The 5th International Symposium on “Dental and
Craniofacial Morphogenesis and Tissue
Regeneration: A View from Stem Cell Research”

第５回国際シンポジウム「口腔組織の再生・再建医療研究」(文部科学省 特別教育研究経費「口腔から QOL の向上を目指す連携研究」)

February 5, 2010
Centennial Hall, Kyushu University School of
Medicine, Fukuoka, JapanPROGRAM & ABSTRACTS

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The 5th International Symposium on “Dental and
Craniofacial Morphogenesis and Tissue Regeneration:
A View from Stem Cell Research”■ 日時：平成２２年２月５日（金）9:30〜
18:30■ 場所：九州大学医学部

百年講堂

福岡市東区馬出 3-1-1（TEL: 092-642-6030）■ 主催：九州大学大学院

歯学研究院「口腔組織の再生・再建医療研究」プロジェクト
共催：文部科学省 特別教育研究経費「口腔から QOL の向上を目指す連携研究」
九州大学リサーチコア「口腔組織の再生医療の総合的開発」

■

プログラム

プロジェクト成果報告会（9:40〜11:40）
・ ランチョンセミナー（12:00〜13:00）共催：日機装株式会社
・Dr. Hideo Iwasaka（Oita University, Japan）

・特別講演（13:20〜18:10）
Dr. Yang Chai（University of Southern California, USA）
Dr. Richard O. Oreffo（University of Southampton, UK）
Dr. Songtao Shi (University of Southern California, USA)
Dr. Hidefumi Maeda (Kyushu University, Japan)
Dr. Takayoshi Yamaza (Kyushu University, Japan)

・ 交流懇親会（19:00〜20:30）
■

参加費無料
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Centennial Hall
Kyushu University
School of Medicine

九州大学医学部
百年講堂

Maidashi 3-1-1, Higashi-ku
Fukuoka 812-8582, Japan

福岡市東区馬出３—１—１

Tel: +81-92-642-6030
Fax: +81-92-642-6258

E-mail: 100ko-do@med.kyushu-u.ac.jp
HP: http://med.kyshu-u.ac.jp/100ko-do

Faculty of
MedicalSciences

KyushuUniversity
Hospital
Faculty of
DentalSciences

Entrance

Hall-3

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PROGRAM■ Opening remarks
9:25 – 9:30 Hiroshi Nakanishi, Project leader
9:30 – 9:40 Kazunori Yoshiura, Dean of Kyushu University Faculty of Dental
Science■ Annual Research Reports （9:40 – 11:40）<Chair person: Tamotsu Kiyoshima>(A-1) 9:40 – 9:55
“Angiotensin II mediates the loading signal in human periodontal ligament cells”
S. Monnouchi1, H. Maeda2, S. Fujii2, A. Tomokiyo 2, K. Hori1, and A. Akamine 1,2
1
Division of Oral Rehabilitation, Department of Endodontology and Operative
Dentistry, Faculty of Dental Science, Kyushu University, and 2Department ofEndodontology, Kyushu University Hospital, Fukuoka 812-8582, Japan(A-2) 9:55 – 10:10“Neural stem/progenitor cells in the leptomeningeal niche differentiate into
astrocytes during systemic inflammation”
Zhou Wu, Li Sun, Yoshinori Hayashi, Hiroshi Nakanishi
Department of Aging Science and Pharmacology, Faculty of Dental Science, Kyushu
University, Fukuoka 812-8582, Japan(A-3) 10:10 – 10:25“Fibroblast growth factor 10 and Meckel's cartilage formation in the earlymandibular morphogenesis”
Fumie Terao 1, Ichiro Takahashi1,2, Hidetoshi Mitani1, Naoto Haruyama1,3, Yasuyuki
Sasano1, Osamu Suzuki1, Teruko Takano-Yamamoto 1
1
Division of Orthodontics and Dentofacial Orthopedics, Graduate School of Dentistry,
Tohoku University. 2Department of orthodontics, Faculty of Dental Science, Kyushu
University, Fukuoka 812-8582, 3Tokyo Medical and Dental University, Tokyo113-8549,
Japan(A-4) 10:25-10:40“Interleukin-22-induced STAT3 signal transduction in oral squamous cell
carcinoma”
Lutfun Naher1,2, Tamotsu Kiyoshima1, Seiji Nakamura 2, Hidetaka Sakai1
1

Laboratory of Oral Pathology and Medicine, and 2Department of Oral andMaxillofacial Oncology, Division of Maxillofacial Diagnostic and Surgical Sciences,
Faculty of Dental Science, Kyushu University, Fukuoka 812-8582, Japan
Break 10:40 – 10:55

