The International Joint Symposium on“Dental and Craniofacial Morphogenesis
and Tissue Regeneration”
and“Oral Health Science”
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6 th, February, 2009
Centennial Hall, Kyushu University School of Medicine,
Fukuoka, Japan

Program & Abstracts

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Program                                                          Opening remarks
10:00 - 10:10
Akifumi Akamine (Dean, Faculty of Dental Science, Kyushu Univ.) Special Lectures (Dental and Craniofacial Morphogenesis and Tissue Regeneration)
Chairpersons:SL-1: Kazuaki Nonaka (Section of Pediatric dentistry, Kyushu Univ.)
SL-2: Hiroshi Nakanishi (Laboratory of Oral Aging Science, Kyushu Univ.)

SL-1. 10:10 - 11:10
Non-canonical TGF-b signaling during craniofacial morphogenesis
Yang Chai
Center for Craniofacial Molecular Biology, University of Southern California, Los Angeles, CA, USA
SL-2. 11:10 - 11:50
Glutamate signals in bone
Yukio Yoneda and Takeshi Takarada
Laboratory of Molecular Pharmacology, Kanazawa University Graduate School of Natural Science andTechnology, Kanazawa, Ishikawa, Japan Lunch11:50 - 13:00

 Special Lectures (Oral Health Science)
Chairpersons:SL-3: Yuzo Ninomiya (Section of Oral Neuroscience, Kyushu Univ.)
SL-4: Seiji Nakamura (Oral and Maxillofacial Oncology, Kyushu Univ.)
SL-5: Yoshihisa Yamashita (Department of Preventive Dentistry, Kyushu Univ.)

SL-3. 13:00 - 14:00
Oral sensation and human healthGary K. Beauchamp
Monell Chemical Senses Center, Philadelphia PA, USA
SL-4. 14:00 - 14:40
New Horizon in Oral Science: Driving Force for the Development of Mucosal Immunology
Hiroshi Kiyono
Divison of Mucosal Immunology, Dept. of Microbiology and Immunology,
The Institute of Medical Science, The University of Tokyo, Tokyo, Japan
SL-5. 14:40 - 15:20
Prevention of vascular diseases for aging well
- Periodontal disease is a vascular disease Kenji Matsushita
Department of Oral Disease Research, National Institute for Longevity Sciences,
National Center for Geriatrics and Gerontology, Obu, Aichi, Japan Break
15:20 - 15:30


 Oral Session (Dental and Craniofacial Morphogenesis and Tissue Regeneration)
Chairpersons:OS-1 – 3: Mizuho A. Kido (Oral Anatomy and Cell biology, Kyushu Univ.)
OS-4 – 6: Ieyoshi Kobayashi (Laboratory of Oral Pathology and Medicine, Kyushu Univ.)

OS-1. 15:30 - 15:45
Increased tooth root resorption during orthodontic tooth movement in allergy model rats
Naohisa Murata1,2, Hideki Ioi1, Shunsuke Nakata1 and Mizuho A. Kido2
Departments of 1Orthodontics, 2Oral Anatomy and Cell biology, Graduate School of Dental Science,
Kyushu University, Fukuoka, Japan
OS-2. 15:45 - 16:00
Platelet-derived growth factors is essential for early tooth germ development
Wu Nan1, Tsutomu Iwamoto1, Shinya Yamamoto3, Keigo Yoshizaki1, Akira Sonoda1, Kazuaki Nonaka1 andSatoshi Fukumoto2
1
Section of Pediatric dentistry, Division of Oral Health, Growth & Development Faculty of Dental Science, Kyushu
University, Fukuoka, Japan, 2Division of Pediatric Dentistry, Department of Oral Health and Development Sciences,
Tohoku University Graduate School of Dentistry, Sendai, Japan, 3Department of Pediatric Dentistry Field ofDevelopment, Medicine Course for Health Sciences, Kagoshima University Graduate School of Medical and Dental
Sciences, Kagoshima, Japan
OS-3. 16:00 - 16:15
Functional implication of thymosin beta 4 on the morphogenesis of tooth germ in the mouse lower first molar
Yukiko Ookuma1,2, Ieyoshi Kobayashi1, Tamotsu Kiyoshima1, Kenogo Nagata1, Kazuaki Nonaka2 and Hidetaka Sakai1
1
Laboratory of Oral Pathology and Medicine, Division of Maxillofacial Diagnostic and Surgical Sciences,2
Section of Pediatric Dentistry, Division of Oral Health, Growth and Development, Faculty of Dental Science,
Kyushu University, Fukuoka, Japan
OS-4. 16:15 - 16:30
Wnt signaling and osteoblast differentiation
Etsuko Matsuzaki1, 2, Fumi Takahashi-Yanaga2, Mari Kobayashi1, Shunji Hiratsuka1, Toshiyuki Sasaguri2 andKatsumasa Maeda1
1Periodontology Section, Division of Oral Rehabilitation, Faculty of Dental Sciences, 2Department of ClinicalPharmacology, Faculty of Medical Sciences, Kyushu University, Fukuoka, Japan
OS-5. 16:30 - 16:45
Development of the novel bone augmentation technique-topical application of statinYohei Jinno, Yasunori Ayukawa, Yoichiro Ogino, Ikiru Atsuta and Kiyoshi Koyano
Division of Oral Rehabilitation, Faculty of Dental Science, Kyushu University, Fukuoka, Japan
OS-6. 16:45 - 17:00
Phosphatidylserine liposomes suppress inflammatory bone destruction by phenotypic switching of macrophages
and T cells
Hong Mei Ma1, Zhou Wu1, Toshio Kukita2, Katsumasa Maeda3 and Hiroshi Nakanishi1
1
Laboratory of Oral Aging Science, 2Department of Hard Tissue Regeneration Control & Science,3
Department of Periodontology, Faculty of Dental Sciences, Kyushu University, Fukuoka, Japan Break
17:0017:10


