Kyudai Oral Bioscience 2018
(KOB2018)
Health Longevity from Oral Brain Science:
From Basic to Clinical Research
February 11th (Sun) - 12th (Mon) 2018
Collaboration Station I, 2F Audiovisual Hall
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 aUSB. 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 to the 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.

2

PROGRAM3

Kyudai Oral Bioscience 2018 (KOB2018)
Feb 11 (Sun) 13:0017:10
13:0013:10
Hiroshi Nakanishi(Representative Organizer of KOB, Vice Dean)

Opening Remarks
Kiyoshi Koyano(Dean, Faculty of Dental Science, Kyushu University)

Symposium
Healthy Longevity from Oral Brain Science:
From Basic to Clinical Research
Session 1

Oral Health and Cognitive Function

Chair person Noriatsu Shigemura (Kyushu University)
13:1013:40 Special Lecture 1
Tetsuya Goto (Kagoshima University)
Neurodegeneration by Tooth Loss Causes Cognitive Impairment
13:4014:10 Special Lecture 2
Makoto Inoue (Niigata University)
Physiological Study on Swallowing Performance
14:1014:20

Session 2

Coffee Break

Periodontitis and Alzheimer’s Disease

Chair person Hiroshi Nakanishi(Kyushu University)

14:2015:10 Special Lecture 3
Angela R Kamer (New York University)
Peripheral Inflammatory Conditions Contribute to Alzheimer's Disease:
the Periodontal disease model4

15:1015:40 Special Lecture 4
Zhou Wu (Kyushu University)
Mechanisms Linking Periodontal Disease to Alzheimer's Disease: Key functionsof cathepsins in microglia activation and chronic systemic inflammation
15:4015:50

Session 3

Coffee BreakTreatments and Biomarkers for and Alzheimer’s Disease

Chair person Haruhiko Kashiwazaki(Kyushu University)

15:5016:20 Special Lecture 5
Hachiro Sugimoto (Doshisha University)
Development of Drug (GT863) for Alzheimer’s Disease
16:2016:50 Special Lecture 6
Kazuhiko Uchida (Tsukuba University)
Sequester Proteins Involved in Amyloid-β Clearance as Blood-based Biomarkers
for Alzheimer’s Disease
16:5017:00

Group Photograph
17:0017:10

Closing Remarks
Fusanori Nishimura(Vice Dean, Faculty of Dental Science, Kyushu University)5Feb 12 (Mon) 9:3012:35

9:309:40

Opening Remarks
Ichiro Takahashi(Vice Dean, Faculty of Dental Science, Kyushu University)

Educational Lectures
Chair person Ichiro Takahashi(Kyushu University)

9:4010:10 Educational Lecture 1
Haruhiko Kashiwazaki (Kyushu University)
Oral Supportive Care for Cancer Therapy
Chair person Kasuaki Nonaka
10:1011:00 Educational Lecture 2
Janice A Townsend (Louisiana State University)
Innovation in Education and Research at LSU School of Dentistry
11:0011:10

Coffee BreakPhD Student Session (PSS)
Chair personsYicong Liu & Yuka Harada
11:1011:25 PSS-1
Eddy (Departments of Biometerials)
Fabrication of self-setting β-tricalcium phosphate granular cement throughbridging with calcium sulfate hemihydrate
11:2511:40 PSS-2
Yuka Harada (Department of Aging Science and Pharmacology)
Cathepsin E-dependent elastase secretion in neutrophils derives experimental autoimmune
encephalomyelitis-induced neuropathic pain6

11:4011:55 PSS-3
Misaki Abe (Laboratory of Oral Pathology of Dental &Section of Oral and Maxillofacial
Surgery)

Expression and function of Piezo1 and Piezo2 in tooth germ development
11:5512:10 PSS-4
Mai Arima (Department of Endodontology and Operative Dentistry)
R-spondin2 enhances osteogenesis of immature human periodontal ligament
cells through the canonical Wnt signaling pathway
12:1012:25 PSS-5
Keita Funada (Section of Orthodontics and Dentofacial Orthopedics)
The roles of miR875-5p as a specific marker during early tooth development

Closing Remarks
12:2512:35
Seiji Nakamura(Associate Director of Kyushu University Hospital)

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ABSTRACTS

8

Symposium
Health Longevity from Oral Brain Science:
From Basic to Clinical Research9Session 1
Oral Health and Cognitive Function

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Special Lecture 1
Neurodegeneration by Tooth Loss Causes
Cognitive Impairment
Tetsuya Goto
Division of Oral Anatomy and Cell Biology, Graduate School of
Medical and Dental Sciences, Kagoshima University,
Kagoshima 890-8544, Japan

Brief curriculum vitae: Dr. Goto graduated from Kyushu University in 1988, completed Doctoral
Course of Kyushu University and received a PhD degree in Dental Science in 1992. Served as a
post-doctoral fellow at University of British Columbia and University of Toronto in Canada until
1996. Worked as Assistant Professor at Kyushu University, until 2001, and as Associate Professor
at Kyushu Dental University until 2014. Currently, working as a Professor at Faculty of Dentistry,
Kagoshima University.