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<Chair Person: Kanji Tsuru>(A-5) 10:55 – 11:10“Analysis of the molecular mechanisms of periodontal regeneration by EMD”
Takao Fukuda, Terukazu Sanui, Takaharu Taketomi, Takafumi Hamachi,
Katsumasa Maeda
Department of Periodontology, Division of Oral Rehabilitation, Faculty of Dental
Science, Kyushu University, Fukuoka 812-8582, Japan(A-6) 11:10 – 11:25“Studies on characterization of the gene structure and transcriptional regulation of
a novel inositol 1,4,5-trisphosphate binding protein, possibly involved in bonemetabolism”
Ayako Murakami1,2, Miho Matsuda 2, Koushirou Tsutsumi2, Masato Hirata2, andIchiro Takahashi1
1Department of orthodontics, Faculty of Dental Science, Kyushu University
2
Laboratory of Molecular and Cellular Biochemistry, Faculty of Dental Science,
Kyushu University, Fukuoka 812-8582, Japan(A-7) 11:25 – 11:40“Preparation of porous apatite for bone tissue regeneration”
Giichiro Kawachi, Kanji Tsuru and Kunio Ishikawa
Department of Biomaterials, Faculty of dental science, Kyushu University, Fukuoka
812-8582, Japan
Break 11:40 – 12:00■ Nikkiso Co. Inc. presenting Lunchon Seminor<Chair person: Takeshi Yokoyama>
12:00 – 13:00“Eating for better health-Perioperative nutritional management-”
Hideo Iwasaka
Department of Anesthesiology and Intensive Care Medicine, Oita University Faculty of
Medicine, Yufu City, Oita, Japan
Break 13:00 – 13:20■ Special Lecture (SL-1)<Chair persons: Haruyoshi Yamaza / Kazuaki Nonaka>SL-1. 13:20 – 14:20“Stem Cell Property of Post-migratory Cranial Neural Crest Cells and Their
Utility in Tissue Regeneration.”
Yang Chai
Center for Craniofacial Molecular Biology, School of Dentistry, University ofSouthern California, USA
Break 14:20 – 14:30
5■ Special Lectures (SL-2 & SL-3)<Chair person: Toshio Kukita>
SL-2. 14:30 – 15:10“What are the Characteristics of PDL Stem Cells ?”Hidefumi Maeda
Department of Endodontology, Faculty of Dental Science, Kyushu University,
Fukuoka 812-8582, Japan
SL-2. 15:10 – 16:10
Richard O. Oreffo“Skeletal Stem Cells – Bridging the Gap in Bone Regeneration from Cell Biology to
Clinical Translation”
University of Southampton DOHA, Bone & Joint Research Group M887, General
Hospital Hampshire, S016 6YD, UK
Break 16:10 – 16:30■ Special Lectures (SL-4 and SL-5)<Chair person: Ichiro Takahashi>
SL-4. 16:30 – 17:10“Immunomodulatory Properties of Stem Cells from Human Exfoliated Deciduous
Teeth”
Takayoshi Yamaza
Department of Oral Anatomy and Cell Biology, Kyushu University Graduate School of
Dental Science, Fukuoka 812-8285, Japan
SL-5. 17:10 – 18:10“Mesenchymal Stem Cells: From Clinics to Bench-top and Back”
Songtao Shi
Center for Craniofacial Molecular Biology, School of Dentistry,
University of Southern California, USA■ Concluding Remarks
18:10 – 18:30 Drs. Richard Oreffo, Yang Chai, Songtao Shi■ Getting Together Party
19:00 – 20:30

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■ Nikkiso Co. Inc. presenting Lunchon Seminor
Eating for Better Health-Perioperative Nutritional ManagementHideo Iwasaka
Department of Anesthesiology and Intensive Care Medicine, Oita University
Faculty of Medicine, Yufu City, Oita, Japan
食べて健康になるー周術期の栄養管理ー
食べるという文字は人を良くすると書きます。健康であるためには食べることが必須条件になります。近年は過食によるメタボリックシンドロームが注目されていますが、周術期において患者さんは絶食による食事摂取不良が問題となり、周術期の合併症とも関連することが考えられます。低栄養により細胞へのエネルギ
ー供給が低下するとオートファジーと呼ばれる自食作用が発現します。ユビキチ
ンープロテオソーム系が選択的蛋白質分解を行うことに対して、オートファジー
は非選択的に蛋白質や細胞内小器官を分解しエネルギーを得る方法です。基礎的レベルのオートファジーは変性蛋白質の処理機構として必須の機構です。しかし、侵襲とエネルギー供給の低下はオートファジーを過剰に活性化し細胞死をもたらす可能性のあることが指摘されています。周術期の合併症の発現にオートファジーが活性化されることが関連している可能性があります。術前の経口摂取不能に伴うエネルギー不足を補うため私どもの施設では炭水化物含有飲料水を術前２時間まで術前補水食として手術患者さんに提供しています。
手術侵襲はストレス誘導性高血糖をもたらします。高血糖による糖毒性は死亡率の増加と関連することが指摘されています。したがって術中、術後は低栄養をきたさない程度のエネルギー供給と高血糖をもたらさない血糖管理の両面をみた
す管理が大切になってきます。至適なエネルギー投与と高血糖を防止するため人工膵臓による管理が最も安全、効果的管理法と考えられます。私どもの施設での管理方法について説明します。

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■ Special lecture-1
Yang CHAIAFFILIATION

Center for Craniofacial Molecular
Biology, University of Southern California
School of Dentistry