 Oral Session (Oral Health Science)
Chairpersons:OS-7 – 9: Tazuko K. Goto (Department of Oral and Maxillofacial Radiology, Kyushu Univ.)
OS-10 - : Yoshihisa Yamashita (Department of Preventive Dentistry, Kyushu Univ.)

OS-7. 17:10 - 17:25
Detecting the taste cortical region of human using fMRI
Yuko Nakamura1, Tazuko K. Goto1, Kenji Tokumori1, Takashi Yoshiura2, Koji Kobayashi3, Yasuhiko Nakamura3,
Hiroshi Honda2, Yuzo Ninomiya4 and Kazunori Yoshiura1
1
Department of Oral and Maxillofacial Radiology, Faculty of Dental Science, 2Department of Clinical Radiology,
Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan, 3Department of Medical Technology,
Kyushu University Hospital, Fukuoka, Japan, 4Section of Oral Neuroscience, Graduate School of Dental Science,
Kyushu University, Fukuoka, Japan
OS-8. 17:25 - 17:40
Identification of the interaction sites for anti-sweet substances; Gymnemic acid and Grumarin
Keisuke Sanematsu1,2, Noriatsu Shigemura1, Masashi Jotaki1, Seiji Nakamura2, Toshiaki Imoto3 and Yuzo Ninomiya1
1
Section of Oral Neuroscience, and 2Oral and Maxillofacial Oncology, Graduate School of Dental Science,
Kyushu University, Fukuoka, Japan, 3Division of Integrative Physiology, Department of Functional,
Morphological and Regulatory Science, Tottori University, Yonago, Japan
OS-9. 17:40 - 17:55
Globular adiponectin induced secretion of granulocyte colony-stimulating factor in RAW 264 cells
Noriaki Kamio1, Sumio Akifusa1, Noboru Ymaguchi2, Kazuaki Nonaka2 and Yoshihisa Yamashita1
1Department of Preventive Dentistry and 2Department of Pediatric Dentistry,
Faculty of Dental Science, Kyushu University, Fukuoka, Japan
OS-10. 17:55 - 18:10
Relationship between Oral Microbial Flora and Symptoms in Patients with Dry Mouth
Shouichi Shinozaki, Jun-nosuke Hayashida, Masafumi Moriyama, Keiko Hamanaka, Akihiko Tanaka,Haruhisa Yamamoto, Sakae Minami and Seiji Nakamura
Section of Oral and Maxillofacial Oncology, Division of Maxillofacial Diagnostic and Surgical Sciences,
Faculty of Dental Science, Kyushu University, Fukuoka, Japan
 Concluding remarks
18:10 - 18:30
Yang Chai
Gary K. Beauchamp(University of Southern California, CA, USA)
(Monell Chemical Senses Center, Philadelphia PA, USA)

Informations                                                     
Information for Chairpersons
The chairpersons are requested to be the venue by 15 minutes before your session begins.
 Please respect punctuality for the time allocated for presentations and questions/answers in order toensure the smooth progression of the symposium.

Information for Speakers in Special Lectures
Language English
SL-1 and SL-3: 50 minutes are allocated for presentation and 10 minutes for discussion.
 SL-2, SL-4 and SL-5: 35 minutes are allocated for presentation and 5 minutes for discussion.

Information for Speakers in Oral Sessions
Language English
 12 minutes are allocated for presentation and 3 minutes for discussion.

Other details for Presentation
Presentations must be done with your own PC.
The OS should be Windows 2000, or Mac OS 8.5 or upper graded respectively. Your PC must haveexternal monitor output terminal. If the shape of your PC’s external monitor output terminal is special likethe one of SONY VAIO, Mac, etc., please bring your own adaptor for connecting to a D-sub15 pin.
Please prepare your own AC adapter and power cable at the venue for the battery running out.
Please cancel screen-saver and sleep/power-saving settings of your PC monitor beforehand.
Please visit PC Center (near the venue for presentation) during lunch or break times prior to your session tocheck your presentation on the PC, and please hold out your own PC here.
Speakers are requested to operate themselves the PC at the presentation.
If your presentation data contains stills, movies, graphs etc., please be sure to save them and confirm
beforehand that they will play correctly.
Abstracts

