - 167 2003.4

Language：English Publishing type：Research paper (scientific journal) Publisher：I HOLZAPFEL VERLAG GMBH

In the present examination, we detected type I collagen mRNA in neoplastic chondrocytes in osteochondromas, typical benign bone neoplasms. We believe that the cells involved in >chondroid bone< appearing in osteochondromas temporally express cartilage phenotypes and then change directly into bone-forming cells that survive in the >chondroid bone< until the tissue is resorbed and remodelled into true bone tissue.

Mastoid obliteration by BMP-2/collagen composites: An experimental study using tissue engineering

Kazunori Nishizaki, Hidetsugu Tsujigiwa, Yasushi Takeda, Tadashi Yoshino, Manabu Maeta, Kunihiro Fukushima, Hitoshi Nagatsuka, Noriyuki Nagai

American Journal of Otolaryngology - Head and Neck Medicine and Surgery 24 ( 1 ) 14 - 18 2003

Language：English Publishing type：Research paper (scientific journal) Publisher：W B SAUNDERS CO

Purpose: Several materials have been used in the application of mastoid cavity obliteration during surgery for cholesteatoma; however, nothing has won universal acceptance. Through the advancement of tissue engineering, bone morphogenetic protein-2 (BMP-2)/collagen composites have been elucidated as inducers of heterogenic bone formation. This study was performed to investigate whether these composites are potentially obliteration materials for use in the mastoid cavity by using an animal experimental study. Materials and Methods: The composites were implanted in the rat mastoid to investigate whether new bone would be tissue engineered in the mastoid and, if so, whether the newly formed bone was stable. The composites were examined histologically over a 24-week period. Results: The composites implanted in the rat mastoid were able to tissue engineer new bone, and the newly formed bone was stable as assessed histologically, with almost normal bone structure, that was not resorbed during the 24-week period. Adverse immunological reactions were not found during our observation. Conclusions: Bone that was tissue engineered by the BMP-2/collagen composites was stable as assessed by histological examination and persisted in the rat mastoid. The present study shows that the composites have the potential to become real materials for use in mastoid obliteration.

DOI： 10.1053/ajot.2003.13

Presence of human β-defensin-2 in oral lichen planus and its histamine releasing effect

Nobuyoshi Mizukawa, Koichi Sawaki, Hitoshi Nagatsuka, Eiki Yamachika, Takaaki Ueno, Joji Fukunaga, Shin Takagi, Toshio Sugahara

Asian Journal of Oral and Maxillofacial Surgery 15 ( 2 ) 112 - 117 2003

Language：English Publishing type：Research paper (scientific journal) Publisher：Scientific Communications International Ltd

Objective: To determine by immunohistochemical means the localization of human β-defensin-2, a peptide with antimicrobial activity, in oral lichen planus. The release of histamine from mast cells elicited by human β-defensin-2 was also investigated. Materials and Methods: Biopsy specimens of oral lichen planus, synthetic human β-defensin-2, human α-defensin-1, and mast cells isolated from Sprague Dawley rats were used. Tissue sections were embedded in paraffin and immunostained by the streptavidin-biotin-coupled peroxidase method. Isolated rat mast cells were injected with synthetic human β-defensin-2 and human α-defensin-1. Evans blue dye was injected into the tail vein of Sprague Dawley rats, followed by various doses of histamine, human β-defensin-2, human αdefensin-1, and saline. Results: Epithelial cells in lichen planus from the corneal layer to the spinous layer, were positively stained with anti-human β-defensin-2 antibody. Mast cells in subepithelial areas were also stained by anti-human β-defensin-2 antibody. Human β-defensin-2-induced histamine release from the isolated rat mast cells occurs in a dose-dependent manner. When human β-defensin-2 was injected into rat skin intradermally, the vascular permeability increased. These responses were completely abolished upon injection of the antihistamine drug diphenhydramine hydrochloride. Conclusion: Human β-defensin-2 and histamine may play an important role in the formation of oral lichen planus.