ADDRESSLos Angeles, CA 90033, USA

E-MAILychai@usc.edu

EDUCATION & CAREER
1984

MB, Peking University School of Stomatology, Beijing, China

1984-1986

Teaching Faculty and Surgical Resident, Peking University, Schoolof Stomatology, Department of Oral-Maxilofacial Surgery

1987-1991

Research Associate, School of Dentistry, University of Southern
California

1991-1994

Postdoctoral fellow, Center for Craniofacial Molecular Biology,
University of Southern California

1996-2002

Assistant Professor, School of Dentistry, University ofSouthern California

2002-2006

Associatet Professor, School of Dentistry, University ofSouthern California

2004-2009

Chairman, Division of Craniofacial Sciences and Therapeutics,
School of Dentistry, University of Southern California

2006-Present

Professor, University of Southern California School of
Dentistry

2007-present

Director, Center for Craiofacial Molecular Biology, School of
Dentistry, University of Southern California

2009-present

Associate Dean of Research, School of Dentistry, University ofSouthern California

8Stem Cell Property of Post-Migratory Cranial Neural Crest Cells andTheir Utility in Tissue Regeneration
Yang Chai
Center for Craniofacial Molecular Biology, University of Southern
California School of Dentistry Los Angeles, CA 90033, USA
The vertebrate neural crest is a multipotent cell population that gives rise to a variety ofdifferent cell types. We have discovered that post-migratory cranial neural crest cells(CNCCs) maintain mesenchymal stem cell (MSC) characteristics and show potential
utility for the regeneration of craniofacial structures. We are able to induce the osteogenic
differentiation of post-migratory CNCCs, and this differentiation is regulated by bonemorphogenetic protein (BMP) and transforming growth factor- b (TGF-b) signaling
pathways. After transplantation into a host animal, post-migratory CNCCs form bonematrix. CNCC-formed bones are distinct from bones regenerated by bone marrowmesenchymal stem cells (BMMSCs). In addition, CNCCs support tooth germ survival via
BMP signaling in our CNCC-tooth germ co-transplantation system. Thus, we conclude
that post-migratory CNCCs preserve stem cell features, contribute to craniofacial boneformation, and play a fundamental role in supporting tooth organ development. Thesefindings reveal a novel function for post-migratory CNCCs in organ development, anddemonstrate the utility of these CNCCs in regenerating craniofacial structures.

9■ Special lecture-2
Hidefumi MAEDA
AFFILIATION

Kyushu University Hospital, Department ofEndodontology and Operative Dentistry

ADDRESS

Maidashi 3-1-1, Higashi-ku, Fukuoka,
812-8582, Japan

E-MAILhide@dent.kyushu-u.ac.jp

EDUCATION & CAREER
1984-1990

Kyushu University, School of Dentistry

1990-1994

Kyushu University Graduate School of Dentistry
Ph.D. program in Oral Science

1994-1995

Research Associate, Kyushu University, Faculty of Dentistry,
Department of Pharmacology

1995-1999

Assistant Professor, Kyushu University, Faculty of Dentistry,
Department of Endodontology and Operative Dentistry1999-2001

Research Associate, University of Texas Health Science Center at
San Antonio, Department of Medicine, Division of Hematology,
TX, USA

2001-2003

Assistant Professor, Kyushu University Hospital, Department ofEndodontology and Operative Dentistry2004-present

Lecturer, Kyushu University Hospital, Department ofEndodontology and Operative Dentistry10What are the Characteristics of PDL Stem Cells?
Hidefumi Maeda1, Shinsuke Fujii1, Atsushi Tomokiyo1, Naohisa Wada2,
Satoshi Monnouchi3, Kiyomi Hori3, Katsuaki Koori3, Naohide
Yamamoto3, Akifumi Akamine3
1
Kyushu University Hospital, Department of Endodontology and Operative
Dentistry,
2
University of Adelaide, Colgate Australian Clinical Dental ResearchCentre,
3
Kyushu University, Department of Endodontology and Operative Dentistry,
Fukuoka 812-8582, Japan
Once the periodontal ligament (PDL) tissue is severely damaged by the periodontal
diseases and lost, its reconstruction is well-known to be difficult. Therefore it isrecognized that the regeneration of PDL tissue is the concern for conserving teeth. To
address this issue, researchers have been to try to identify the stem cells, scaffolds, andthe signaling factors responsible for the PDL regeneration. So far we have been focusing
on the stem cells present in PDL tissue. However, as such PDL stem cells are very few toanalyze their characteristics, we developed the immortalized PDL cells derived fromhuman PDL tissue, and cloned these cells. Then we have succeeded in establishing two
clones, 1-11 and 1-17, with multi-differential potentials, capable of characterizing the
PDL stem cells. Line 1-11 contained the potentials to differentiate into osteoblastic cells
and adipocytic cells, while line 1-17 differentiated into osteoblastic cells, adipocytic cells,chondrocytic cells, and neurocytic cells. Furthermore, both cell lines expressed CD13,
CD29, CD44, CD71, CD90, CD105, and CD166 at higher ratios as expressed bymesenchymal stem cells. Therefore using these cell lines we have been evaluating thecharacteristics of PDL stem cells, analyzing the interaction of these cell lines with thescaffold components or the signaling elements.