SL-1.
Non-canonical TGF-b signaling during craniofacial morphogenesis
Yang Chai
Center for Craniofacial Molecular Biology, University of Southern California,
2250 Alcazar Atreet, CSA 103, Los Angeles, CA 90033, USA
TGF-b/BMP signaling plays a crucial role in an array of biological processes, including embryonicpatterning, organogenesis, wound healing, and carcinogenesis. Previous studies have indicated that
Smad4 serves as the central intracellular effector of TGF-b signaling. In this study, we found that
p38 MAPK is specifically activated by TGF-b and functions as complementary effector inmediating Smad4-independent TGF-b signaling during craniofacial organogenesis. Unlike theBmpr1a mutant, ablation of Smad4 in the dental epithelium does not block early tooth development
and only causes defects in dental cusp patterning and root development. Inactivation of p38 MAPK
signaling in the Smad4 mutant results in the arrest of tooth development at the bud stage, aphenotype identical to that of Bmpr1a mutant mice. Similarly, ablation of Smad4 in the palatalepithelium does not cause cleft palate, but blocking p38 MAPK and Smad4 together does,
reproducing the phenotype of Tgfbr2 mutant mice. Thus, our study demonstrates that p38 MAPK
mediated Smad4-indepenedent TGF-b signaling is a widely used mechanism in regulatingembryonic organogenesis. The ability of epithelium to utilize both Smad4-dependent andindependent pathways to mediate TGF-b signaling illustrates the complicated nature of the TGF-b
signaling mechanism in development and disease.

MemoSL-2.
Glutamate signals in bone
Yukio Yoneda and Takeshi Takarada
Laboratory of Molecular Pharmacology, Kanazawa University Graduate School of Natural Scienceand Technology
The view that L-glutamate (Glu) is an excitatory amino acid neurotransmitter in the mammalian
central nervous system is prevailing on the basis of successful cloning of a number of genes
encoding different signaling molecules, such as Glu receptors for signal input, Glu transporters for
signal termination and vesicular Glu transporters for signal output through exocytotic release.
Although little attention has been paid to an extracellular transmitter role of Glu in peripheral
neuronal and non-neuronal tissues, however, we have for the first time demonstrated the presence ofGlu receptors in peripheral tissues such as adrenal and pituitary glands in 1986. Subsequentmolecular biological and pharmacological analyses including ours give rise to a novel function for
Glu as an autocrine and/or paracrine signal mediator in bone comprised of osteoblasts, osteoclasts
and osteocytes. Functional expression was found with all machineries required for glutamatergic
signaling in osteoblasts, while osteoclasts were devoid of any glutamatergic signaling molecules
except several transporters including the cystine/Glu antiporter. Daily intraperitoneal administrationof Glu significantly prevented the loss of bone mineral density and trabecular bone in both tibia andfemur of ovariectomized mice. Accordingly, Glu could play a dual role in the mechanisms
underlying the maintenance of cellular homeostasis as an excitatory neurotransmitter in the CNS
and as an extracellular signal mediator in peripheral autocrine and/or paracrine tissues. In particular,
we would propose the possible signaling system for Glu to play a role as an extracellular signal
mediator in mechanisms underlying maintenance of the cellular homeostasis in bone. An innovativeand novel interdisciplinary field could be developed on “neuro-osteology” as a scientific bridge
between bone and brain biology in the future.

MemoSL-3.
Oral sensation and human healthGary K. Beauchamp
Monell Chemical Senses Center
Philadelphia PA USA
Oral chemosensory systems (primarily taste and chemesthesis or oral chemical somatosensation)
play a central role in human health. In this presentation I will first briefly describe thesechemosensory systems and then provide a selected overview of a few of their roles. First and mostimportant, these two senses are our nutrient gatekeepers, critical in determining whether a substancewill be taken into the body or rejected. Thus they necessarily are involved in many of the diseasesof excess such as diabetes, hypertension and obesity due to the role of taste in detecting andregulating intake of sugar, salt and perhaps fat. Disturbances of taste, particularly taste loss, whichis relatively uncommon, can be devastating, putting individuals at considerable nutritional risk andmaking them extremely miserable. Much less work has been done on the health-related aspects ofthe chemesthetic system. It is generally assumed that oral irritants that engage this system arousesensations of burn, tingly and pain in order to protect the oral cavity and subsequent stages of thedigestive system from dangerous or toxic compounds. Yet, surprisingly, compounds that stimulate
this system are found in common foods such as hot peppers, menthol products, wasabi, carbonatedbeverages and many others that humans find highly palatable. Recently we have identified an oral
irritant (oleocanthalTM) from extra virgin olive oil that elicits a novel throat-restricted pain that is
considered a very positive sensory attribute. That this compound is also a potent anti-inflammatory
with potential health benefits following long-term consumption may provide a clue as to why it isvalued. Recent research identifying the mechanism by which oleocanthal elicits its burn and studies
on the relation between chemical structure, burn and anti-inflammatory properties also illustrate theintimate connections between oral sensation and human health.

