Kyudai Oral Bioscience 2016
-Frontiers in Dental Research and Education in East AsiaFebruary 27th (Sat) 2016
Lecture Theater AB
School of Dentistry Kyushu University

PROGRAM & ABSTRACTS!
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Information for Speakers! Presentation Instruments‘Presentations are restricted to computer presentations using your own personal
computer.
‘Please use a computer running Windows XP, Windows Vista, Windows 7-10,
Machintosh OS X or later and fitted with an external monitor output terminal.
‘All speakers are also requested to bring the data of your presentation on a USB.
Please mark your name and the session number (e.g. S1-1) on the file. Please
make sure that virus check is executed beforehand.

! Making Presentations‘For the speakers in Special Lecture, please bring your own personal computer tothe Computer Operating Desk at 12:40.
‘All speakers are requested to operate the computer by themselves.
‘Your cooperation in finishing your presentation within the allotted time isappreciated.
‘After your presentation, please reclaim your computer from the Computer
Operating Desk.

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PROGRAM3

Kyudai Oral Bioscience 2016 (KOB2016)-Frontiers in Dental Research and Education in East AsiaOpening Remarks
13:00!13:10
Hiroshi Nakanishi (Representative organizer of KOB)
Masato Hirata (Dean, Faculty of Dental Science, Kyushu University )’Session 1“!Dental Research and Education in Vietnam and Japan
Chair person"Kazuaki Nonaka(Section of Pediatric Dentistry)

13:10!13:35 S1-Special Lecture 1
Ngo Thi Quynh Lan
Recent changes in dental education at the Faculty of Odonto-Stomatology, Universityof Medicine and Pharmacy Ho Chi Minh City, Vietnam
Faculty of Odonto-Stomatology, University of Medicine and Pharmacy Ho Chi Minh City, Vietnam

13:35!14:00 S1-Special Lecture 2
Hidefumi Maeda
How can we save a severely-damaged tooth?
Department of Endodontology and Operative Dentistry, Faculty of Dental Science, Kyushu
University, Fukuoka

14:00!14:10# Coffee Break’Session 2“!Dental Research and Education in China and Japan
Chair Person"Zhou Wu(Department of Aging Science and Pharmacology)

14:10!14:35# S2-Special Lecture 3
Wei Xian Yu
The relationship between virulence factors of Porphyromonas gingivalis andperiodontitis
Department of Oral Biomedicine, College and Hospital of Stomatology, Jilin University

14:35!15:00# S2-Special Lecture 4
Xin Wen Zhang
Dental Education and Career Choice between China and Japan
Center of Implant Dentistry, School of Somatology, China Medical University, Shenyang, China

15:00!15:10# Coffee Break4

’Session 3“!Understanding Patients with Mental Disorders
Chair Person: Yoshihiro Tsukiyama (Section of Implant and Rehabilitative Dentistry)
15:10!15:35 S3-Special Lecture 5
Yasunari Sakai
Rare genetic variations and developmental diseases in childhood”
Department of Pediatrics, Kyushu University Hospital

15:35!16:00 S3-Special Lecture 6
Takahiro Kato
Microglia hypothesis of neuropsychiatric disorders from schizophrenia to dementia
translational research
Department of Neuropsychiatry, Graduate School of Medical Sciences, Kyushu University

16:00!16:20# Commemorative Photo & Coffee Break’PhD Student Session (PSS)“
Chair persons"Fumiko Takayama, Yuka Harada and Reiko Yoshimoto
16:20!16:30 PSS-1
Nguyen Thu Thuy
Salivary oxidative stress biomarkers in chronic periodontitis and acutecoronary syndrome
Department of Periodontology, Faculty of Odonto-stomatology, University of Medicine andPharmacy – Ho Chi Minh City, Ho Chi Minh City, Vietnam

16:35!16:45 PSS-2
Shohei Yoshimoto
Hyper-osmotic stress up-regulates proliferation in squamous cell carcinoma cells
Division of General Oral Clinic, Kyushu University Hospital

16:50!17:00 PSS-3
Fan Zeng
Effect of stimulated microgravity on nerve cells
School of Life Science, Beijing Institute of Technology, Beijing, China

17:05!17:15 PSS-4
Xue Li
Brazilian Green Propolis suppress the Porphyromonas gingivalis LPS-induced
inflammatory responses in human fibroblasts by Inhibiting NF-!B activation
Department of Oral Biomedicine, College and Hospital of Somatology, Jilin University, China
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17:20!17:30 PSS-5
Shinichiro Yoshida
Semaphorin 3A induces odontoblastic phenotype in dental pulp stem cells
Department of Endodontology and Operative Dentistry Faculty of Dental Science, Kyushu
University

17:30!17:40# Coffee Break
17:40!17:50 PSS-6
Jie Meng
IL-10 plays an essential role in anti-inflammatory effects of Rheum tanguticum, a
traditional Tibetan medicine, in activated microglia
Department of Aging Science and Pharmacology, Faculty of Dental Science, Kyushu University