Abstract: Alzheimer’s disease (AD) is a major cause of dementia in elderly people. The mainneuropathological changes of AD are amyloid beta (Aβ) deposition, phosphorylation of tau (pTau),and neural cell loss in the brain. Poor oral health condition such as periodontitis or tooth loss has
been thought to be involved in the progression of AD. However, it was unclear how
neurodegeneration by tooth loss affects the progress of AD. At first, we confirmed which trigeminal
neurons was induced the neurodegeneration of AD in 3xTg AD model mice using
immunohistochemistry. Then, we found the strong Aβ deposition and pTau in the neuron at thetrigeminal mesencephalic nucleus (Vmes) among trigeminal nuclei. Next, we extracted bilateralmaxillary molars of 12-month-old 3×Tg AD model mice and examined the pathological changes.
At one-month after tooth extraction the number of Aβ- and pTau-immunoreactive neurons at thecaudal side of Vmes was intensively reduced. Iba-1 immunoreactive microglia distributed aroundthe neural loss site of Vmes. Close to the caudal site of Vmes with microglia, there islocus coeruleus (LC) that is well know the place where the initial lesion of AD appears and its cell
death causes a reduction in nerve cells in the hippocampus. We also found the neural loss of LC
neuron with active microglia close to the place where neural loss of Vmes was occurred after tooth11

extraction. Therefore, we propose the following cascade model based on these results. After the
extraction of the molars in AD model mice, neurons on the caudal side of Vmes cause cell death.
Dead neurons are phagocytosed by microglia, coinstantaneously Aβ is released and activates
microglia. Activated microglia also phagocytose the neighboring neurons of LC. Deposition of
Aβ on neurons is known to begin at the early age, indicating that tooth loss is a risk factor of AD.

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Special Lecture 2
Physiological study on swallowing performance
Makoto Inoue
Division of Dysphagia Rehabilitation, Niigata University Graduate
School of Medical and Dental Sciences, Niigata, Japan

Brief curriculum vitae: Dr. Inoue graduated from Niigata University in 1994, completed Doctoral
Course of Niigata University and received a PhD degree in Dental Science in 1998. Worked as
Assistant Professor at 1998, until 2004. Worked as Lecturer at 2004, until 2006. Worked as
Associate Professor at 2006, until 2008. Currently, working as a Professor at 2008, Niigata
University.

Abstract:

Japanese population is aging, and more than 27% to total population is now over 65

years old. One of the most common cause of death in Japanese elderly people is aspiration
pneumonia caused by dysphagia. Dysphagia is a symptom of difficulty of swallowing due todiseases such as stroke, neurological diseases, head and neck cancer and so on. Major pathologicalproblems of dysphagia is decline of swallowing initiation. In my presentation, I will introduce ourbasic study to investigate which type of receptors in the pharyngeal/laryngeal cavity is involved inswallowing initiation. To initiate swallow, thermal, chemical or electrical stimulation is available.
Thermal and chemical stimulation may be expected to activate some types of channels, i.g. TRPs,
ASICs etc. On the other hand, electrical stimulation is used to widely activate the peripheral nerve
or nerve endings. Peripheral stimulation can activate swallow-related neural network not only in thebrain stem but also in the higher centers such as the cerebral cortex. We will also introduce ourhuman study to investigate how we evaluate the swallow-related neural activity.

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Memo

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Session 2
Periodontitis and Alzheimer’s disease16

Special Lecture3
Peripheral Inflammatory Conditions Contributeto Alzheimer's Disease:
the Periodontal disease modelAngela Kamer
Department of Periodontology and Implant Dentistry,
New York University (NYU) College of Dentistry, New York, USA.

Brief curriculum vitae: Dr. Kamer is associate professor in the Department of Periodontology
and Implant Dentistry, NYU College of Dentistry, New York, USA. Her research activities
involved periodontal disease, implant dentistry, diabetes, cancer, bone research and currently
Alzheimer’s disease. After completing her studies in Stomatology at the University "Iuliu
Hatieganu" in Cluj Napoca, Romania, and practicing dentistry for a few years, Angela Kamer went
on to receive her MS in Oral Sciences, Ph.D. in Pathology and training in Periodontics from SUNY
at Buffalo and later in Implant Dentistry from NYU, USA. Subsequently, she started working at
NYU College of Dentistry. As a trained periodontist at the NYU College of Dentistry and a
research scientist at NYU Center for Brain Health, Angela is in a unique position to bridge themedical-dental research gap by developing new hypotheses and strengthening collaborations acrossdisciplines. Her research focuses on the role of peripheral inflammatory and dysbiotic conditions in
the pathogenesis of Alzheimer’s disease using periodontal disease as a model. Periodontal disease isa peripheral chronic polymicrobial inflammatory condition affecting about 50% of people over 55
years of age in USA. It is characterized by high Gram-negative pathogenic bacterial load and often
increased systemic inflammation. Studies coming from Angela’s group will present data showingthat periodontal disease may contribute to AD-specific pathology and cognitive performance. She
published in well-regarded peer-reviewed journals, she wrote book chapters, and presented toseveral international conferences. She also co-edited a book entitled "A Clinician's Guide to
Systemic Effects of Periodontal Diseases" and a special journal issue.
Abstract:

Worldwide, more than 31 million persons suffer from dementia, with 50-60% having

AD and these numbers are estimated to double by 2030 and double again by 2050. Alzheimer's
disease is a continuum condition characterized by specific pathology such as amyloid β plaques(Aβ), neurofibrillary tangles, inflammation and neurodegeneration. In the preclinical phase, there is17

AD pathology but no symptomatology suggesting that the AD pathology did not cause irreversible
brain damage. Therefore, identifying potentially reversible risk factors before or during the
preclinical phase would provide an opportunity for prevention and intervention.
It is accepted that Inflammation is involved in the pathogenesis of AD, but it remains unknown
to what extent peripheral inflammation and infections and dysbiotic conditions play a role. Studies
have shown that peripheral cytokines, infections and their products including bacterially derived
lipopolysaccharide (LPS) can promote AD-specific pathology. Periodontal disease a localized
peripheral chronic inflammatory condition affecting about 50% of people over 55 years of age ischaracterized by dysbiosis with high Gram-negative pathogenic bacterial load and increased
systemic inflammation.
It is our general hypothesis that peripheral sources of inflammation, with its
inflammatory/bacterial burden, contribute to Alzheimer’s disease (AD) pathogenesis by altering
AD-specific pathology. In support of this hypothesis we find that in cognitively normal individuals
clinical measures of periodontal disease associate with brain amyloid load (assessed by 11C-PIB
PET imaging) in AD amyloid-vulnerable regions and enhances tau pathology thus, supporting a roleof peripheral inflammations/infections in AD pathogenesis.

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Memo

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Special Lecture 4
Mechanisms Linking Periodontal Disease to
Alzheimer's Disease:
Key functions of cathepsins in microglia
activation and chronic systemic inflammation
Zhou Wu
Department of Aging Science and Pharmacology, Faculty of
Dental Science, Kyushu University, Fukuoka 812-8582, Japan
Brief curriculum vitae: Dr. Wu graduated from Bethune Medical College of Ji Lin University in
China, completed Doctoral Course of Kyushu University and received a PhD degree in Dental
Science. Served as a research fellow of Japan Society for the Promotion of Science at Kyushu
University from 2002 to 2004. Worked as Assistant Professor, Lecturer and Associate professor at
Kyushu University Faculty of Dental Sciences from 2005 until now. She is a visiting Professor at Ji
Lin University and China Medical University, China. She is also a Diamond Jubilee International
Visiting Fellow in University of Southampton, UK. Her research interest is functions of cathepsins
in innate and adaptive immunity that regulate functions of mononuclear phagocytes including
dendritic cells and microglia during chronic systemic inflammation.
Abstract: Many lines of evidence show that chronic neuroinflmmation play an essential role inonset and progression of Alzheimer's disease (AD). Furthermore, there is increasing evidence thatchronic systemic inflammation enhances neuroinflammation through activation of microglia.
During the last decade, numerous clinical studies have found that as a chronic oral infectious
disease, periodontal disease, is correlated with AD and is involved in persisting systemic
inflammation. We have previously found that cathepsin B, a lysosomal cysteine protease, isinvolved in microglia-mediated neuroinflammation, which induces memory impairment. Therefore,
we have proposed "microglia-aging" hypothesis, which microglia-mediated neuroinflammation
promotes the brain aging [1]. Lipopolysaccharide derived from Porphyomonas gingivalis (Pg-LPS),
a major causative agent of periodontal disease, induces microglia-mediated neuroinflammation
through activating leptomeningeal cells, which cover the entire cortex [2]. Furthermore, the chronicsystemic exposure of Pg-LPS in low concentration causes the cathepsin B-dependent AD-like
pathologies, including microglia-mediated neuroinflammation, the amyloid β accumulation in20

neurons and memory impairment in middle-aged mice [3]. More recently, we have shown thatchronic systemic exposure of Pg-LPS amplifies cathepsin S-dependent systemic inflammation bypromoting the differentiation of Th17 cells through activation of dendritic cells in middle-aged mice
[4].
These findings demonstrate that cathepsins B and S are key enzymes for controlling
microglia-mediated neuroinflmmation and the systemic inflammatory signaling from periodontal
disease to the brain. Because AD is a disease that progresses with a long span of more than 25 years
and fundamental therapeutic agents have not yet been developed, it is important to prevent or delay
the onset and progression of AD. As a treatable disease, oral care for periodontal disease will reduce
the risk for AD. In addition to the dental treatment, specific inhibitors of cathepsin B or S can beused in the prevention and treatment of AD.

References
1. Nakanishi H, Wu Z. Microglia-aging: roles of microglial lysosome- and mitochondria- derived
reactive oxygen species in brain aging. Behav Brain Res. 201(1), 1-7, 2009.
2.

Liu Y, Wu Z, Zhang X, Ni J, Yu W, Zhou Y, Nakanishi H. Leptomeningeal cells transduce
peripheral macrophages inflammatory signal to microglia in response to Porphyromonas
gingivalis LPS. Mediators Inflamm 2013:407562, 2013.

3.

Wu Z, Ni J, Liu Y, Teeling JL, Takayama F, Collcutt A, Ibbett P, Nakanishi H. Cathepsin B
plays a critical role in inducing Alzheimer’s disease-like phenotypes following chronicsystemic exposure to lipopolysaccharide from Porphyromonas gingivalis in mice. Behav Brain
Immun 65: 350-361 2017.