MemoSL-4.
New Horizon in Oral Science:
Driving Force for the Development of Mucosal Immunology
Hiroshi Kiyono
Division of Mucosal Immunology, Department of Mucosal Immunology
The Institute of Medical Science, The University of Tokyo

Oral cavity is the entry of the mucosal immune system (MIS) which is continuously exposed toinfinite antigenic challenges in handling its day-to-day duties. MIS is the first line of immunologicaldefense against invasion of pathogens and hence induces a prompt and robust immune response inorder to avoid invasion of infectious microorganisms. At same time, it also expose to an enormous
number and volume of innocuous and/or instructive antigens which need to be appropriately‘ignored’. Mounting an immunologically harmonized response therefore represents a key
decision-making process of active and/or quiescent responses for the host. The tissues covered bymucosal surfaces including oral and nasal cavities, aero-digestive tract, reproductive tract, middle
year and eye represent a complex immunological network structured to execute theimmunologically harmonized regulation of the two opposite immune responses. Although the areaof mucosal immunology is now highly recognized as one of major streams in the area of medical
and life science, it should be noted that oral science, for example the efforts provided in thedevelopment of caries vaccine in early 1970 triggered the initiation for the concept of the mucosal
immune system. Since oral cavity is an immunologically unique organ consisting with bothmucosal and systemic immune systems, the molecular and cellular understanding of systematic
immune net work in oral cavity will contribute to open up new strategy for the control of infectious
and immunological diseases.

MemoSL-5.
Prevention of vascular diseases for aging well
- Periodontal disease is a vascular disease Kenji Matsushita, D.D.S. Ph.D.
Director, Department of Oral Disease Research, National Institute for Longevity Sciences,
National Center for Geriatrics and Gerontology, Obu, Aichi, Japan
Many changes in the vasculature, hemostasis and endothelium occur during aging. It is
important for aging well to prevent those changes. Nitric oxide (NO) is a second messenger withdiverse roles in the cardiovascular system, such as maintaining continuous vasodilator tone andregulating local perfusion and systemic blood pressure. Impaired endothelial-dependent (NOmediated) vasorelaxation is observed in aging, atherosclerosis, and hypertension. NO is also
inhibiting thrombosis and limiting vascular inflammation. One mechanism by which NO modulates
such disparate physiological processes is by regulating protein trafficking within cells. NO inhibits
exocytosis of endothelial granules which would otherwise trigger inflammation. NO also blocks
platelet secretion of granules that would otherwise activate thrombosis. NO decreases granule
trafficking from the Golgi to the plasma membrane by targeting a key component of the exocyticmachinery, N-ethylmaleimide sensitive factor (NSF). NO regulation of vesicle trafficking is amolecular mechanism that explains some of the cardiovascular effects of NO, and may be of broad
physiological significance. Excessive endothelial exocytosis may play a role in inflammatory andthrombotic disorders such as atherosclerosis, acute coronary syndromes, myocardial infarction, anddeep vein thrombosis. Patients with defective NO synthesis—the hallmark of endothelial
dysfunction—may be at increased risk for acquiring atherosclerosis and coronary events partly
because less NO synthesis enables more exocytosis in response to vascular injury. Drugs directed
at the exocytic machinery of endothelial cells may be useful in decreasing coagulation and vascularinflammation. Peptides that target NSF prolong bleeding time and decrease inflammation in mice.
The exocytic machinery may be a novel drug target for vascular diseases.
Periodontal disease is a disorder that affects about 80% of Japanese adults. Although it isconsidered as an infectious disease, the importance of systemic factors and environmental factors,
such as aging and lifestyle habits, in the etiology of periodontal disease has not been clarified. The
increasing hypercoagulability observed with aging may account for the higher incidence ofthrombotic cardiovascular disorders. Not only impairments of vasculature and gingival tissue by
aging and accumulation of lifestyle habit but also evocation of vascular inflammation and blood
coagulation are also involved in the etiology of periodontitis. In addition, gingipains, which aretrypsin-like proteinases produced by P. gingivalis, activate several blood coagulation factors,
augment exocytosis of endothelial granules, induce expressions of proinflammatory cytokines andmatrix metalloproteases, and then evoke vascular inflammation and blood coagulation. The
cooperative relationship among aging, lifestyle habits, and periodontopathic bacteria is importantfor considering periodontal disease as a vascular disease.

MemoOS-1.
Increased tooth root resorption during orthodontic tooth movement in allergy model rats
1,2

Naohisa Murata, 1Hideki Ioi, 1Shunsuke Nakata, 2Mizuho A. Kido

Departments of 1Orthodontics, 2Oral Anatomy and Cell biology, Graduate School of Dental Science,
Kyushu University.
In the course of orthodontic treatment, tooth root resorption represents an unwanted sequela.
However, the onset and progression of root resorption mechanisms has not been elucidated. In
addition to the factors related to orthodontic treatment (e.g. the duration of treatment or themagnitude of force applied), the patient-related factors (e.g. abnormalities of root morphology orsystemic factors) are also considered to be a risk. Recently we demonstrated using an epidemiologic
approach that orthodontic patients who suffered from an allergic disease such as asthma showed anincreased incident rate of root resorption.
The aim of this study is to determine whether a systemic allergic disease has an adverse effect on
orthodontic tooth resorption. We used Brown-Norway (BN) rats, which are known as high IgEproducers after sensitization that develop late airway response after antigen challenges. Six week
old male BN rats were actively sensitized by a single shot of ovalbumin and aluminium hydroxide
followed by a second shot of ovalbumin after seven days. Seven days after the second sensitization,a stretched closed-coil spring as an orthodontic appliance was inserted between the maxillary left
first molar and the incisors moving the molar mesially with a force of 0.5N. After 7 or 14 days withthe orthodontic appliance, the animals were fixed and were processed for tartrate-resistant acidphospatase (TRAP) histochemistry. The control animal group was also applied with an orthodontic
appliance received no antigen challenge. The pressure zone of the disto-palatal root of the firstmolar was analysed. The area of root resortpion lacuna was wide and deep and the number ofodontoclasts and osteoclasts in the resorption lacuna was significantly elevated in the experimentalgroup compared with that of the control group. Our results suggest that a systemic allergy is one of
the risk factors for root resorption in orthodontic treatment.