17:55!18:05 PSS-7
Masato Dekita
The critical roles of cathepsin S in Porphyromonas gingivalis LPS-induced CD4+ T
cell differentiation
Section of Orthodontics and Dentofacial Orthopedics, Faculty of Dental Science, Kyushu University

18:10!18:20# PSS-8
Chieko Arai
Nephronectin, a basement membrane protein, plays critical roles for differentiationand proliferation of dental epithelial stem cells during tooth development
Section of Orthodontics and Dentofacial Orthopedics, Faculty of Dental Science, Kyushu University

18:25!18:35 PSS-9
Soichiro Sonoda
Patient-derived dental pulp stem cells based regeneration of dentin/pulp-complex
Department of Periodontology, Faculty of Dental Science, Kyushu University

Closing Remarks
18:35!18:40

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ABSTRACTS

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Session 1!
Dental Research and Education in
Vietnam and Japan

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S3-Special Lecture 1
Recent changes in dental education at the Faculty of
Odonto-Stomatology, University of Medicine andPharmacy Ho Chi Minh city, Vietnam
Prof & Dean, Ngo Thi Quynh Lan
1Faculty of Odonto-Stomatology, 2University of Medicine and Pharmacy
Ho Chi Minh city, Vietnam

The Faculty of Odonto-Stomatology, University of Medicine and Pharmacy, HoChiMinh city,
VietNam has established since 1967, is one of the most highly ranked and oldest dental schools in
Vietnam. Its mission is a leading institution in the field of Dental Education and Research through theadvancement of dental knowledge and research, the education and training of dental professionals
towards excellence and the promotion of oral health to the population in Vietnam. The aim of thispresentation to review recent changes in dental education and to orient the revision of current dental
curriculum at the Faculty in future.
The first dental curriculum was established in 1971 with a five-year program including one year for
pre-dental courses, two years for basic Health Sciences and the last two years embracing all aspects ofspecialty in dentistry. In 1975, the curriculum was restructured into a six-year program with additional
subjects (social sciences and maxillo-facial surgery) leading to the Degree of Doctor in
Odonto-Stomatology. In 1979, the first formal educational reform defined the training objectives andthe curriculum of the Faculty of Odonto-Stomatology. in 1989, an important reform was done
according to the Guidelines of the Ministry of Education and Training in order to implement acurriculum which focused on Certificate-based teaching and Preventive and Community Dentistry. It
was called 2.5 + 3.5 program. Then, it was changed 3+3 one in year 2008 and the current program is
2.5 +3.5 now.
In fact, a Mutual Recognition Arrangement (MRA) for Dental Practitioners in the ASEAN

countries which will be implemented in the early of 2016 orients not only the Faculty but also
the dental schools over Vietnam in challenges how to revise the dental curriculum based on theCompetencies of the Dental Professions in country and region. The presentation will also focus
on the issue and raise the open discussion to ask experiences from dental educational leaders in
the regional integration.
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S3-Special Lecture 2
How can we save a severely-damaged tooth?
Hidefumi Maeda
Department of Endodontology & Operative Dentistry, Division of Oral
Rehabilitation,
Faculty of Dental Science, Kyushu University
In Japan facing the super aging society, the government promotes the improvement of healthy life
expectancy. Preservation of tooth and oral health is one of the goals. Along with this, people desire topreserve their teeth as long as possible. Periodontitis and caries had been known to be as major causesof tooth loss. A recent study in Japan, however revealed that the incidence of fracture leading to tooth
extraction was almost the same as that of periodontitis and caries. To save these diseased teeth,
development of novel and unique therapies is required. In this context, we have worked on the issue
how the severely-damaged teeth, in particular, with deep caries and root fracture could be saved. To
solve this problem, we have attempted to approach from the tissue engineering standpoint. In addition
to three classical elements such as stem cells, signal molecules and scaffold, we recognize theinvolvement of neurogenesis and angiogenesis during tissue generation and regeneration. In this
symposium, however I would like to talk about the latest topics associated with above three elements
together with our data, and clinical trials. By integrating these, we believe to develop a novel dental
therapy allowing for longer preservation of such damaged teeth, which will ensure the quality of life inelder people.