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■ Special lecture-3
Richard OC Oreffo
AFFILIATION

Bone and Joint Research Group, Center for
Human Development, Stem Cell andRegeneration, Instutute of Developmental
Sciences, University of Southampton

ADDRESS

Southampton, SO16 6YD, UK

E-MAILroco@soton.ac.uk

EDUCATION & CAREER
1980-1983

BSc Hons II I, Biochemistry, University of Liverpool, UK

1983-1986

D.Phil. University of Oxford. (Thesis: “Vitamin A and Bone”)

1987-1989

Research Fellow, Division of Endocrinology, University of Texas,USA

1990-1993

Reswarch Fellow and Principal Research Scientist, Zeneca
Pharmaceuticals, Cheshire, UK

1993-1999

MRC Research Fellow, University of Oxford, Oxford, UK

1999-2002

Non-Clinical Lecturer, University of Southampton, Southampton,UK

2002-2004

Senior lecturer and Reader, University of Southampton,
Southampton, UK

Present

Professor of Musculoskeletal Science Associate Dean Innovationand Enterprise - University of Southampton, UK

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Skeletal Stem Cells – Bridging the Gap in Bone Regeneration from Cell
Biology to Clinical Translation
Richard O. Oreffo
University of Southampton DOHA, Bone & Joint Research Group M887,
General Hospital Hampshire, S016 6YD, United Kingdom
Tissue engineering strategies have sought to repair skeletal defects resulting from trauma
and disease with the application of cells, typically isolated from the patients themselves,in combination with porous biomaterials or scaffolds. Human bone marrow stromal stem
cells or skeletal stem cells are multipotent progenitor cells with the ability to generate
stromal lineages including cartilage, bone, fat and connective tissue. These primitive
progenitors exist postnatally and exhibit stem cell characteristics, namely low incidence
and extensive renewal potential. Advances in our understanding of skeletal stem cells andtheir role in bone development and repair, offer the potential to open new frontiers inbone regeneration.
Our work is centered on isolation, expansion and translational studies of skeletal
populations, including enriched skeletal stem cell populations for skeletal repair using
biomimetic scaffolds together with judicious selection of osteotropic growth factors togenerate appropriate skeletal tissue constructs. A number of areas of work will be
reviewed using data from the group including: i) innovative isolation strategies for of
fetal and adult skeletal populations, ii) derivation of niche environments throughcombination of progenitor cells with tailored biomimetic scaffolds in an attempt tomodulate the osteogenic and angiogenic repair process and iii) translational studies to
examine the efficacy of skeletal populations using impaction bone grafting as an
exemplar. The development of protocols, tools and multidisciplinary approaches that
integrate materials, cell, molecular and clinical techniques for skeletal tissue regeneration
for de novo tissue formation using skeletal progenitor and stem cell populations offers
exciting opportunities to improve the quality of life of many in an increasing ageing
population.

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■ Special lecture-4
Takayoshi YAMAZAAFFILIATION

Department of Oral Anatomy and Cell Biology,
Kyushu University Graduate School of Dental
Science

ADDRESS

Maidashi 3-1-1, Higashi-ku, Fukuoka, 8128582, Japan

E-MAILyamazata@dent.kyushu-u.ac.jp

EDUCATION & CAREER
1988-1994

Kyushu University, School of Dentistry

1994-1998

Kyushu University Graduate School of Dentistry
Ph.D. program in Oral Science

1998- 2001

Research Associate, Department of Endodontlogy and Operative
Dentistry, Kyushu University Graduate School of Dental Science,

2001-2003

Research Associate, Department of Oral Anatomy and Cell
Biology, Kyushu University Graduate School of Dental Science

2005-2006

Special Volunteer, Dental Unit, National Institute of Dental and
Craniofacial Research, National Institutes of Health, Bethesda, MD,
USA, Dr. Songtao Shi's Laboratory

2006-2007

Postdoctoral Research Associate, Center for Craniofacial
Molecular Biology, University of Southern California School of
Dentistry, Los Angels, CA, USA, Dr. Songtao Shi's Laboratory

2008-2009

Research Associate, Center for Craniofacial Molecular Biology,
University of Southern California School of Dentistry, Los Angels,
CA, USA, Dr. Songtao Shi's Laboratory

2003-present

Assistant Professor, Department of Oral Anatomy and Cell
Biology, Kyushu University Graduate School of Dental Science14