OS-2.
Platelet-derived growth factors is essential for early tooth germ development
Wu Nan1, Tsutomu Iwamoto1, Shinya Yamamoto3, Keigo Yoshizaki1, Akira Sonoda1, Kazuaki
Nonaka1, Satoshi Fukumoto2
1. Section of Pediatric dentistry, Division of Oral Health, Growth & Development Faculty of Dental
Science, Kyushu University.
2. Division of Pediatric Dentistry, Department of Oral Health and Development Sciences, Tohoku
University Graduate School of Dentistry
3. Department of Pediatric Dentistry Field of Development, Medicine Course for Health Sciences,
Kagoshima University Graduate School of Medical and Dental Sciences
Tooth development is a complex process that results from a series of sequential and reciprocalinteractions between epithelial and mesenchymal cells. Platelet-derived growth factor (PDGFs) are
known as paracrine growth factors that mediate  epithelial-mesenchymal interactions inorganogenesis. In tooth development, PDGF receptor (PDGFR)- signaling pathway has beenfound to be important for tooth cusp formation. However, the mechanism and function of PDGFs inodontogenesis remain unclear. We found that PDGF-A, PDGF-B, PDGFRand PDGFRmRNA
were expressed from initial stage of tooth development by RT-PCR. Immunofluorescence also
revealed PDGFs expression pattern. PDGF-A and PDGF-B were expressed in both dental
epithelium and mesenchyme. PDGFRwas mainly expressed in dental mesenchyme. Although
PDGFRwas also mainly expressed in dental mesenchyme, its expression was also observed in theenamel knot, which is a signaling center of the tooth that provides positional information for toothmorphogenesis and regulates the formation of tooth cusps. To analyze the function of PDGFs, weestablished tooth organ culture system using the lower first molar tooth germ from E14.5 ICRmouse embryo. Interestingly, in the presence of PDGF-A, 60% tooth germ became bigger than thecontrol ones after 5 days culture. Further, PDGF-A accelerated cusp formation. On the other hand,treatment of AG17, a PDGF receptor tyrosine kinase inhibitor, suppressed the growth and cusp
formation of tooth germ. AG17 also inhibited BrdU positive cells numbers in tooth germ. Thus, ourresults indicate that PDGFs are necessary for initial tooth development, and suggest that PDGF-Aregulate tooth size and cusp formation through controlling cell proliferation.
Key word: PDGF, AG17, proliferation, tooth, mouse.

OS-3.
Functional implication of thymosin beta 4 on the morphogenesis of tooth germ in the mouse
lower first molar
Yukiko Ookuma1,2, Ieyoshi Kobayashi1, Tamotsu Kiyoshima1, Kenogo Nagata1, Kazuaki Nonaka2and Hidetaka Sakai1*
1. Laboratory of Oral Pathology and Medicine, Division of Maxillofacial Diagnostic and Surgical
Sciences, Faculty of Dental Science, Kyushu University, Fukuoka, Japan
2. Section of Pediatric Dentistry, Division of Oral Health, Growth and Development, Faculty of
Dental Science, Kyushu University, Fukuoka, Japan* Research supervisor.
Tooth development is mediated through sequential and reciprocal epithelial-mesenchymal
interactions. There have been many reports regarding the expression of various kinds of genes
which were related in tooth morphogenesis, however the precise signaling pathway has been still
unclear. We previously detected that thymosin beta 4 (Tb4) was a highly expressed gene inembryonic day 12.0 (E12.0) mouse mandible. In this study, we analyzed the functional implicationsof Tb4 in the mouse first molar development, and focused on the effects of Tb4 on the expressionsof matrix metalloproteinase (Mmp), odontogenic genes and nucleolin (Ncl) by using odontogenic
cell lines.
Expressions of the Tb4 mRNA in developing tooth germ were detected by an in situ hybridization.
At E10.5 and E12.0, the expression of the Tb4 mRNA was predominantly observed in thepresumptive dental epithelium. The Tb4 mRNA was expressed in the enamel organ without primary
enamel knot at E15.0. At E18.0, the intense expression was observed in the inner enamel epithelium.
The expression pattern of Tb4 protein was corresponded to that of in situ mRNA expression in thetooth germ. On the basis of theseexpression patterns of Tb4, we performed inhibition assay by using
Tb4 antisense phosphorothioated oligonucleotide (AS S-ODN) in cultured E11.0 mouse mandibles
or E15.0 tooth germs. In addition, we analyzed the mRNA expression of Mmp-2/-9, odontogenic
genes and Ncl by using quantitative real-time PCR in cultured odontogenic cell treated with Tb4 AS
S-ODN. As a result, development of the tooth germ in the organ cultured mandibles at E11.0 was
significantly suppressed by the treatment with the Tb4 AS S-ODN. Enamel and dentin layeringformation was also slightly inhibited in cultured E15.0 tooth germ treated with Tb4 AS S-ODN.
These results indicate that Tb4 plays important roles in the morphogenesis of the developing tooth
germ. The analysis of real-time PCR in the cultured odontogenic cell revealed that the expression of
Mmp-2/-9, odontogenic genes and Ncl mRNAs was markedly suppressed in those cells treated with
Tb4 AS S-ODN. These vitro data may indicate that Tb4 is related to the direct or indirect regulationof transcriptions of Mmp-2/-9, odontogenic genes, Ncl.
The present results demonstrate that Tb4 play important roles in the morphogenesis of the tooth
germ by regulating the expressions of Mmp-2/-9, odontogenic genes and Ncl.