DOI： 10.1016/S0915-6992(03)80019-5

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Angioarchitecture of primary oral malignant melanomas International journal

You Jin Lee, Noriyuki Nagai, Chong Huat Siar, Keisuke Nakano, Hitoshi Nagatsuka, Hidetsugu Tsujigiwa, Cheng Hsiung Roan, Mehmet Gunduz

Journal of Histochemistry and Cytochemistry 50 ( 11 ) 1555 - 1562 2002.11

Language：English Publishing type：Research paper (scientific journal) Publisher：HISTOCHEMICAL SOC INC

Angiogenesis is an essential process in the progression of malignant tumors. However, little is known of the angioarchitecture in primary oral malignant melanoma. We sought to determine this by the use of periodic acid-Schiff (PAS) stain, endothelial markers (CD34, CD105) and laminin, and by transmission electron microscopy in two cases. The results demonstrated that endothelium-lined vessels dominated the tumor microvasculature and these stained positively for PAS, laminin, and endothelial markers. Mosaic and tumor-lined vessels were infrequently encountered. Most PAS-positive patterned networks and loops ultrastructurally represented intratumor microhemorrhages that probably arose secondary to tumor vessel leakiness. Vascular channels of the vasculogenic mimicry type were rare. They stained for laminin but not for endothelial markers.

DOI： 10.1177/002215540205001116

Elemental sulphur and alkali elutable melanin detected in oral melanosis and malignant melanoma by energy-filtering transmission electron microscopy

Noriyuki Nagai, You Jin Lee, Noriyuki Nagaoka, Mehmet Gunduz, Keisuke Nakano, Tetsundo Nojima, Hidetsugu Tsujigiwa, Esra Gunduz, Chong Huat Siar, Hitoshi Nagatsuka

Journal of Oral Pathology and Medicine 31 ( 8 ) 481 - 487 2002.9

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

Background: The morphology and contents of melanosomes are important features for differentiating melanocyte-derived melanotic lesions such as melanosis and malignant melanoma. Methods: In this study, we attempted to elucidate the structure of melanin and sulphur content in oral melanosis and malignant melanomas by ultrastructural analysis. Results: In oral melanosis, the essential pathological findings were overproduction of eumelanin and discharge of melanin into keratinocytes. In malignant melanoma in situ, pleomorphic and ellipsoid abnormal melanosomes with an increase in sulphur content and alkali elution rate were detected. In invasive malignant melanoma, the irregular ellipsoid and spheroid melanosomes existing either as discrete bodies or compound melanosomes with furtherly increased sulphur content and alkali elution were detected. Conclusions: Our findings suggest that abnormal melanosome morphology and high sulphur content are predictive markers for assessment of early or precancerous melanotic lesions and malignant melanoma.

DOI： 10.1034/j.1600-0714.2002.00010.x

Distribution of basement membrane type IV collagen α chains in ameloblastoma: An immunofluorescence study

Keisuke Nakano, Chong Huat Siar, Noriyuki Nagai, Ichiro Naito, Yoshikazu Sado, Hitoshi Nagatsuka, Choufuku Hoh, Katsuhiro Kurada, Hidetsugu Tsujigiwa, Mehmet Gunduz

Journal of Oral Pathology and Medicine 31 ( 8 ) 494 - 499 2002.9

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

Background: Type IV collagen, a heterotrimeric molecule that exists in six genetically distinct forms, α1(IV)-α6(IV) is a major structural component of basement membrane (BM) and acts as a scaffold for other BM constituents. Methods: Indirect immunofluorescence using α chain-specific monoclonal antibodies was employed to clarify basement membrane (BM) collagen IV distribution in two ameloblastoma, and for comparison, on oral mucosa and tooth germ. Results: Ameloblastoma BM expressed five of six genetically distinct forms of collagen IV: α1(IV), α2(IV), α5(IV) and α6(IV) chains occurred as intense linear stainings without disruption around neoplastic epithelium, and this expression pattern was fundamentally similar to oral mucosa BM; α4(IV) expression was rare and occurred around nests of primitive tumor cells or potentially invasive sites. The tooth germ demonstrated a stage- and position-specific collagen IV distribution: the inner enamel epithelium BM expressed α1(IV), α2(IV), and α4(IV) except in the cuspal predentine region; and the outer enamel epithelium BM expressed α1(IV), α2(IV), α5(IV), and α6(IV) chains. Conclusions: Results suggest that collagen IV α chain distribution in ameloblastoma BM plays an important role in tumor cytodifferentiation and progression.