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Session 2!
Dental Research and Education in
China and Japan

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S3-Special Lecture 3
The relationship between virulence factors ofPorphyromonas gingivalis and periodontitis
Wei Xian Yu1, Yi Zhang1, Aicaho Gao1, Yanmin Zhou2
1

Department of Oral Biomedicine, 2Department of Dental Implantology,
College and Hospital of Stomatology, Jilin University

Porphyromonas gingivalis (Pg) is known as a keystone pathogen that links to the periodontitis onset
and progression. The bacteria may release a series of virulence factors to evade host defense
mechanisms and impair the host tissue, including lipopolysaccharide (LPS), gingipains, fimbriae,
capsule polysaccharide. We have focused on the pathogenicity of virulence factors of PgLPS on thedestruction of periodontal connective tissue and alveolar bone resorption. On the other hand, the Ephfamily is involved in regulating the process of osteoclast and osteoblast coordination in order tomaintain bone homeostasis. Therefore, we speculate that Eph family can be involved in the restructureof alveolar bone about periodontitis. We have found that PgLPS increased the expression of EphB4
while inhibiting the expression of EphrinB2. Our results demonstrate the EphB4 receptor on osteoblasts
and the EphrinB2 ligand on osteoclasts may generate bidirectional anti-osteoclastogenic andpro-osteoblastogenic signaling into respective cells and potentially facilitate the transition from bone
resorption to bone formation when treated with PgLPS of low concentration (treated with 75 ng/ml for
osteoblast and osteoclast). We have also found that PgLPS increased the expression of EphA2 in bothin osteoblasts and osteoclasts, decreased the expression of osteogenic-related genes and increased theexpression of osteoclast-related genes. In this talk, I will review our researches data to show theimportant roles of Eph family in bone resorption during chronic periodontitis.
These findings may provide information on new targets for prevention and treatment of chronic
periodontitis. I will also introduce the dental education of undergraduate and graduate students in our
College and Hospital of Stomatology, Jilin University, I hope more the activities of education andresearches will be established between our college of Stomatology, Jilin University and the faculty ofdental science, Kyushu University, because one of the partner institutions of our two Universities.

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S3-Special Lecture 4
Dental Education and Career Choice between China
and Japan
Xin Wen Zhang1, Xu Yan2
1

Center of Implant Dentistry, School of Stomatology, China Medical

University, Shenyang, China; 2 The VIP department, School of Stomatology,
China Medical University, Shenyang, China.
Dependent on our dental education background in Japan as the foreign students and our dental
education career in China as the teachers, we have noticed that there are large differences in dental
education and career choice of dental students between China and Japan. Because of the interest, we
initiated design self-answered questionnaires and investigate differences and similarities among thedental students in the two nations in 2013. We have found that from the very beginning, the motivations
to choose dentistry were highly distinct. Japanese dental students mainly based on the reasons ofhelping others, achieving self- worth and health-care related interests. On the other hand, most of
Chinese students pointed out that their choices were for the financial and prestige. Even approximate
one-third of them signified that dentistry were not their original aspiration just regulated from othersubjects. During the undergraduate phase, the rate of the satisfaction of teaching faculties in Japan was
3 times compared to China. More than two-thirds Chinese students wondered they wasted too muchtime in studying foreign language, while there is no English necessary condition for graduation in
Japan Higher Education. After graduated, most of Chinese students chosen to be further educatedbecause of the great employment pressure, while the rate of Japanese students was less. Furthermore,
more than a half of the graduates prefer work in university hospitals in China, while the majority ofJapanese graduates chose to be general dentists and work in dental offices.
In this talk, we will review our dental education and researches in Japan and introduce the dental
education in China. I hope the information will help to realize the students’ requirement of distinct
cultural and social background. As foreign students, and as graduates of dental school, Kyushu
University, we will do our best to work as a bridge of dental education between China and Japan.

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References:
1. Xu Yan, Xinwen Zhang, Yong Shen, Zhe Yi, Lin Wu, Hongjun Ai. Career choice and future plan ofChinese 8-year stomatology medical doctor program students Journal of the Chinese Medical
Association. (2015) 78: 555-561.
2. Xu Yan, Xinwen Zhang, Yohei Jinno, Keishu Tachibana, Jie Gao, Kiyoshi Koyano, Hongjun Ai,
Yong Shen. Career choice and future design of dental students in China and Japan. International
dental journal (2014) 64: 68-75.
3. Xu Yan, Xinwen Zhang, Yohei Jinno, Keishu Tachibana, Jie Gao, Kiyoshi Koyano, Yong Shen,
Hongjun Ai. Comparison of attitudes towards dental education among dental students in Japan and
China. International dental journal. (2014) 64:76-82.
Acknowledgment
The studies are supported by National Natural Science Foundation of China to Xinwen Zhang(NO.81500858).

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Session 3!
Understanding Patients with
Mental Disorders

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S3-Special Lecture 5
Rare genetic variations and developmental diseases inchildhood
Yasunari Sakai, M.D., Ph.D.
Department of Pediatrics, Kyushu University

In the last decade, physicians have intimately collaborated with geneticists to investigate the genetic
backgrounds underlying developmental disorders in childhood. In particular, whole-exome sequencing(WES) technologies enable us not only to understand the evolutional process of human genome, but it
also provides the chance for identifying rare pathogenic mutations in developmentally challenged
children. The latter include copy number alterations and single-nucleotide variations in individuals with
autism and other forms of neurodevelopmental disorders. Notably, these genetic variations are known
to occur de novo in probands at significantly higher rates than healthy individuals. Massive data with
the WES strategies for thousands of families in the United States and Europe also revealed that
hundreds of genes were likely involved in the pathogenic processes of autism and infantile-onset
epilepsies. On the other hand, it remains to be discussed whether the second and third variations inaffected individuals may be associated with their atypical or severe phenotypes. I herein present several
cases carrying rare mutations and multiple genetic burdens that were considered to cause unusual
phenotypes. Experiences from these cases support the concept that genome-wide scanning of rarevariations will help identifying common therapeutic targets in future translational medicine.