OS-4.
Wnt signaling and osteoblast differentiation
Etsuko Matsuzaki*1, 2, Fumi Takahashi-Yanaga2, Mari Kobayashi1, Shunji Hiratsuka1, Toshiyuki
Sasaguri2, Katsumasa Maeda1
12Periodontology Section, Division of Oral Rehabilitation, Faculty of Dental Sciences,
Department of Clinical Pharmacology, Faculty of Medical Sciences, Kyushu University

Differentiation-inducing factor-1 (DIF-1), a morphogen of Dictyostelium, inhibits cellproliferation and induces cell differentiation in several mammalian cells. We previously reported
that DIF-1 activated glycogen synthase kinase-3b, resulting in the degradation of b-catenin and theinhibition of Wnt/b-catenin signaling pathway. As this pathway has been shown to be involved inbone biology, we investigated the effects of DIF-1 as a negative regulator of this signaling pathway
on SaOS-2 and MC3T3-E1, an osteosarcoma cell line widely used as a model system forosteoblastic cells and murine osteoblast-like cell line, respectively. We found that DIF-1
suppressed the b-catenin protein amount and the activity of the reporter gene containing T-cell
factor/lymphoid enhancer-binding factor (TCF/LEF) consensus binding sites, indicating that DIF-1
inhibited Wnt/b-catenin signaling. DIF-1 also suppressed the expression of osteoblast
differentiation markers, including alkaline phosphatase (ALP), core binding factor a1 andosteocalcin, in a time-dependent manner. Subsequently, we analyzed the effect of DIF-1 on ALP
promoter activity to clarify the mechanism by which DIF-1 suppressed the ALP expression andfound that DIF-1 significantly reduced the promoter activity through the TCF/LEF binding site(-1023/-1017 bp). These results suggest that DIF-1 inhibits ALP promoter activity through the
suppression of Wnt/b-catenin signaling pathway.
Next we examined the effect of anti-Dkk1 antibody and Wnt-3a to investigate the effect of
Wnt/b-catenin signaling activator on osteoblast differentiation. We found that activation of theWnt/b-catenin signaling pathway increased osteoblast mineralization and ALP activity. Therefore,
activator of the Wnt/b-catenin signaling pathway might nave a therapeutic value to induce the boneregeneration.

OS-5.
Development of the novel bone augmentation technique-topical application of statinYohei Jinno, Yasunori Ayukawa, Yoichiro Ogino, Ikiru Atsuta, Kiyoshi Koyano
Division of Oral Rehabilitation, Faculty of Dental Science, Kyushu University
Objectives:
Statins are the drugs for the treatment of hyperlipidemia, and their osteogenic effects have also beenreported. The purpose of this study is to confirm the effectiveness of the locally applied -TCP-collagen gel composite with fluvastatin for the vertical bone augmentation.
Methods:
Fifty-four 10-week-old female rats were used. The composite with (experimental group) or without(control group) 1.0 weight % fluvastatin were percutaneously injected into supra-parietal region.
One, 2, and 4 weeks after the injection, animals were sacrificed and then serum osteocalcin (OC),alkaline phosphatase (ALP), and tartrate-resistant acid phosphatase (TRAP) were measured. Bothhistological and histomorphometrical analyses were also performed.
The test pieces for the analysis of release profile were produced according to the size of injectedcomposite. They were placed into 5ml of saline at 37°C. Once a day, release profiles were analyzed
by UV spectrometry for 4 weeks.
Results:
In the experimental group, bone thickness was significantly thicker than that in control group.
During the period of observation, there were no significant differences in both ALP and TRAP
activities between groups. However, expression level of OC in the experimental group was
significantly higher (ANOVA, P<0.01).
The release amount of fluvastatin from the test piece was maximum at the first day of theexperimental period. From that day, release of fluvastatin continued during the period of
observation.
Discussion:
The release profile of fluvastatin from this composite was similar to that estimated by thehistological data in our previous study. This may result in the increase of both bone thickness andthe serum amount of OC.
Conclusion:
This composite was a potent material for the local delivery of statin to achieve vertical boneaugmentation.