4.

Dekita M, Wu Z, Ni J, Zhang X, Liu Y, Yan X, Nakanishi H, Takahashi I. Cathepsin S is
involved in Th17 differentiation through the upregulation of IL-6 by activating PAR-2 after
systemic exposure to lipopolysaccharide from Porphyromonas gingivalis. Front Pharmacol
8:470, 2017.

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Memo

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Session 3
Treatments and Biomarkers for
Alzheimer’s disease23

Special Lecture 5
Development of Drug (GT863) for
Alzheimer’s Disease
Hachiro Sugimoto
Doshisha University Faculty of Life and Medical Science,
Kyoto, Japan.

Brief curriculum vitae: Dr. Sugimoto joined Eisai Co., Ltd. in 1961. While working for Eisai, he
graduated from Chuo University in 1969 and obtained his doctorate in pharmacology at Hiroshima
University in 2002. In 2003, he resigned from Eisai, and became Professor in the Graduate Schoolof Pharmaceutical Sciences, Kyoto University. Currently, working as a Professor at Doshisha
University Faculty of Life and Medical Science.
His research on E2020 (now called Donepezil), an acetylcholinesterase inhibitor, first began at
Eisai's Tsukuba Research Laboratories in 1983, because his mother suffered from dementia. His
research group was finally able to successfully create Donepezil with a promising enough profile
for the compound to become a drug candidate. For his achievement, he received the Award of
Science from Eisai in October 1993, the Award of Engineering from the Pharmaceutical Society ofJapan in March 1998, the Special Galien Prize in the UK in April 1998, the Chemistry-Bio Tsukuba
Award in May 1998, and the Imperial Award for Invention in June 2002.
Abstract:
Outline of disease
Alzheimer’s disease (AD) starts with an early memory impairment, gradually decreases
comprehension ability and judgmental ability, interferes with social life, and then nursing care isrequired in 5 to 6 years. Eventually it is also called "a disease that loses personality" and it is one of
the most cruel diseases among all ones. In Japan, about 4 million people suffer from dementia, and4 million people are said

mild cognitive impairment (MCI) and the number of patients is rapidly

increasing (Ministry of HealthLabor and Welfare in Japan). About 2/3 of dementia patients are
Alzheimer's disease, the incidence of disease has increased nearly threefold since 1990 and it isexpected to increase to 4 times from now in 2050.

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Therapeutic drug development
The biggest challenge for AD is the lack of effective radical therapies. Until now, thedevelopment of a large number of therapeutic drugs has been promoted by pharmaceuticalcompanies, universities, laboratories and venture companies, but everything has failed at the clinical
trials. GT 863 is a candidate drug for AD with new profiles developed by Green Tech Co., Ltd., andis promoting commercialization ahead of the rest of the world.

Current status of development of therapeutic drug for AD
Development based on amyloid hypothesis: Target the inhibition of Aβ cascade. Amyloid β(Aβ) has been considered the most noteworthy as a therapeutic target. Aβ is elevated in familial AD
patients, and APP as a precursor is also elevated in Down syndrome patients who become youngAD, so it has been noted as a potent inducer of AD. Furthermore, aggregates of Aβ actually haveneurotoxicity (inducing nerve cell death).
Development of therapeutic drugs
1) The development of enzyme inhibitors of β / γ secretase responsible for the production ofAβ from APP was advanced. However, the development has been halted because of strong
side effects, no efficacy, or symptomatic worsening.
2) Development of drugs targeting inhibition of the formation of aggregates of Aβ has been
advanced, and the development of small molecules and antibodies, but everything has failedin phase 3 clinical trials.

Development based on the Tau hypothesis : Target the inhibition of tau cascade factor Tau.
Factors that have recently drawn attention as therapeutic targets are neurofibrillary tangles
accumulated in neurons, an aggregate of phosphorylated tau protein. It has been confirmed that thisaggregate possesses neuro cytotoxicity and has attracted attention as a therapeutic target for AD.
However, the development of AD therapeutic drugs targeting tau has only just begun, there are few
examples that have been developed yet. Currently, all clinical trials, including the first compoundMB, have failed.

AD Patient: Onset mechanism of disease
Presently, hundreds of AD therapeutic drugs are probably developed in the world and clinical25

trials are being conducted, but all of them have failed to develop. Most of these developments target
Aβ or Tau, but unlike these developments, when developing new AD therapeutic drugs in the future,
it is necessary to construct an accurate development strategy based on an accurate understanding ofAD pathology.
Accurate understanding of AD pathology

AD is caused by "neuronal cell death"
AD is a disease leading to a significant loss of reduced cognitive brain function by celldeath

in the brain. Neurons decrease due to cell death and the brain atrophies. → Detected as remarkable
brain atrophy image of MRI.

Neuronal cell death is induced by "Aβ" and "Tau"
Characteristic pathological image of AD patients   Amyloid β (Aβ) aggregates accumulate outside the brain cells (senile plaques)    Aggregates of phosphorylated tau proteins appear in cells
Since neurofibrillary tangles directly induce nerve cell death, it is also a potential therapeutictarget. Aβ and Tau are both promoted simultaneously in the majority of AD
patient tissues. Both are expressed simultaneously in the same patient.