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S3-Special Lecture 6
Microglia hypothesis of neuropsychiatric disorders fromschizophrenia to dementia - translational research
Takahiro A. Kato, MD, PhD1, 2 , Masahiro Ohgidani, PhD1,
Motoki Watabe, PhD3, Shigenobu Kanba, MD, PhD1
1

Department of Neuropsychiatry, Graduate School of Medical Sciences,

Kyushu University, Fukuoka, Japan ,2Innovation Center for Medical Redox Navigation, Kyushu
University, Fukuoka, Japan, 3Monash University, School of Business, Sunway, Malaysia
Microglia have been implicated in various neurological and psychiatric disorders in rodent andhuman PET and postmortem studies. However, the dynamic actions of microglia in the living humanbrain have not been clarified due to a lack of studies dealing with in situ microglia. We introduce anovel technique for developing induced microglia-like (iMG) cells from human peripheral blood cells.
An optimized cocktail of cytokines, GM-CSF and IL-34, converted human monocytes into iMG cells
within 14 days. The iMG cells have microglial characterizations; expressing markers, forming ramified
morphology, and phagocytic activity with various cytokine releases. Clinical utilities were confirmed
by iMG cells from a patient of Nasu-Hakola disease (Ohgidani et al. 2014). On the other hand, how
microglial activities contribute to human psychosocial aspects has not been well clarified. We haverecently conducted an economic game – trust game – to healthy young males with or without
4-day-treatment of minocycline, an antibiotic with microglial inhibitory effects. Interestingly,
minocycline suppressed personality-oriented and/or drive-oriented social behaviors (Watabe et al.
2013). These results have proposed novel psychosocial roles of human microglia.
We believe that the iMG-technique and the pharmacological trials with minocycline promise to
elucidate unresolved aspects of human microglia in various neurological and psychiatric disorders.
References
Watabe M*, Kato TA*, Tsuboi S, et al.: Minocycline, a microglial inhibitor, reduces ‘honey trap’ risk inhuman economic exchange. Scientific Reports, 3, 1685, 2013
Ohgidani M, Kato TA*, Setoyama D, et al.: Direct induction of ramified microglia-like cells fromhuman monocytes: Dynamic microglial dysfunction in Nasu-Hakola disease. Scientific Reports, 4,4957, 201417

Ohgidani M, Kato TA*, Kanba S: Introducing directly induced microglia-like (iMG) cells from fresh
human monocytes: A novel translational research tool for psychiatric disorders. Frontiers in
Cellular Neuroscience, 9, 184, 201518

PhD Student Session

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PSS-1
Salivary oxidative stress biomarkers in chronic periodontitis and acutecoronary syndrome
Thuy T. Nguyen1,2, Lan Q. Ngo3, Ananya Promsudthi4, Rudee Surarit2
1

Department of Periodontology, Faculty of Odonto-stomatology, University of Medicine andPharmacy – Ho Chi Minh City, Ho Chi Minh City, Vietnam, 2Department of Oral Biology, Facultyof Dentistry, Mahidol University, Bangkok, Thailand, 3Department of Dental Basic Sciences,
Faculty of Odonto-stomatology, University of Medicine and Pharmacy – Ho Chi Minh City, Ho Chi
Minh City, Vietnam, 4Department of Oral Medicine and Periodontology, Faculty of Dentistry,
Mahidol University, Bangkok, Thailand
ABSTRACT
Objectives: The study aimed at evaluating oxidative stress (OS) biomarkers in the saliva of patientswith chronic periodontitis (CP) and acute coronary syndrome (ACS) and establishing their correlation
to periodontal index and cardiovascular markers.
Methods: The present study enrolled 96 patients divided into four sex and age-matched groups asfollows: ACSCP group (with both CP and ACS), ACS group (with ACS only), CP group (with CP only),and group of healthy controls. Plaque index (PI), gingival index, bleeding on probing, probing pocket
depth (PPD), and clinical attachment loss were recorded. Cardiovascular markers included serum highsensitivity C-reactive protein and plasma fibrinogen. The levels of OS biomarkers related to lipid,
DNA/RNA and protein oxidation were measured in saliva samples using ELISA andspectrophotometric assays.
Results: Salivary OS biomarkers in diseased subjects expressed statistically significant differences inlevels compared to healthy controls (p<0.05). There were significant correlations between some OS
biomarkers and periodontal parameters as well as biomarkers for cardiovascular events (p<0.05).
Conclusion: Salivary OS biomarkers could potentially serve as diagnostic tools for cardiovascular
and/or periodontal diseases.