Immunomodulatory Properties of Stem Cells from Human ExfoliatedDeciduous Teeth
Takayoshi Yamaza
Department of Oral Anatomy and Cell Biology, Kyushu University Graduate
School of Dental Science, Fukuoka 812-8582, Japan.
Stem cells from human exfoliated deciduous teeth (SHED) have been identified as apopulation of postnatal stem cells capable of differentiating into osteogenic andodontogenic cells, adipogenic cells, and neural cells. Herein we have characterized
mesenchymal stem cell properties of SHED in comparison to human bone marrowmesenchymal stem cells (BMMSCs). Methods: We used in vitro stem cell analysis
approaches, including cytometric flow, inductive differentiation, telomerase activity, andWestern blot analysis to assess multipotent differentiation of SHED and in vivo
implantation to assess tissue regeneration of SHED. In addition, we utilized systemic
SHED transplantation to treat systemic lupus erythematosus (SLE)-like MRL/lpr mice.
We found that SHED are capable of differentiating into osteogenic and adipogenic cells,
expressing mesenchymal surface molecules (STRO-1, CD146, SSEA4, CD73, CD105,
and CD166), and activating multiple signaling pathways, including TGF , ERK, Akt,
Wnt, and PDGF. Recently, BMMSCs were shown to possess an immunomodulatory
function that leads to successful therapies for immune diseases. We examined theimmunomodulatory properties of SHED in comparison to BMMSCs and found that
SHED had significant effects on inhibiting T helper 17 (Th17) cells and promoting
regulatory T cells (Tregs) in vitro. Moreover, we found that SHED transplantation iscapable of effectively reversing SLE-associated disorders in MRL/lpr mice. At thecellular level, SHED transplantation elevated the ratio of Tregs via Th17 cells. These datasuggest that SHED are an accessible and feasible mesenchymal stem cell source fortreating immune disorders like SLE.

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■ Special lecture-5
Songtao SHIAFFILIATION

Center for Craniofacial Molecular
Biology, University of Southern
California School of Dentistry

ADDRESSLos Angeles, CA 90033, USA

E-MAIL

songtaos@usc.edu

EDUCATION & CAREER
1986

MS, Peking University School of Stomatology, Beijing, China

1986-1989

Assistant professor, Peking University, School of Stomatology,
Department of Pediatric Dentistry

1989-1994

Research Associate, University of Southern California, Doheny
Eye Institute Dr. Isaac Bekhor's Laboratory

1994-1997

Postdoctoral fellow, University of California at San Francisco. Dr.
Arnold J. Kahn's Laboratory

1997-1998

Contractor and IRTA fellow, Craniofacial and Skeletal Diseases
Branch, NIDCR/NIH Skeletal Biology Section

1998-1999

Dentist, S&S Best Dental Center, Los Angeles, CA, Clinical
Fellow Craniofacial and Skeletal Diseases Branch, NIDCR/NIH

2002-2003

Clinical Fellow (Supervisor), Craniofacial and Skeletal Diseases
Branch, NIDCR/NIH

2003-2006

Principal Investigator, Craniofacial and Skeletal Diseases Branch,
NIDCR/NIH. Dental Biology Section

2006-2008

Assistant Professor, University of Southern California School of
Dentistry

2008-present

Tenured Associate Professor, University of Southern California
School of Dentistry,16

Mesenchymal Stem Cells: From Clinics to Bench-top and Back
Songtao Shi
Center for Craniofacial Molecular Biology, University of Southern
California School of Dentistry Los Angeles, CA 90033, USA
Mesenchymal stem cells (MSCs) are a population of hierarchical postnatal stem cellswith the potential to differentiate into mesodermal lineage-derived cells including
osteoblasts, chondrocytes, adipocytes, cardiomyocytes, myoblasts and non mesodermal
lineage-derived cells such as neural cells. The orofacial region contains multiple lineage
mesenchymal stem cells, including bone marrow derived MSCs, dental pulp stem cells(DPSCs), periodontal ligament stem cells (PDLSCs), Stem cells from Human ExfoliatedDeciduous teeth (SHED), and Stem Cells from root Apical Papilla (SCAP). First, wefound that MSCs contribute to diseases such as osteoporosis and benign tumors. Next,
we used autologous PDLSCs and periodontal progenitors to cure periodontitis withregeneration of cementum and Sharpey’s fibers in swine and humans. More interestingly,
we used PDLSCs and SCAP to regenerate root/periodontal complexes that are capable ofsupporting artificial porcelain crowns resulting in normal tooth function restoration inswine.

Finally, we generated jaw osteonecrosis model in mouse and employedautologous bone marrow MSC transplantation to cure the necrosis, suggesting a novel
therapeutic approach to recover jaw necrosis by bone marrow MSC-generated new bone
and bone marrow. Our data indicate a great potential of using MSCs to cure a variety ofdisorders and to regenerate functional tissues.