Development strategy for AD therapeutic drug (GT863)
Compound synthesis was conducted with the aim of the concept of "compounds thatsimultaneously inhibit both Aβ aggregation and Tau aggregation", which is considered to be an
essential requirement for AD therapeutic drug development. Curcumin has an inhibitory effect on
aggregation of Aβ · Tau at the in vitro / in vivo experimental level. However, it did not show
reasonable efficacy in clinical trials for AD. The reason for this is that curcumin itself is still low
aggregation inhibitory activity, bad pharmacokinetics (Low absorptivity, easy metabolism, lowbrain migration). We have synthesized approximately 1,000 compounds using this curcumin as aseed compound and searched for compounds with high inhibitory activity on aggregation of
Aβ · Tau and high pharmacokinetics. As a result, GT863 was obtained as the most suitablecompound.

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Memo

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Special Lecture 6
Sequester Proteins Involved in Amyloid-β
Clearance as Blood-based Biomarkers for
Alzheimer’s Disease
Kazuhiko Uchida
Department of Molecular Biology and Oncology,
Faculty of Medicine, University of Tsukuba, Tsukuba , Japan
Brief curriculum vitae: Dr. Uchida graduated from Nara Medical University in 1983, completed
Doctoral Course of Nara Medical University and received a M.D., Ph.D. degree in Medical
Sciences in 1988. Served as a Research Resident at National Cancer Center Research Institute from1985 to 1989. Worked as Research Staff at National Cancer Center Research Institute in 1989.
Worked as Assistant Professor at University of Tsukuba, until 1995. Currently, working as an
Associate Professor and Research Staff at Center for Tsukuba Advanced Research Alliance, atUniversity of Tsukuba, and CEO/CTO at MCBI, Inc., a spin-out company from University ofTsukuba.

Abstract: Currently, the number of people who have been diagnosed with dementia worldwide is35.6 million, and it is predicted that this rate will increase to more than 100 million people in 2050.
Diagnosis and intervention at early stages of dementia may greatly reduce the number of individuals
suffering from this debilitating disease. Mild cognitive impairment (MCI) is a syndrome defined ascognitive decline that is greater than expected for an individual's age and is regarded as a risk groupfor dementia. MCI due to Alzheimer’s Disease (AD) is considered as a pre-stage of AD, andwithout clinical intervention, 40% of patients with MCI may convert to AD within four years ofdiagnosis.
Biomarker discovery for neurological disorders is challenging but rewarding because it
facilitates early diagnosis, monitors disease progression, and assesses response to medical treatment.
Development of blood-based biomarkers for early detection of cognitive impairment is important topotentially reduce the number of patients with dementia.
AD is distinguished from other neurological diseases, including other types of dementia andnormal aging, by abundant deposits of fibrillar amyloid β (Αβ). Aβ-derived diffusible ligands cause
28abnormal spine morphology and a significant decrease in spine density, and trigger synaptic
dysfunction, which may lead to memory loss in AD. In healthy individuals, synapse damage by
amyloid-β (Αβ) is prevented by clearance mechanisms via blood-brain-barrier. Apolipoprotein andtransthyretin are involved in sequestration of Αβ from brain to blood. Aβ may also be removed byclearance mechanisms such as phagocytosis by microglia. Complement proteins sequestrate Αβ via
an innate immune response of microglia in the brain.
We analyzed serum levels of apolipoprotein A1 (apoA1), complement protein C3 (C3), and TTR
in development of blood-based biomarkers for MCI and AD, and revealed the clinical potential ofthese proteins involved in sequestration of Αβ in the assessment of cognitive decline by longitudinaland cross-sectional studies using 3 independent cohorts1. The least absolute shrinkage and selection
operator (LASSO) was used to evaluate the combination of multiple biomarkers in MCI vs. NDC.
A combination of apoA1, C3, and TTR achieved an area under the curve of 0.89 in MCI vs. healthy
controls and also discriminated individuals with mild cognitive decline from non-demented
age-matched control (NDC).
We further developed conventional immuno-assays for these sequester proteins to investigate
their clinical validity as blood-based test for MCI. In addition, we developed novel C3 assay which
differentially measured a native form (nC3) and activated (fragmented) forms (aC3) of C3
indicating status of activation of C3. The levels of theses serum biomarkers were compared with
MMSE score, voxel-based specific regional analysis system for Alzheimer Disease (VSRAD) scorein MRI and reginal cerebral blood flow (rCBF) in SPECT from 257 participants.
A combination apoA1, TTR and aC3/nC3 achieved an area under the curve of 0.89 in MCI vs.
NDC. Atrophy in medial temporal were associated with decreased levels of apoA1 and HDL.
Significantly lower levels of aC3, apoA1, HDL, and total cholesterol were observed in participants
with severe reductions in rCBF. Furthermore, lower nC3 and higher aC3 levels were observed in
AD hippocampus tissues by immunohistochemical analysis.
The current results are consistent with the hypothesis that individuals with decreased levels or
impaired function of sequester proteins resulting in impaired Αβ clearance are susceptible tocognitive impairment. Dysfunction of sequestration and/or low levels of sequester proteins may
promote pathological progression of AD. Thus, apoA1, TTR and complement protein profiles could
be blood-based biomarkers for assessment of early stages of cognitive decline including of
preclinical AD.