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PSS-2Hyper-osmotic stress up-regulates proliferation in squamous cell
carcinoma cells
Shohei Yoshimoto1, Hiromitsu Morita2, Yoshinori Katakura3,
Miho Matsuda1, Seiji Nakamura4, Masato Hirata1
1

Lab. of Mol. & Biochem., Fac. of Dent. Sci., Kyushu Univ., Fukuoka, Japan, 2Sect. of General
Dentistry, Dept. of General Dentistry, Fukuoka Dental College, Fukuoka, Japan, 3Fac. ofAgriculture, KyushuUniv., Fukuoka, Japan, 4Sect. of Oral and Maxillofac. Oncol., Div. of
Maxillofac. Diag. Surg. Sci., Fac. of Dent. Sci., Kyushu Univ., Fukuoka, Japan

Epidermal growth factor receptor (EGFR) is highly expressed and plays an essential role for cancer
progression in oral squamous cell carcinoma. Therefore, EGFR is recognized as a molecular target inoral cancer therapy. On the other hand, tumor microenvironment is constantly inflamed, and it could bepossible that the hyper-osmotic stress, given by inflammation, is involved in cancer proliferation. Here
we report that hyper-osmotic stimulation to human tongue squamous cell carcinoma cells (HSC-3)
triggered activation of nuclear factor of activated T-cells 5 (NFAT5), followed by subsequent activationof a regulator protein of N-glycosylation, dolichol phosphate-dependent N-acetylglucisamine
1-phospho-transferase (DPAGT1), thus resulting in translocation of EGFR by glycosylation from thecytosol to the plasma membrane.
EGFR were translocated to the plasma membrane by changing medium from normal (D-MEM; 300mOsM) to hyper-osmotic condition (D-MEM plus 50 mM mannitol; 350 mOsM), as assessed byfluorescent immunocytochemistry and Western blotting analysis. Cell proliferation assays (WST-8 and
BrdU uptake) revealed that the growth of HSC-3 cells was accelerated by hyper-osmotic stimulation,
but suppressed by CP380736 (10 µM), an EGFR selective inhibitor or genistein (10 µM), a potent
tyrosine kinase inhibitor. In addition, EGFR was accumulated in the endoplasmic reticulum assessedby immunofluorescent observation, when cells were cultured in normal osmotic condition, but withlow glucose concentration. The accumulation was diminished in hyper-osmotic condition even withlow glucose concentration. Furthermore, HSC-3 cells whose NFAT5 was knocked down by shRNA,
showed that the events triggered by hyper-osmotic stimulation were significantly suppressed.
Interestingly, expression of DPAGT1 was also decreased. Chromatin immunoprecipitation assay (ChIP
assay) revealed the binding of NFAT5 to the promoter region of DPAGT1 in hyper-osmotic condition.
These results indicate that NFAT5 participates in HSC-3 cell proliferation by promoting thetranslocation of EGFR to the plasma membrane, triggered by glycosylation by DPAGT1 in
hyper-osmotic condition.
Keywords : EGFR, NFAT5, Hyper-osmotic
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PSS-3
Effects of simulated microgravity on neuronal cells
Zeng Fan, Wang Xianghan, Wang Demei, Qing Hong*
School of Life Science, Beijing Institute of Technology, Beijing 100081, China
Microgravity is a crucial factor during space flight that affects the healthy and work efficiency ofastronauts. In the present study, we investigated whether simulated microgravity could influencemorphology and apoptosis of different regions of brain. Explanted tissues and primary cells wereobtained from three regions of neonatal SD rats, namely hippocampus, cerebellum and cortex. After
cultured for 7 days at 37 $, 95 % of humidity and 5 % CO2, cultures were randomly divided into
control group (Ctr) and simulated microgravity group (sMG). The sMG group was placed on clinostat
for rotating and the Ctr group was cultured on the same horizontal line of the clinostat. After treated for
1 day, 7 days and 14 days for both two groups, changes of morphology and apoptosis of the three
regions were detected by microscope and flow cytometry. Results showed that after treated with seriesof time of simulated microgravity, the surface of explanted tissues changed a lot compared to Ctr,
especially the edge of tissue blocks became brighter. And the cells within explanted tissues became less.
But apoptosis of primary cells of the three regions showed no difference after 1-day and 7-day rotating,while after 14 days of stimulating by simulated microgravity the apoptosis rate decreased dramatically.
Also, human glioma tissues which were offered by Xuan Wu Hospital were cut into explanted tissue
blocks. The procedure was totally the same with that of SD rats and results indicated that cytoskeleton
was influenced by simulated microgravity.
In conclusion, simulated microgravity had the same impact on morphology and apoptosis of the three
regions of brain, which were hippocampus, cerebellum and cortex. And the cytoskeleton of glioma cells
could be disorganized in the simulated microgravity environment.
*Correspondence: Qing Hong ”QH) hqing@bit.edu.cn

Acknowledgment!!
This study was supported by the National Key Foundation for Exploring Scientific Instrument of China(2012YQ04014008, 2013YQ03059514).