17

■ Annual Research Reports(A-1)

Angiotensin II Mediates the Loading Signal in Human
Periodontal Ligament Cells
S. Monnouchi1, H. Maeda2, S. Fujii2, A. Tomokiyo2, K. Hori1, andA. Akamine1,2
1
Division of Oral Rehabilitation, Department of Endodontology andOperative Dentistry, Faculty of Dental Science, Kyushu University, and2
Department of Endodontology, Kyushu University Hospital, Fukuoka 8128582, JapanThe loading caused by occlusion and mastication plays an important role inmaintaining periodontal ligament (PDL) tissues. We hypothesized that a loading
magnitude would be involved in the production of biological factors that functionin the maintenance of PDL tissues. Here, we identified up-regulated gene
expressions of transforming growth factor- 1 (TGF- 1), alkaline phosphatase(ALP) and angiotensinogen (AGT) in human PDL fibroblastic cells (HPLF) thatwere exposed to 8% stretch loading. Immuno-localization of angiotensin I/II (Ang
I/II), which were converted from AGT, were detected in rat PDL tissues. HPLFs
that were stimulated by Ang II also increased their gene expressions of TGF- 1
and ALP. Furthermore, the antagonist for Ang II type 2 receptor, rather than for
type 1, significantly inhibited gene expressions induced by the stretch loading.
These data suggest that Ang II mediates the loading signal in stretched HPLFs toinduce expressions of TGF- 1 and ALP.

18

(A-2) Neural Stem/Progenitor Cells in the Leptomeningeal Niche
Differentiate into Astrocytes during Systemic Inflammation
Zhou Wu, Li Sun, Yoshinori Hayashi, Hiroshi Nakanishi
Department of Aging Science and Pharmacology, Faculty of Dental Science,
Kyushu University, Fukuoka 812-8581, Japan
Neural stem/progenitor cells (NSCs) are recognized by expression of nestin, anintermediate filament of neuroepithelial derivation. In the adult rat brain, nestin-positive
NSCs are known to exclusively reside in two neurogenic regions, subventricular zone anddentate gyrus of the hippocampus. NSCs can differentiate into both neurons and glial
cells including astrocytes in tight contact with the local environment. Recently, however,
we have found that nestin-positive NSCs also reside in the leptomenigeal niche which
covers the convolusin. By flow cytometrical analyses, nestin-positive NSCs in cultured
leptomenigeal cells from embryonic day 19 rats showed both neuronal and glial spheres.
On the other hand, the nestin-positive astrocytes identified by glial fibrillary acidic
protein (GFAP) were significantly increased after treatment with lipopolysaccharide(LPS). By immunohistochemal analyses, the immunoreactivity for nestin localized in the
leptomeniges of the cerebral cortex and choroid plexus in the young adult rats.
Furthermore, the nestin-positive astrocytes in the proximity of leptomeniges significantly
increased during systemic inflammation caused by adjuvant injection. Moreover, thecultured astrocytes expressed interkeukin (IL)-10 after treatment with LPS. In the
cerebral cortex of the adjuvant injected rats, astrocytes in the proximity of the
leptomeniges also expressed IL-10.
Our observations may aid in understanding the important roles of leptomeninges on
the brain functions during systemic inflammation. We have previously demonstrated thatthe leptomeninges modulate neuronal and glial functions by secretion of diffusible
molecules during systemic inflammation. Now we have provided first evidence thatnestin-positive NSCs in the leptomeningeal niche increasingly differentiate into
astrocytes during systemic inflammation. These differentiated astrocytes from
leptomeningeal NSCs can in turn regulate neuroinflammation by secreting antiinflammatory mediators including IL-10.

19

(A-3) Fibroblast Growth Factor 10 and Meckel's Cartilage Formationin the Early Mandibular Morphogenesis
Fumie Terao1, Ichiro Takahashi1,2, Hidetoshi Mitani1, Naoto Haruyama1,
3, Yasuyuki Sasano1, Osamu Suzuki1, Teruko Takano-Yamamoto1
1
Division of Orthodontics and Dentofacial Orthopedics, Graduate School of
Dentistry, Tohoku University, Sendai, 2Department of orthodontics, Facultyof Dental Science, Kyushu University, Fukuoka 812-8582, 3Tokyo Medical
and Dental University, Tokyo 113-8549, Japan
During organogenesis, fibroblast growth factor (FGF) signaling plays an important role inearly pattern formation and tissue differentiation. In craniofacial development,
differentiation of various tissues and organs involves FGF signaling, such as craniofacialskeletogenesis and mandibular development. However, the role of FGF10 in mandibularmorphogenesis is still unclear. In the present study, we evaluated the role of FGF10 inmandibular chondrogenesis and morphogenesis by using rat mandibular cell micromass
culture and mandibular organ culture in combination with either recombinant FGF10
treatment or electroporation of an FGF10 expression vector, respectively. In mandibularcell micromass culture, FGF10 treatment enhanced chondrogenic differentiation andendogeneous ERK (extracellular signal-regulated kinase) phosphorylation in the cells
derived from lateral area of mandible. In organ culture system, not only the Meckel’s
cartilage was spirally deformed by ectopic expression of FGF10, but also the cartilage
characteristic Col2a1 and Sox9 gene expression were up-regulated. The Meckel’s
cartilage was deformed and the size increased when FGF10 was overexpressed in thelateral area, but not in the medial region of the mandible. In conclusion, FGF10 regulates
the shape and size of Meckel’s cartilage during rat early mandibular morphogenesis
through the differentiation of lateral cells of mandible.