DOI： 10.1034/j.1600-0714.2002.00162.x

The expression of dentin sialophosphoprotein gene in bone

C. Qin, J. C. Brunn, E. Cadena, A. Ridall, H. Tsujigiwa, H. Nagatsuka, N. Nagai, W. T. Butler

Journal of Dental Research 81 ( 6 ) 392 - 394 2002

Language：English Publishing type：Research paper (scientific journal) Publisher：SAGE PUBLICATIONS INC

Dentin sialoprotein (DSP) and dentin phosphoprotein (DPP) are expressed as a single mRNA transcript coding for a large precursor protein termed dentin sialophosphoprotein (DSPP). DSP, DPP, and DSPP have been considered to be tooth-specific. To test for the expression of the dspp gene in bone, we performed Western immunoblots and reverse-transcription polymerase chain-reaction (RT-PCR). With Western immunoblots, we detected DSP in the Gdm/EDTA extracts of rat long bone, at a level of about 1/400 of that in dentin. Using RT-PCR, we detected DSPP mRNA in mouse calvaria. Similar to Western immunoblots, the results of RT-PCR indicated that the dspp gene is expressed at a lower level in bone than in dentin and odontoblasts. Analysis of the data shows that DSPP is not a tooth-specific protein, and that dramatically different regulatory mechanisms governing DSPP expression are involved in the bone and dentin.

DOI： 10.1177/154405910208100607

Usefulness as guided bone regeneration membrane of the alginate membrane International journal

Yoshiya Ueyama, Kunio Ishikawa, Takamitsu Mano, Takahiro Koyama, Hitoshi Nagatsuka, Kazuomi Suzuki, Kazuo Ryoke

Biomaterials 23 ( 9 ) 2027 - 2033 2002

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

Alginate membrane is a new bioabsorbable, guided bone regeneration (GBR) membrane, which is placed directly on the surface of the bone defect. It is designed to drop a calcium chloride aqueous solution into the bone defect, which is filled with sodium alginate aqueous solution. Alginate membrane is an excellent agent for this procedure due to its close assimilation to the surface of the bone. In this study, we evaluated the short-term biocompatibility of alginate membrane in the bone defects of rat tibiae. GBR membrane availability was also examined. Consequently, we found that the healing process in bone defects covered with an alginate membrane was delayed in comparison with that of controls; however, the defect was restored to nearly original condition. In contrast, in the controls, bone defect repairs exhibited partitioning as a result of connective tissue involvement. Furthermore, we observed a relation between the sodium alginate concentration and the rate of absorption of the sodium alginate membrane. Absorption of a 1.5% sodium alginate membrane was slow. As a result, the compound was not absorbed completely and bone repairs resembled an hourglass. Moreover, the inflammatory response was absent surrounding the alginate membrane. The present findings suggested that the alginate membrane functions effectively as a GBR membrane. In addition, the alginate membrane derived from 3% calcium chloride and 1% sodium alginate was most suitable as a GBR membrane. © 2002 Elsevier Science Ltd. All rights reserved.