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PSS-4
Brazilian Green Propolis suppress the Porphyromonas gingivalis
LPS-induced inflammatory responses in human fibroblasts by Inhibiting
NF-!B activation
Xue Li1, Zhou Wu2, Junjun Ni2, Hiroshi Nakanishi2, Yanmin Zhou1
1

Department of Implantology, Jilin University, China, 2 Department of Aging Science andPharmacology, Kyushu University, Japan

Periodontitis, as a chronic systemic inflammation, has been recognized positive link to cognitive
impairment in Alzheimer’s disease. Therefore, the management of periodontitis might be one of theapproaches for preventing cognitive impairment. Fibroblasts are the resident cells in the connective
tissues including gingival, which is the principal source of the extensive extracellular matrix (ECM)
throughout the body. In dental area, gingival fibroblasts are consisted of oral epithelium that!forming
epithelium attachment for teeth as well as the dental implants during the teeth lost. Regulating of
gingival fibroblasts are important for the managements of periodontitis and Peri-implantitis, because
the gingival fibroblasts are involved in the procedure of inflammatory responses by the mediator
production. Porphyromonas gingivalis (Pg) the keystone oral bacteria, because Pg andlipopolysaccharides from Pg (PgLPS) are involved in the process of periodontitis as well as
Peri-implantitis. Considering the fact that propolis, the nature production from bees, have anti-oxidative
and anti-inflammatory effects, we hypothesized that propolis may have protective effects against the
PgLPS-induced inflammatory responses in human fibroblasts. In the present study, we firstly
demonstrated the expression of Toll-like receptor 2 (TLR2), TNF-", IL-1# and IL-6 in the gingival
fibroblasts of chronic periodontitis patients. In in vitro studies, PgLPS (1 µg/ml) increased the meanmRNA levels of TLR2, TNF-", IL-1# and IL-6 in BJ human fibroblast cell line. The PgLPS-increased
the mRNA levels of TLR2 as well as TNF-", IL-1# and IL-6 in human fibroblasts significantly
decreased by pre-treatment with Bay 11-7082 (20 µM), a specific inhibitor of NF-!B. Moreover,propolis significantly decreased the PgLPS-increased NF-!B activation as well as the expression of
pro-inflammatory cytokines in human fibroblasts.
These observations provide the first evidence that propolis inhibit the PgLPS-induced inflammatory
responses in fibroblasts, thus suggest that propolis may be benefit for management of oral health anddental implants.
Acknowledgment
This study is partly supported by Yamada Research Grant to Zhou Wu (N0.0183), Xue Li is supported
by Kyushu University Friendship Scholarships (2015-2016).

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PSS-5
Semaphorin 3A induces odontoblastic phenotype in dental pulp
stem cells
Shinichiro Yoshida1, Naohisa Wada2, Daigaku Hasegawa3, Atsushi Tomokiyo3,
Sayuri Hamano3, Hiromi Mitarai1, Suguru Serita1, Hiroyuki Mizumachi1,
Hidefumi Maeda1, 3.
1

Department of Endodontology and Operative Dentistry, Faculty of Dental Science, Kyushu
University, 2Division of General Dentistry, Kyushu University Hospital, and 3Department ofEndodontology, Kyushu University Hospital

Exisiting pulp capping materials possess low potentials to accelerate the biological activities ofdental pulp cells, and have a limited capacity to reconstruct dentin/pulp complexes. Therefore, thedevelopment of more effective therapeutic agents for direct pulp capping has been anticipated. Here,
we investigated the effects of Semaphorin 3A (Sema3A) on various functions of human dental pulp
stem cells (DPSCs) in vitro and reparative dentin formation in a rat dental pulp exposure model in vivo.
The expression of Sema3A and its receptors in human DPSCs was examined by semi-quantitative
RT-PCR, immunofluorescence staining and western blotting analysis. The effects of Sema3A on cell
migration, chemotaxis, proliferation and odontoblastic differentiation of DPSCs were examined byscratch wound healing assay, transwell assay, WST-1 proliferation assay, quantitative RT-PCR andAlizarin Red S staining, respectively. The effect of Sema3A application by direct pulp capping on
reparative dentin formation was assessed using a rat pulp exposure model. Nuclear accumulation of#-catenin, the expression of FARP2 gene and activated Rac1 in Sema3A-treated DPSCs were examinedby immunofluorescence staining, western blotting analysis, quantitative RT-PCR and Rac1 pull-down
assay, respectively.
Sema3A and its receptors were expressed in rat dental pulp tissues and human DPSCs. Sema3A
promoted cell migration, chemotaxis, proliferation and odontoblastic differentiation of human DPSCs.
In a rat pulp exposure model, Sema3A facilitates reparative dentin formation with dentin tubules andwell-aligned odontoblast-like cell layer after direct pulp capping treatment. Exogenous application of
Sema3A to human DPSCs increased nuclear accumulation of #-catenin, and also upregulated theexpression levels of FARP2 gene and activated Rac1.
Sema3A plays crucial roles in various events associated with dentin regeneration, probably via
canonical Wnt/#-catenin signaling. Although further study is needed, Sema3A might be an alternative
agent for direct pulp capping.
24