OS-6.
Phosphatidylserine liposomes suppress inflammatory bone destruction by phenotypic
switching of macrophages and T cells
Hong Mei Ma1, Zhou Wu1, Toshio Kukita2, Katsumasa Maeda3,and Hiroshi Nakanishi1
1

Laboratory of Oral Aging Science, 2Department of Hard Tissue Regeneration
Control & Science, 3Department of Periodontology, Faculty of Dental Sciences,
Kyushu University, Fukuoka 812-8582, Japan
We recently found that posphatidylserine-containing liposomes (PS liposomes) inhibit theosteoclast differentiation after phagocytozed by osteoclast precursors. Therefore, PS liposomes canbe implicated as a potential new pharmaceutical intervention against the inflammatory bone
destruction. However, how PS liposomes regulate immune cells infiltration and involve
inflammatory bone destruction remain to be clarified. In the present study, we assess this issue
using inflammatory bone destruction model, adjuvant arthritic (AA) rats. During acute stage of AA
rats, the infiltrated macrophages and helper T cells induced mRNA expression of interleukin(IL)-1b, IL-23, IL-17, transforming growth factor (TGF)-b1 and IL-10 in ankle joints. During
chronic stage of AA rats, mRNA expression of receptor activator of nuclear factor-kB (NF-kB)
ligand (RANKL) and RANK increased significantly in ankle joints, and consistently manifested
bone destruction. Systemic treatment with PS liposomes significantly increased mRNA levels of
IL-10 and TGF-b. In contrast, PS liposomes significantly decreased those of IL-1b, IL-23 andIL-17 without affecting the infiltration of both macrophages and helper T cells during acute stageof AA rats. Furthermore, PS liposomes also decreased mRNA expression significantly in bothRANKL and RANK during chronic stage of AA rats. Moreover, PS liposomes markedly reduced
the bone destruction. Therefore, consisting with the inhibitory effect on osteoclast differentiation,
PS liposomes may also switch macrophages and T cells from pro- to anti-inflammatory phenotype
in turn to inhibit inflammatory bone destruction.

OS-7.
Detecting the taste cortical region of human using fMRI
Yuko Nakamuraa, Tazuko K. Gotoa, Kenji Tokumoria, Takashi Yoshiurab, Koji Kobayashic,
Yasuhiko Nakamurac, Hiroshi Hondab, Yuzo Ninomiyad, Kazunori YoshiuraaaDepartment of Oral and Maxillofacial Radiology, Faculty of Dental Science, Kyushu University,
Fukuoka, Japanb
Department of Clinical Radiology, Graduate School of Medical Sciences, Kyushu University,
Fukuoka, Japanc
Department of Medical Technology, Kyushu University Hospital, Fukuoka, Japand
Section of Oral Neuroscience, Graduate School of Dental Science, Kyushu University, Fukuoka,
Japan
Objectives: fMRI studies of taste tend to decrease specificity of the results by head movements
associated with swallowing. The purpose of this study was to detect the human taste cortical regions
by fMRI using our new system that spreads taste stimuli on the dorsal surface of the tongue as
widely as possible and also prevents swallowing.
Methods: The delivery system of taste solution consisted of an intra-oral and an extra-oral device.
The intra-oral device was assembled with four solution-inlet tubes and an outlet tube that wereattached to an individual mouthpiece. The outlet tube was connected to a continuous suction
apparatus. Healthy volunteers (10 males and 10 females, 19-28 years) participated in this study. The
taste stimuli were sweet (0.5 M sucrose), salty (0.1M sodium chloride), and a tasteless control
solution containing the main ionic components of saliva (25 mM KCl plus 2.5 mM NaHCO3). All
images were acquired with a whole-body 3.0-T MRI scanner. Image processing and data analysis
were performed using the Statistical Parametric Mapping 5 software package.
Results: This device delivered taste solution on most of the dorsum part of tongue and prevented
the swallowing. The head motions of all subjects were less than 2.2 mm in translation and less than
3.0 degrees in rotation in all sessions. As the result of brain mapping, activated area by sweet andsalty taste stimulus located on the putative human primary taste cortices (uncorrected P < 0.001).
These areas were confined and similar to each other.
Conclusion: We detect the human taste cortical regions by fMRI using our new system. This device
could stimulate the subject’s tongue as widely as possible under constant conditions, andsuppressed each subject’s head movements sufficiently.
This system will make it possible to perform precise functional brain mapping with various taste
substances. (300words)

OS-8.
Identification of the interaction sites for anti-sweet substances; Gymnemic acid and Grumarin
Keisuke Sanematsu1,2, Noriatsu Shigemura1, Masashi Jotaki1, Seiji Nakamura2, Toshiaki Imoto3 andYuzo Ninomiya1
1