29

1. Uchida, K., et al. Amyloid-β sequester proteins as blood-based biomarkers of cognitive decline.
Alzheimers & Dement (Amst), 1, 270-280 (2015).

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Educational Lectures

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Educational Lecture 1
Oral Supportive Care for Cancer Therapy
Haruhiko Kashiwazaki
Division of Maxillofacial Diagnostic and Surgical Sciences, Facultyof Dental Science, Kyushu University, Fukuoka 812-8582, Japan

Brief curriculum vitae: Dr. Kashiwazai graduated from Hokkaido University in 1992, completed
Doctoral Course of Hokkaido University and received a PhD degree in Dental Science in 1997.
Served as a post-doctoral fellow at Hokkaido University until 2001. Worked as Assistant Professor
until 2013, as Lecturer until 2015 at Hokkaido University. Currently, working as a Professor at
Section of Geriatric Dentistry and Perioperative Medicine in Dentistry, Kyushu University.
Abstract:
Oral supportive care is required as part of the cancer therapy pretreatment assessment tomanage conditions that may impact the cancer therapy and affect the ability of the person tocomplete the planned therapy. Programs to prevent acute and chronic complications of the cancer
treatment are initiated.
Oral complications of cancer therapy can be the most distressing of side effects and affectquality of life.

The acute complications include oral infection, oral mucositis, altered saliva,change in taste, and coincidental dental infection, all of which can result in pain, and affect all
aspects of oral function.
Prevention and management of chronic complications and observance for recurrent or second
primary cancers is mandatory following cancer therapy. The most distressing chronic complication
is dry mouth, which leads to increased risk of dental cavities, gum disease, mucosal infection,
reduced taste, dysphagia, poor denture function and affect nutrition.
Oral supportive care providers, acting as part of the medical team, can facilitate the delivery ofcancer treatment, and affect the quality of life during and following cancer therapy.

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   Educational Lecture 2
Innovation in Education and Research at
LSU School of Dentistry
Janice A. Townsend
Department of Pediatric Dentistry at LSU School of Dentistry in
New Orleans, Louisiana, USA.

Brief curriculum vitae: Dr. Townsend is Associate Professor and Chair of the Department ofPediatric Dentistry at LSU School of Dentistry in New Orleans, Louisiana. She received a bachelorof arts in English with a minor in chemistry in 2001 from the University of North Carolina (UNC)
in Chapel Hill, received her doctorate of dental surgery from Marquette University School of
Dentistry in Milwaukee, Wisconsin, in 2005 and then completed her certificate in pediatric dentistry
and her master of dental science at the Ohio State University in Columbus, Ohio, in 2007. In 2008,
she received her board certification from the American Board of Pediatric Dentistry and in 2009,
awarded the Art Nowak Award for scoring the highest on the board’s oral clinical examination. She
has served on numerous committees for the university, including the leadership committee for the
LSUHSC accreditation process for Southern Association of Schools and Colleges. She also has
served on Children’s Hospital of New Orleans Medical Executive Committee. Her current areas ofresearch and publication are pain control in children, behavior guidance dental education, andbiomaterials.
Abstract: LSU Department of Pediatric Dentistry is a comprehensive unit that provides dental
education to pediatric dental residents, dental students, and dental hygiene student, conducts
research, and provides service to the community in New Orleans Louisiana.

The department isenhancing the educational experience of pediatric dental residents and dental students throughutilization of a pioneering “flipped classroom” education design and objective structured clinicalexamination (OSCE) assessment.

This talk will address how both of these pedagogicalinnovations can be implemented into any department.
LSU Department of Pediatric Dentistry is also a leader in the area of biomaterials research,especially in the area of sealants.

Recent department research is addressing fundamental gaps inknowledge pertaining to sealant placement techniques as well as partnering with well-known basic
science researchers to revolutionize the sealant composition to better inhibit caries.
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36

PhD Student Session

37

PSS-1Fabrication of self-setting β-tricalcium phosphate granular cement
through bridging with calcium sulfate hemihydrate
Eddy1, Akira Tsuchiya1, Kanji Tsuru2, Kunio Ishikawa1
1

Department of Biomaterials, Faculty of Dental Science, Kyushu University, Fukuoka 812-8582,