PSS-6
IL-10 plays an essential role in anti-inflammatory effects ofRheum tanguticum, a traditional Tibetan medicine, in activated microglia
Jie Meng1, Aiqin Zhu2, Hiroshi Nakanishi1, Zhou Wu1
1

Department of Aging Science and Pharmacology, Kyushu University, Fukuoka 812-8582, Japan,
2

Institute of Geriatric Qinghai Provincial Hospital, Xining 810007, China

Over the past decade, numerous lines of evidence have strongly suggested that chronic
neuron-inflammation characterized by the release of pro-inflammatory mediators from hyper-activatedmicroglia plays a critical role in the pathogenesis of cognitive dysfunction associated with Alzheimer’s
disease (AD). Basic researches provide substantial evidence that the development of agents that canregulate microglial activation has been suggested as one potential strategy for the treatment or
prevention of cognitive dysfunction associated with AD. Rheum tanguticum Maxim. ex Balf., a
traditional Tibetan medicine, is known to exhibit various bioactivities, including anti-bacteria,
anti-inflammatory and anti-oxidant activities. Therefore, Rheum tanguticum may have the potential to
be used as pharmacological therapeutic agent in treatment with AD. In this study, we have thus
attempted to elucidate effects of Rheum tanguticum on pro-inflammatory effects in activated microglia.
Rheum tanguticum significantly inhibited the mRNA expression of pro-inflammatory molecules,
including interleukin-1!, tumor necrosis factors-" and inducible nitric oxide synthase in microglia after
treatment with chromogranin A, an endogenous microglial activator. This anti-inflammatory effect ofRheum tanguticum was suppressed by the inhibition of the NF-#B and STAT-1 activation pathways.
Interestingly, Rheum tanguticum significantly increased the mRNA expression of interleukin-10 (IL-10),
an anti-inflammatory cytokine with important roles in preventing inflammation, even in non-stimulated
microglia. Furthermore, Rheum tanguticum significantly increased the mRNA expression of myocyte
enhancer factor 2D (MEF2D), a potential transcription factor of IL-10 gene, in microglia.
These results suggest that Rheum tanguticum has anti-inflammatory effects in activated microglia
through increased expression of IL-10.
Key words: microglia, Rheum tanguticum, anti-inflammatory effects, IL-10, MEF2D

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PSS-7The critical roles of cathepsin S in Porphyromonas gingivalis
LPS-induced CD4+ T cell differentiation
Makoto Dekita1, Zhou Wu2, Junjun Ni2, Hiroshi Nakanishi2, Ichiro Takahashi1
1