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(A-4) Interleukin-22-induced STAT3 signal transduction in oral
squamous cell carcinoma
Lutfun Naher1, 2, Tamotsu Kiyoshima1, Seiji Nakamura2,
Hidetaka Sakai1
1

Laboratory of Oral Pathology and Medicine, and 2Department of Oral andMaxillofacial Oncology, Division of Maxillofacial Diagnostic and Surgical
Sciences, Faculty of Dental Science, Kyushu University, Fukuoka 812-8582,
Japan
Cytokines or cytokine-related mediators influence the cellular behavior of
malignant tumors. Interleukin (IL)-22 is a member of the IL-10 family. Its main
targets are epithelial cells, but not immune cells. The objectives of this study were
to examine IL-22 signal transduction in oral squamous cell carcinoma (OSCC)
cells. RT-PCR showed that human OSCC cells, MISK81-5 and HSC-3 cells,
expressed IL-22 receptor chains. Immunoblotting showed that IL-22 induced atransient tyrosine phosphorylation of STAT3 (pY705-STAT3) in MISK81-5 cells
after IL-22 stimulation. The change in the serine phosphorylation of STAT3 was
subtle during the examination periods. Meanwhile pY705-STAT3 activation in
HSC-3 cells was undetectable after IL-22 stimulation. The immunocytochemistrydemonstrated that IL-22 induced translocation of phosphorylated STAT3 into the
nucleus of MISK81-5 cells. IL-22 temporarily up-regulated the expression of antiapoptotic and mitogenic genes such as Bcl-x, survivin and c-Myc as well as
SOCS3. IL-22 transiently activated ERK1/2 and induced a delayed
phosphorylation of p38 MAP kinase. IL-22 negligibly involved activation of NFB as detected by using MISK81-5 cells stably transfected with pGL4.26[luc2/minP/Hygro] encompassed four tandem copies of the NF- B consensus
sequence. MISK81-5 cells treated with IL-22 showed mild cellular proliferationand down-regulation of the keratinocyte differentiation-related genes incomparison to unstimulated cells. IL-22 also increased MMP-2 protein in
MISK81-5 cells, but not MMP-9. These results indicated that IL-22 differentially
activated the STAT3 signaling system depending on the type of OSCC. IL-22 may
play roles in the tumor growth, cell differentiation and progression through theactivation of the STAT3 pathway. Modifying the function of IL-22 and/or
blockade of JAK/STAT signals will lead to new perspectives and therapies withrelatively few side effects in order to improve the patient management in OSCC.

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(A-5) Analysis of the Molecular Mechanisms of Periodontal
Regeneration by EMD
Takao Fukuda, Terukazu Sanui, Takaharu Taketomi, Takafumi
Hamachi, Katsumasa Maeda
Department of Periodontology, Division of Oral Rehabilitation, Faculty of
Dental Science, Kyushu University, Fukuoka 812-8581, JapanR (enamel matrix derivative, EMD) is well recognized in
I. Objective: Emdogain ○periodontology, where it is used as a local adjunct to periodontal surgery to stimulate
regeneration of periodontal tissues lost to periodontal disease. However, the exact
molecular mechanisms of periodontal regeneration remain largely unknown. In this study,
we investigated the proteins bound to amelogenin, the major (>95%) component of EMD,
to elucidate the molecules involved in the tissue regeneration during periodontal
treatment.
II. Materials & Methods: We made GST (glutathione S-transferase) fusion amerogeninusing the fetal mouse cDNA, to analyze the proteins bound to amelogenin by GST pull
down assay. Purified GST fusion amelogenin immobilized on glutathione was incubatedwith the lysates of periodontal tissue calls. SaOS-2 and Sa-3, an osteosarcoma cell line
widely used as a model system for osteoblastic cells and human gingival squamous
carcinoma cell line for epithelial cell were mainly used in this study. The cell lysates
were isolated as soluble proteins and membrane proteins respectively. After extensive
washing, bound proteins ware resolved by SDS-PAGE, and analyzed by silver staining orWestern blotting. Thio-His fusion protein was also made to confirm the direct interation
with amelogenin by GST pull down assay.
III. Results: Silver stining analysis showed several proteins bound to amelogenin. Some
proteins bound to amelogenin ware ubiquitous, others ware cell specific. YB-1, human
Y-box protein, was identified as one of the amelogenin binding proteins by the Western
blot analysis.
IV. Conclusion: We identified new amelogenin binding protein, YB-1 (human Y-box
binding protein-1). YB-1 has multiple functions but was initially identified as atranscription factor. YB-1 protein is abundant and expressed ubiquitously in human cells,
functioning in cell proliferation. Our data suggest that YB-1 is involved in regeneration of
periodontal tissues by binding to amelogenin.