Japan, 2Section of Bioengineering, Department of Dental Engineering, Fukuoka Dental College,
2-15-1 Tamura, Sawara-ku, Fukuoka 814-0193, Japan.
Introduction: Previously, we have proposed beta tricalcium phosphate granular cement(β-TCPGC) that is useful to prevent flowing out of the β-TCP granules from the bone defect. Whenthe β-TCP granules were mixed with acidic calcium phosphate solution, it set to form
interconnected porous structure. Although it seems promising, β-TCP granules need to be mixed
with the acidic calcium phosphate solution. Calcium sulfate hemihydrate (CSH) has self-setting
ability by converting to calcium sulfate dihydrate (CSD) when exposes to water. Furthermore, CSD
is the component of Osteoset®, which is a commercially available as a bone substitute. In this study,
we fabricated CSH coated β-TCP granules through dissolution-precipitation method. This setting
ability could inhibit the flowing out of the β-TCP granules from the bone defect.
Experiment method: β-TCP granules were immersed in NaHSO4 solution for 1, 3, 5 and 7 days
at 70oC. The samples were then heated at 120oC for 4 hours. β-TCPGC was prepared by mixing the
granules with saline solution at a L/P ratio of 0.3 and identified by XRD and SEM. The mechanicalstrength of the β-TCPGC was measured as a DTS by universal testing machine. β-TCPGC
implanted in rabbit femur for 4 weeks and the percentage of newly formed bone was calculated
from histological analysis.
Results: β-TCP granules immersed in NaHSO4 solution were coated by CSD and CSD became
CSH after heating at 120ºC for 4 hours. CSH coated β-TCP granules mixed with saline solution
were set and DTS value of β-TCPGC with 75 wt% of CSH was 0.8±0.1 MPa. The percentage of
newly formed bone of β-TCPGC with 75 wt% of CSH was 28.7±0.5 %, meanwhile β-TCP granules
without coating was 19.9±1.1 %.
Conclusion: CSH coated β-TCP were successfully fabricated and formed interconnected porous
structure with good mechanical strength after mixed with saline solution.
Key words: beta tricalcium phosphate, interconnected porous structure, calcium sulfate

38

PSS-2Cathepsin E-dependent elastase secretion in neutrophils derives
experimental autoimmune encephalomyelitis-induced neuropathic
pain
Yuka Harada, Yoshinori Hayashi, Zhou Wu, Hiroshi Nakanishi
Department of Aging Science and Pharmacology, Kyushu University,
Fukuoka 812-8582, Japan
Multiple sclerosis (MS) is a chronic disease characterized by the destruction of central nervous
system (CNS). Infiltration of immune cells into CNS is a hallmark of MS, leading to neuronal
dysfunction such as motor disorder and paresthesia. It has been estimated that 55 % of MS patients
experienced pain symptom that cause sleep disturbance, leading to fatigue and depression, andseriously affect the patient's daily activities and quality of life. Although pain is a common problem
in MS patients, no effective therapeutics are commercially available. In the present study, we try tofind molecular mechanism of neuropathic pain in MS using by experimental autoimmune
encephalomyelitis (EAE), a mouse model of MS.
Neuropathic pain in C57BL/6 mice was observed 3 days after myelin oligo-glycoprotein(MOG) immunization before EAE onset (days 9-12 post immunization). In contrast, cathepsin E(CatE)-knockout mice were highly resistant to neuropathic pain caused by MOG-immunization. We
observed that neutrophils infiltrated into the dorsal root ganglion (DRG) after MOG immunization.
Isolated neutrophils from bone marrow increased both protein and mRNA levels of CatE following
MOG stimulation. Adoptive transfer of WT neutrophils that stimulated with MOG induced pain
behaviors in recipient mice, whereas that of CatE-/- neutrophils did not. Further study revealed thatsivelestat, a neutrophil elastase specific inhibitor, significantly suppressed neuropathic pain causedby adoptive transfer of MOG-stimulated neutrophils. In addition, neutrophils increase elastaseactivity in response to MOG stimulation, which was a CatE-dependent manner. Elastase derived
from neutrophils activated protease-activated receptor-2 (PAR2) in the DRG neurons, which
triggered neuropathic pain in EAE. We finally identified that CatE induction in neutrophils by
MOG was mediated by TLR4.
Together, CatE in neutrophils thus become a promising therapeutic target for neuropathic pain
in MS patient.
Key words: multiple sclerosis, EAE, neutrophils, Cathepsin E, neutrophil elastase39

PSS-3
Expression and function of Piezo1 and Piezo2
in tooth germ development
Misaki Abe 1,2, Hiroko Wada 1, Tamotsu Kiyoshima 1
1
2

Laboratory of Oral Pathology, of Dental, Kyushu University, Fukuoka, 812-8582, Japan

Section of Oral and Maxillofacial Surgery, Division of Maxillofacial Diagnostic and Surgical
Sciences, Faculty of Dental, Kyushu University, Fukuoka, 812-8582, Japan,

Mechanotransduction, the conversion of mechanical force into biological signals, has a crucialrole in physiology. In mammals, embryonic development, touch, pain, adjustment of vascular tone
and blood flow, lung growth, and bone and muscle homeostasis are all regulated bymechanotransduction (Coste B et al., Science, 2010). The epithelial-mesenchymal interaction in thetooth germ development dynamically produces to figure complicated tooth forms, however it isunknown whether Piezo family is involved as mechanosensitive receptor in the developing tooth
germ. In this study, we examined to clarify expression patterns and functions of Piezo1 and Piezo2
in tooth germ development.
At the bud and cap stages in the mouse lower first molar, Piezo1 mRNA expression wasprominently observed in the enamel organ, while Piezo2 mRNA expression was seen in the enamelorgan and the surrounding mesenchymal cells. At the bell stage, Piezo1 and Piezo2 mRNA
expression was detected in both the ameloblasts and odontoblasts. At the tooth root formation stage,
Piezo1 and Piezo2 mRNAs were noted in the odontoblasts of the tooth root. The expression patternsof the Piezo1 and Piezo2 proteins were similar to those of the Piezo1 and Piezo2 mRNAs,
respectively. In addition, to investigate the effects of knockdown of Piezo1 on the tooth germmorphogenesis, organ culture was performed in cultured embryonic day 15.0 (E15.0) for 8 days.
Tooth germs treated with Piezo1 siRNA showed irregular arrangements of the ameloblasts andodontoblasts. Immunochemical staining for Ki-67 was carried out to examine the cell proliferative
activity in the tooth germs since the development of the E15.0 tooth germs was disturbed bytreatment with Piezo1 siRNA. The number of Ki-67 positive cells increased in theirregular-arrangemented ameloblasts in the tooth germs treated with Piezo1 siRNA.
These results suggested that Piezo1 and Piezo2 might play functional roles in tooth germdevelopment.
Key words: Piezo1, Piezo2, mechanosensitive channels
40