DOI： 10.1016/S0142-9612(01)00332-5

Immunohistochemical studies of a case of clear cell chondrosarcoma

Lamaroon A, Kumchai T, Prapayasatok S, Siripalangkanont S, Nagatsuka H, Nagai N

J. Hard Tissue Biology 2002

Differential expression of collagen IV α1 to α6 chains in basement membranes of benign and malignant odontogenic tumors

Hitoshi Nagatsuka, Chong Huat Siar, Keisuke Nakano, Hidetsugu Tsujigiwa, Mehmet Gunduz, Hoh Choufuku, You Jin Lee, Ichiro Naito, Yoshikazu Sado, Noriyuki Nagai

Virchows Archiv 441 ( 4 ) 392 - 399 2002

Language：English Publishing type：Research paper (scientific journal) Publisher：SPRINGER-VERLAG

Type IV collagen, the major component of basement membrane (BM), demonstrates a stage- and position-specific distribution of its isoforms during tooth development. To determine its localization in BM of odontogenic neoplasms, immunohistochemistry using six anti-α(IV) chain-specific monoclonal antibodies was performed. Results disclosed that BM demonstrated an irregular α(IV) chain profile in malignant odontogenic tumors as compared to benign odontogenic neoplasms. No α3(IV) chains were detected. Expression of α1(IV)/α2(IV) and α5(IV)/α6(IV) chains was stronger in desmoplastic (n=3) than in ordinary (n=5) ameloblastomas. The adenomatoid odontogenic tumor (n=2) distinctly expressed these chains in BM of cribriform areas and hyaline materials (which was also α4(IV)-positive), but weakly around epithelial whorls/rosettes/nests and mineralized foci. These five chains also stained BM and tumor cells of ameloblastic fibroma (n=3) and ameloblastic fibro-odontosarcoma (n=1), but not the inductive hard tissues. Ameloblastic carcinoma (n=2) showed specific α1(IV)/α2(IV) chain loss, while primary intra-osseous carcinoma (n=1) demonstrated a discontinuous α1(IV)/α2(IV) and α5(IV)/α6(IV) staining pattern. The present results suggest that modification and remodeling of BM collagen IVα chains occur during odontogenic neoplasms' progression.

DOI： 10.1007/s00428-002-0658-1

Initial tissue response to anti-washout apatite cement in the rat palatal region: Comparison with conventional apatite cement

Yoshiya Ueyama, Kunio Ishikawa, Takamitsu Mano, Takahiro Koyama, Hitoshi Nagatsuka, Tomohiro Matsumura, Kazuomi Suzuki

Journal of Biomedical Materials Research 55 ( 4 ) 652 - 660 2001.6

Language：English Publishing type：Research paper (scientific journal) Publisher：JOHN WILEY & SONS INC

Initial tissue response to anti-washout apatite cement (aw-AC) in the palatal region was studied. Conventional apatite cement (c-AC) was employed as a control material. Bone defects generated in the rat palatal region, where complete hemostasis is difficult to effect, were filled with both cement types and examined histologically for up to 8 weeks. At 1-week postfilling, a portion of the c-AC had washed out, resulting in slight inflammation and severe foreign-body response. The degree of foreign-body response to c-AC was reduced over time; however, foreign-body response continued to be in evidence 8 weeks after surgery. As a result, poor bone formation was observed in the case of c-AC at 8 weeks post-surgery. In contrast, aw-AC set well, maintained its shape at implantation, and caused little foreign-body response. Osteoblasts were observed at 2 weeks following surgery. Moreover, the bone defect was completely covered with new bone at 8 weeks post-surgery. This observation suggests that aw-AC may be used without complication in cases where complete hemostasis is difficult to achieve, that is, where the use of c-AC is contraindicated. © 2001 John Wiley & Sons, Inc.

DOI： 10.1002/1097-4636(20010615)55:4<652::AID-JBM1060>3.0.CO;2-K

Evidence for the expression of dentin sialophosphoprotein gene in osteoblasts

C Qin, E Cadena, A Ridall, Y Ishiwari, H Nagatsuka, N Nagai, WT Butler

BONE 28 ( 5 ) S147 - S147 2001.5

Expression of Type I, Type II, and Type X Collagen Genes during Altered Endochondral Ossification in the Femoral Epiphysis of Osteosclerotic (oc/oc) Mice