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(A-6) Studies on Characterization of the Gene Structure andTranscriptional Regulation of a Novel Inositol 1,4,5-Trisphosphate
Binding Protein, Possibly Involved in Bone Metabolism
Ayako Murakami1,2, Miho Matsuda2, Koushirou Tsutsumi2, Masato
Hirata2, and Ichiro Takahashi1
1Department of orthodontics, Faculty of Dental Science, Kyushu University
2
Laboratory of Molecular and Cellular Biochemistry, Faculty of Dental
Science, Kyushu University, Fukuoka 812-8582, Japan
Phospholipase C-related but catalytically inactive protein (comprising PRIP-1 and -2)
has been isolated as a novel inositol 1,4,5-trisphosphate binding protein with a domainorganization similar to phospholipase C-delta but lacking the emzyme activity. We
have recently reported that PRIP-1 and –2 double knockout (DKO) mice showed
upregulation of gonadotropin secretion, but lower levels of serum sex steroid hormones
were observed. Therefore we analyzed the bone condition of PRIP-DKO mice and theresults indicated the involvement of PRIP in the regulation of bone metabolism. The
PRIP-1 gene is expressed predominantly in the brain, while PRIP-2 exhibits arelatively ubiquitous expression. We speculated that the expression of PRIP-1 in brain
is important for the regulation of hormone balance because central nervous system isinvolved in the regulation of many of hormones secretions.
In this study, we investigated PRIP-1 gene structure and the possible mechanisms
involved in the expression. The tissue distribution pattern of PRIP gene expression inhumans was similar to that in rodents. 5’RACE (rapid amplification of cDNA ends)
analysis using PRIP-1 gene specific primers with human brain mRNA revealed thepresence of three new exons, indicating that the PRIP-1 gene is organized into 8 exonsintervened by 7 introns. Although three transcripts resulting from the alternative
splicing of exon 2 and/or 3 were detected, a transcript lacking exons 2 and 3 waspredominantly expressed in humans, suggesting that the translation start codon ofhuman PRIP-1 exists in exon 1. To characterize the human PRIP-1 promoter, transient
luciferase assay was carried out. The results indicated that the positive regulatory
region is located –237 to –108 bp upstream from the transcription start site. Gel shift
assay revealed the specific binding of some nuclear proteins to this region, suggestingthat the existence of transcription factors contributes to the positive regulation of PRIP1 gene expression. Mutation analyses revealed that the binding of a transcription factor,
MAZ to the regulatory site leads to the promoter activity, indicating that MAZ is
involved in the expression regulation of the human PRIP-1 gene.

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(A-7) Preparation of Porous Apatite for Bone Tissue Regeneration
Giichiro Kawachi, Kanji Tsuru and Kunio Ishikawa
Department of Biomaterials, Faculty of dental science, Kyushu University
Fukuoka 812-8581, Japan
Introduction
For bone tissue regeneration, the materials should have osteoconductivity andbiodegradablity. Autograft is the standard choice for bone tissue regeneration.
Because autograft or bone tissue is absorbed by osteoclasts and then replaced by
new bone. Hydroxyapatite (HAp; Ca 10(PO4)6(OH)2) has been commonly known tohave high biocompatibility and bioactivity [1]. So, conventional bone substitute(HAp) is wildly used in orthopedic and dental fields. HAp shows
osteoconductivity but it never shows biodegradablity. Because HAp withstoichiometric composition (Ca/P atomic ratio = 1.67 can not be absorbed byosteoclasts. So, it could not be replaced with new bone. One of differences
between bone tissue and hydroxyapatite is chemical compositions. Biologicalapatite in bone tissue has a lot of carbonate ion non-stoichiometric composition(calcium deficient composition). We think this is the key for bone substitute. We
already prepared a lot of porous apatite with unique structures by hydrothermal
technique [2, 3, 4, 5].
In this study, we will report the reparation of porous apatite; carbonate apatite
and calcium deficient apatite and discuss its application for bone tissue
regeneration.
Experimental method
As starting material, tricalcium phosphate (TCP; Ca3(PO4)2) and calcium sulfate
were used. These powders were shaped as cylindrical shape of about 8 mmφ ×
2 mmL by molding under 100 MPa. The formed samples were set in a 100 cm3
autoclave with 20 cm3 of NH4Cl / NH3 buffer solution or ammonium carbonate
solution. The pH of solutions was adjusted from 7.3 to 9.5 at room temperature.
Then, samples were treated hydrothermally in liquid phase and vapor phase at 120
or 160 ºC for time up to total 24 h under saturated vapor pressure. Hydrothermalsoaking method means that samples were soaked into liquid and treatedhydrothermally in liquid phase. These buffer solutions were kept same pH volume
after hydrothermal treatment.

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Results
In case of TCP and NH4Cl / NH3 buffer solution, rod-shaped apatite crystals
with about 20 m in length were obtained. Due to crystal growth under thehydrothermal conditions, shape of obtained crystal became rod-shaped.

Fig. 1. XRD pattern and SEM image of apatite crystals prepared hydrotherally in
NH4Cl / NH3 buffer solution (pH7.3) at 160 ℃ for 24 h.
References
1. Hench L. L.: Jouranl of Amerilam Ceramics Society, 81, 1705 (1998).
2. S. Tajima et al.: Dental Materials Journal, 25 [3], 616 (2006)
3. H. Wakae et al.: Journal of Biomedical Material Research A, 87 [4]:957(2008).
4. T. Okuda et al.: Biomaterials, 29, 2719 (2008).
5. G. Kawach et al.: Journal of the Ceramic Society of Japan, 117 [7], 847(2009).

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