Section of Orthodontics and Dentofacial Orthopedics and 2Department of Aging Science andPharmacology, Kyushu University, Fukuoka 812-8582, Japan’Background and Objective“There is accumulating evidence that periodontitis is exacerbating risk
factor for numerous of diseases, including Alzheimer’s disease. Low-grade chronic systemic
inflammatory responses associated with periodontitis may activate primed or senescent microglia,
which subsequently provoke an exaggerated neuroinflammation, which may lead to the cognitive
impairment. It is considered that oral bacteria inducing periodontal disease and their virulence factors
can amplify the host immune responses after entering systemic circulation. Porphyromonas gingivalis(Pg) is well known as a keystone bacterium in periodontitis because it mainly responsible for the onset
and progression of periodontitis. Therefore, we hypothesize that lipopolysaccharide from Pg (PgLPS)
may amplify the host immune responses in the spleen, the largest lymphatic organ, because we haverecently reported that the spleen is involved in the amplification of abnormal chronic pain throughcathepsin S (CatS)-dependent differentiation of CD4+ helper T cells (Zhang et al., J. Neurosci., 2014).
In the present study, we have thus attempted to clarify possible roles of CatS in PgLPS-induced
immune responses in the spleen.
’Methods and Results “In the in vivo experiments, a significant hypertrophy of the spleen was
found in the wild-type (female DBA mice with12-week old), but not in CatS-/- mice, after systemictreatment with PgLPS (100 µg/mg/day, i.p.) for 7 consecutive days. Rather surprisingly, CD11c+
dendritic cells (CDs), which expressed IL-6, were significantly increased in the spleen of wild-type, but
not CatS-/-, mice. In the in vitro experiments, PgLPS (1 µg/ml, 24 hs) induced the IL-6 production in
DCs acutely isolated from the spleen of wild-type, but not CatS-/-, mice. The IL-6 production after
treatment with PgLPS was completely suppressed by a pretreatment with Z-Phe-Leu-COCHO (1µM), a
selective inhibitor of CatS.
’Conclusion“These observations indicate that CatS plays a critical role in the differentiation of
Th17 cell in the spleen following chronic systemic treatment with PgLPS. Further studies are needed toelucidate the precise mechanism underlying CatS-mediated IL-6 production in DCs, because IL-6 is thenecessary factor for the differentiation of Th17 cells.
Key words: Porphyromonas gingivalis, LPS, cathepsin S, dendritic cells, IL-6, Th17 cells
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PSS-8Nephronectin, a basement membrane protein, plays critical roles fordifferentiation and proliferation of dental epithelial stem cells during
tooth development.
Chieko Arai , Keigo Yoshizaki , Kanako Miyazaki , Xue Han , Keita Funada ,Ichiro Takahashi
Section of Orthodontics and Dentofacial Orthopedics, Kyushu University, Fukuoka, Japan
Tooth morphogenesis is initiated by reciprocal interactions between epithelial and mesenchymal cells.
Basement membrane (BM) lying between epithelia and mesenchyme plays important roles for thosesignaling interactions. To identify the functions of BM in tooth development, we focused on an
extracellular matrix protein, Nephronectin (Npnt), which is highly expressed in organs of whichdevelopment is largely dependent upon epithelial-mesenchymal interactions.
In RT-qPCR assay, Npnt is highly expressed in tooth, lung and kidney during their embryonicmorphogenesis, especially, strongly during the early stages of tooth morphogenesis. It is reported that
Npnt deficient mice showed kidney agenesis, suggesting that Npnt is also important for toothdevelopment. As we expected, the size of tooth germ was significantly smaller than that of the controls
in presence of Npnt siRNA in organ culture system. Immunohistochemistry showed that Npnt localized
in BM in E11, E13 and E14 molar tooth germ. Near to the cervical loop of P1 incisor, immunostainingof Npnt showed inversed expression pattern of Sox2, which is dental epithelial stem cell marker.
Consistently, number of Sox2 positive (Sox2+) cells was significantly reduced by overexpression of
Npnt, while the proliferation increased in vitro.
These results indicated that Npnt localizes in BM, and regulates the differentiation and proliferationof SOX2+ dental epithelial cell during tooth development.

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PSS-9
Patient-derived dental pulp stem cells based regeneration ofdentin/pulp-complex
Soichiro Sonoda1, 2, Takayoshi Yamaza2, Fusanori Nishimura1
Departments of 1Periodontology and 2Molecular Cell Biology and Oral Anatomy, Kyushu University
Graduate School of Dental Science, Fukuoka, Japan
Since dental pulp tissue with irreversible pulpitis lacks self-regenarative ability, it is usuallycompletely removed and replaced by artificial materials. The teeth that received pulpectomy lose their
physiological bioactivity, including strength, sensitivity and immune defense, and many cases
ultimately require extraction because of fractures or secondary caries. Therefore reconstruction ofdentin/pulp-complex is considered as an ideal treatment for pulpectomized teeth. Autologoustransplantation of human dental pulp stem cells (DPSCs) is promising treatment to reconstructdentin/pulp-complex because of their capacity to regenerate dentin. Unfortunately, it is very limited
opportunity to isolate healthy DPSCs from patients. Therefore, irreversible pulpitis-derived dental pulp
stem cells (IP-DPSCs) are considered to be useful resource for autologous DPSCs based dentin/pulp
regenerative therapy. We succeeded in isolating IP-DPSCs from ectomized dental pulp tissues which
was diagnosed as irreversible pulpitis. IP-DPSCs expressed lower stem cell properties,
immunosuppressive functions and dentin/pulp regenerative capacity in comparison with healthy
DPSCs.
In this study we attempted to improve these failure functions of IP-DPSCs to enabledentin/pulp-complex regeneration. In vitro pre-treatment with interferon-gamma (IFN-$) improved invivo dentin/pulp regeneration of IP-DPSCs when they were implanted with hydroxyapatite/tricalcium
phosphate carrier under the dorsal skin of immunocompromised mice. Moreover, pre-treatment with
IFN-$ improved immunosuppressive function of IP-DPSCs. To confirm whether IP-DPSCs can beapplied dentin/pulp regenerative therapy, we transplanted in vitro IFN-$ pre-treated IP-DPSCs into root
canal of human extracted teeth and transplanted them with hydroxyapatite/tricalcium phosphate carrier
under the dorsal skin of immunocompromised mice. After 8 weeks, transplanted tissues showeddentin/pulp-complex like structure. IFN-$ pre-treated IP-DPSCs expressed enhanced capacity to form
newly dentin like structure on the surface of human dentin.
These findings suggest that IFN-$ pre-treatment is effective method to improve impaired function of
IP-DPSCs. Moreover, IP-DPSCs are promising resource for DPSCs based dentin/pulp-complex
regenerative therapy.
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