A. Yamasaki, M. Itabashi, Y. Sakai, H. Ito, Y. Ishiwari, H. Nagatsuka, N. Nagai

Calcified Tissue International 68 ( 1 ) 53 - 60 2001.1

Language：English Publishing type：Research paper (scientific journal) Publisher：SPRINGER-VERLAG

The osteosclerotic (oc/oc) mouse, a genetically distinct murine mutation that has a functional defect in its osteoclasts, also has rickets and shows an altered endochondral ossification in the epiphyseal growth plate. The disorder is morphologically characterized by an abnormal extension of hypertrophic cartilage at 10 days after birth, which is later (21 days after birth) incorporated into the metaphyseal woven bone without breakdown of the cartilage matrix following vascular invasion of chondrocyte lacunae. In situ hybridization revealed that the extending hypertrophic chondrocytes expressed type I and type II collagen mRNA, as well as that of type X collagen and that the osteoblasts in the metaphysis expressed type II and type X collagen mRNA, in addition to type I collagen mRNA. The topographic distribution of the signals suggests a possible co-expression of each collagen gene in the individual cells. Immunohistochemically, an overlapping deposition of type I, type II, and type X collagen was observed in both the extending cartilage and metaphyseal bony trabeculae. Such aberrant gene expression and synthesis of collagen indicate that pathologic ossification takes place in the epiphyseal/ metaphyseal junction of oc/oc mouse femur in different way than in normal endochondral ossification. This abnormality is probably not due to a developmental disorder in the epiphyseal plate but to the failure in conversion of cartilage into bone, since the epiphyseal plate otherwise appeared normal, showing orderly stratified zones with a proper expression of cartilage-specific genes.

DOI： 10.1007/BF02685003

Immunohistochemical Localization of Cytokeratin and PCNA in Desmoplastic Ameloblastoma.

Nagatsuka, H, Fujii, E, Tamamura, R, Hibi, K, Ishiwari, Y, Thunyakitpisal p, Sao S.G, Qi j.-p, Tancharoen S, Nagai, N

Journal of Dental Research 2001

Combination of tumor necrosis factor alpha and interferon alpha induces apoptotic cell death through a c-myc-dependent pathway in p53 mutant H226br non-small-cell lung cancer cell line

YASUOKA Y

Exp Cell Res 271 214 - 222 2001

Phenomelanin and Eumelanin of Malignant Melanomas and Melanomas in Oral Mucosa

You-Jin LEE, Noriyuki NAGAOKA, Toshiro SAKAE, Tohru TAKAGI, Hitoshi NAGATSUKA, Yasuo SHINNOU, Kazuteru HIBI, Noriyuki NAGAI, Department of oral Pathology Okayama University Dental School, Department of oral Pathology Okayama University Dental School, Department of Oral Anatomy Nihon University school of Dentistly at Matsudo, Department of oral Pathology Okayama University Dental School, Department of oral Pathology Okayama University Dental School, Department of oral Pathology Okayama University Dental School, Department of oral Pathology Okayama University Dental School

Journal of Hard Tissue Biology 10 ( 2 ) 89 - 95 2001

Characteristics of bone morphogenetic protein-induced chondroid bone: Histochemical, immunohistochemical and in situ hybridization examinations

T. Kawakami, T. Kawai, A. Kimura, H. Hasegawa, H. Tsujigiwa, M. Gunduz, H. Nagatsuka, N. Nagai

Journal of International Medical Research 29 ( 6 ) 480 - 487 2001

Language：English Publishing type：Research paper (scientific journal) Publisher：CAMBRIDGE MED PUBL

The third ossification mode, known as transchondroid bone formation, is displayed chiefly in the bone morphogenetic protein (BMP)-induced heterotopic bone formation model. This paper describes the results of histopathological, histochemical, immunohistochemical, and in situ hybridization examinations of BMP-induced heterotopic bone in mice. The research focuses on the localization of typical matrix proteins (peptide and its mRNA) of cartilage and bone - type-I and type-II collagen, osteocalcin and osteopontin - in the chondroid bone matrix.

DOI： 10.1177/147323000102900603

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Papers - NAGATSUKA Hitoshi