PSS-4R-spondin2 enhances osteogenesis of immature human periodontal
ligament cells through the canonical Wnt signaling pathway
Mai Arima1, Daigaku Hasegawa2, Shinichiro Yoshida2, Hiromi Mitarai2,
Atsushi Tomokiyo2, Sayuri Hamano1,4, Hideki Sugii2,
Naohisa Wada3, Hidefumi Maeda1,2
1

Department of Endodontology and Operative Dentistry, Faculty of Dental Science, Kyushu
University, 2Division of Endodontology, Kyushu University Hospital, 3Division of General
Dentistry, Kyushu University Hospital, 4OBT Research Center, Faculty of Dental Science, Kyushu
University, Fukuoka 812-8582, Japan
R-spondin family proteins (RSPOs), secreted glycoproteins, are known to play an importantrole in embryonic development and tissue homeostasis through activation of the canonicalWnt/β-catenin signaling pathway. R-spondin2 (RSPO2), a member of RSPOs, has been reported toenhance osteogenesis in mice. In this study, we examined the localization of RSPO2 in periodontal
ligament (PDL) tissue and the effect of RSPO2 on osteoblastic differentiation of PDL cells.
Immunohistochemical staining revealed that RSPO2&was&expressed&in&rat&PDL&tissue.
Semi-quantitative RT-PCR and immunofluorescence staining demonstrated that&the&immature
human PDL cell line (2-14 cells)&predominantly&expressed&the&receptor&of&R=spondin,&LGR4,&
whereas&we&found&little&expression&of&LGR5&and&LGR6. Alizarin red S staining and von Kossa
staining revealed that RSPO2 stimulation significantly enhanced osteoblastic differentiation of 2-14
cells&cultured&in&osteogenic&induction&medium. In addition, DKK1, an inhibitor of the canonicalWnt/β-catenin signaling pathway, suppressed the RSPO2-induced osteoblastic differentiation of
2-14 cells.
These results suggest that RSPO2 enhanced osteoblastic differentiation of immature human
PDL cells through the canonical Wnt signaling pathway. Therefore, RSPO2 may be a therapeutic
molecule in PDL healing or regeneration.
Key words: canonical Wnt/β-catenin signaling, immature human periodontal ligament cell line,
osteoblastic differentiation, R-spondin2

41

PSS-5
The roles of miR875-5p as a specific marker
during early tooth development
Keita Funada, Keigo Yosizaki, Xue Han, Kanako Miyazaki, Chieko Arai,
Tomomi Yuta, Ichiro Takahashi
Section of Orthodontics and Dentofacial Orthopedics, Kyushu University, Fukuoka 812-8582,
Japan
Epithelial–mesenchymal interaction plays critical roles for the development of organs such as
tooth, lung, salivary gland, kidney and hair. During this process, epithelial thickening and budding
into mesenchyme are common phenomenon, while the decision of cell fate is made by the specifictranscriptional controls of individual organs. We explored specific transcriptional start sites (TSS)of each organ by CAGE (Cap Analysis of Gene Expression) analysis using embryonic day 11 (E11),
E12, E13 and E14 ICR mouse. We identified a tooth specific TSS, which has been detected on
chromosome 15qD1 region. This TSS potentially codes both microRNA 599 (miR599) and miR875.
To identify which microRNA is coded from the TSS during tooth development, we performed
RT-qPCR to determine the expression level of miR599 and miR875 at early developmental stage of
tooth. According to this assay, we found that miR875-5p is specifically expressed in E14 tooth germ.
We also tested the expression pattern of miR875-5p in tooth development from E11 to postnatal day
7 (P7). We found that miR875-5p was highly expressed in E14 tooth germ and the expression
decreased gradually after that. In addition, we found that miR875-5p was expressed in mesenchymeof E14 tooth germ by RT-qPCR and in situ hybridization. To assess the role of miR875-5p in dental
mesenchyme during tooth development, we transfected mimic of miR875-5p into mouse dental
pulpal (mDP) cells, which do not express miR875-5p originally. We identified that the expressionof odontoblast marker, Dspp (dentin sialophosphoprotein) was significantly induced by miR875-5p
in mDP cells. In this study, we identified tooth specific miRNA at early developmental stage.
miR875 may regulate odontoblast differentiation and can be used as a specific marker for earlystages of dental mesenchymal cells.
Key words: Tooth, dental mesenchymal cell, CAGE analysis, miRNA

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