Section of Oral Neuroscience, and 2Oral and Maxillofacial Oncology, Gracuate School of Dental
Science, Kyushu University, Fukuoka, Japan
3
Division of Integrative Physiology, Department of Functional, Morphological and Regulatory
Science, Tottori University, Yonago, Japan
Gymnemic acid (GA) and Gurmarin (Gur) are a triterpen glycoside and a polypeptide that are
isolated from the plant Gymnema sylvestre, respectively. These chemical compounds are known toselectively suppress taste responses to various sweet substances without affecting responses to salty,sour and bitter substances. Sweet suppressing effect of GA is specific to humans and chimpanzees,
but not to rodents, whereas Gurmarin inhibits the responses to sweet compounds in rodents, but notin humans. It has also been shown that the sweet-suppressing effects of GA and Gur are diminishedby rinsing the tongue with -cyclodextrin (CD) and -CD, respectively.
In order to examine whether GA and Gur directly interact with T1r2+T1r3, we used the sweet
receptor T1r2+T1r3 assay in transiently transfected HEK293 cells. Similar to previous studies inhumans and mice, GA (0.1 mg/ml) inhibited the [Ca2+]i responses of cells heterologously
expressing hT1r2+hT1r3 to SC45647, saccharin and D-tryptophan. The effect of GA rapidly
disappeared after rinsing the cells with 1% -CD. The mouse pair (mT1r2+mT1r3) was notsensitive to GA. Gur (30 g/ml) suppressed the responses of mT1r2+mT1r3-expressing HEK293
cells to various sweetners. The effect of Gur disappeared after application of -CD. The humanpair (hT1r2+hT1r3) was not sensitive to Gur. To identify the interaction sites for GA and Gur, we
examined the responses of the mouse/human chimeras of T1r2 and T1r3. The results suggest thatthe sensitivity to GA depends mainly on the transmembrane region of human T1r3 and Gurinteracts with the extracellular domain of mouse T1r3.

OS-9.
Globular adiponectin induced secretion of granulocyte colony-stimulating factor in RAW 264
cells
Noriaki KAMIO1, Sumio AKIFUSA1, Noboru YAMAGUCHI2, Kazuaki NONAK2, Yoshihisa
YAMASHITA1
1

Department of Preventive Dentistry and 2Department of Pediatric Dentistry, Faculty of Dental
Science, Kyushu University
Adiponectin, an adipocyte-derived cytokine, affects a number of physiological processes, including
immune function and inflammation. Adiponectin may be present as full-length adiponectin (fAd) orconverted to globular domain adiponectin (gAd) by leukocyte elastase, which cleaves the protein’s
N-terminal collagen-like domain. An earlier study demonstrated that the biological activity of gAd
was much stronger than that of fAd. We investigated whether globular adiponectin (gAd) affects theexpression of inflammation-related genes in murine macrophages (RAW 264 cells). Using DNA
microarray analysis, we identified 707 genes with significantly elevated expression levels after 60
min of exposure to 20 g/ml gAd. Of those genes, granulocyte colony-stimulating factor (G-CSF)
showed the most prominent change in expression after 60 min of exposure (104.0 times). The
gAd-induced secretion of G-CSF increased in a time- and dose-dependent manner. U0126 (MEK1/2
inhibitor) and PD98059 (MEK1 inhibitor) reduced gAd-induced G-CSF mRNA expression by 64.5
± 14.0 and 44.2 ± 1.5%, respectively, and they reduced gAd-induced G-CSF protein production by
80.0 ± 2.4 and 46.3 ± 11.9%, respectively. gAd induced the phosphorylation of MEK1/2 and
ERK1/2 in RAW 264 cells. In addition, the gAd-induced phosphorylation of MEK1/2 and ERK1/2
was dramatically reduced by PD98059 and U0126, respectively. Collectively, these results suggestthat MEK1/2-ERK1/2 signaling is involved in the adiponectin-induced secretion of G-CSF.

OS-10.
Relationship between Oral Microbial Flora and Symptoms in Patients with Dry Mouth
Shouichi Shinozaki, Jun-nosuke Hayashida, Masafumi Moriyama, Keiko Hamanaka,
Akihiko Tanaka, Haruhisa Yamamoto, Sakae Minami, and Seiji Nakamura
Section of Oral and Maxillofacial Oncology, Division of Maxillofacial Diagnostic and Surgical
Sciences, Faculty of Dental Science, Kyushu University
ObjectiveDry mouth is characterized by decreased salivary secretion caused by various systemic factors
and side effects of internal drugs. The patients with dry mouth frequently suffer from dental caries,
periodontal diseases and oral mucosal diseases. It has been reported that the oral diseases wereclosely associated with changes in oral microbial flora. In this study, we thus addressed to clarify
the relationship between the oral microbial flora and symptoms in patients with dry mouth.
Materials and Methods
Thirty-nine patients with dry mouth (28 patients with Sjögren's syndrome, 11 patients withdrug-induced or neurotical dry mouth) and 15 healthy controls were studied. Samples wereobtained from water rinsed in their mouths for 30 seconds. We examined clinical findings
including stimulated whole salivary flow rate (SWS), unstimulated whole salivary flow rate (UWS),
number of teeth, and presence of oral mucosal diseases. Oral microbial flora was examined byusing terminal restriction fragment length polymorphism. The number of Candida species wasmeasured after cultivated on CHROMager Candida. Candida species were identified by restriction
fragment length polymorphism after DNA extraction.
Results
In comparison with those in the healthy controls, the patients with dry mouth show thefollowing significant results. The SWS, the UWS, and the number of teeth were decreased. The
presence of oral mucosal diseases in dry mouth was increased. In the microbial analyses,
Actinomyces species and Villonella species in dry mouth were more frequently identified. The
number of Candida species was increased.
Conclusion
There results thus suggest that dry mouth induced by hyposalivation closely associated withdental and oral mucosal diseases, and also with changes in oral microbial flora.