Faculty Profiles - EKUNI Daisuke

MISC - EKUNI Daisuke

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大学新入生における歯肉炎の性差についての検討

古田美智子, 江國大輔, 入江浩一郎, 東哲司, 加藤朋, 山田小百合, 友藤孝明, 森田学

口腔衛生学会雑誌 60 ( 4 ) 364 - 364 2010.8

運動による肥満予防がラット歯周組織に及ぼす影響

東哲司, 入江浩一郎, 遠藤康正, 粕山健太, 加藤朋, 江國大輔, 玉木直文, 友藤孝明, 森田学

口腔衛生学会雑誌 60 ( 4 ) 414 - 414 2010.8

咬合異常は顎下腺BDNFに影響を及ぼす

入江浩一郎, 江國大輔, 友藤孝明, 遠藤康正, 東哲司, 粕山健太, 森田学

日本抗加齢医学会総会プログラム・抄録集 10回 236 - 236 2010.5

Experimental Periodontitis Induces Gene Expression of Proinflammatory Cytokines in Liver and White Adipose Tissues in Obesity

Yasumasa Endo, Takaaki Tomofuji, Daisuke Ekuni, Koichiro Irie, Tetsuji Azuma, Naofumi Tamaki, Tatsuo Yamamoto, Manabu Morita

JOURNAL OF PERIODONTOLOGY 81 ( 4 ) 520 - 526 2010.4

Language：English Publisher：AMER ACAD PERIODONTOLOGY

Background: Recent studies indicated that periodontitis induces systemic low-grade inflammation. The increase in systemic low-grade inflammation induced by periodontitis may alter the effects of obesity on the production of inflammatory molecules, including C-reactive protein (CRP), interleukin (IL)-6, and tumor necrosis factor-alpha (TNF-alpha), in the liver and white adipose tissue (WAT). The purpose of the present study is to investigate the effects of periodontitis on the expression of proinflammatory cytokines in the liver and WAT in obese Zucker rats.
Methods: Obese Zucker rats and their lean litter mates were divided into four groups of six rats each: lean Zucker rats without periodontitis (control group), lean Zucker rats with periodontitis (periodontitis group), obese Zucker rats without periodontitis (obesity group), and obese Zucker rats with periodontitis (combination group). Periodontitis was ligature induced for 4 weeks in the periodontitis and combination groups, whereas the other groups were left unligated.
Results: At 4 weeks, the gene expression for CRP, IL-6, and TNF-alpha in the liver and CRP and IL-6 in the WAT of combination groups was significantly higher than in each of the three groups. Serum TNF-alpha in the periodontitis and obesity groups was significantly higher than in the control group. Serum CRP and TNF-alpha in the combination group was significantly higher than in each of the three groups.
Conclusion: Systemic low-grade inflammation after experimental periodontitis was associated with increased gene expression for hepatic levels of TNF-alpha and CRP and adipose tissue levels of IL-6 and CRP in the obese-rat model. J Periodontol 2010;81:520-526.

DOI： 10.1902/jop.2009.090574

Effects of periodontitis on aortic insulin resistance in an obese rat model

Daisuke Ekuni, Takaaki Tomofuji, Koichiro Irie, Kenta Kasuyama, Michihiro Umakoshi, Tetsuji Azuma, Naofumi Tamaki, Toshihiro Sanbe, Yasumasa Endo, Tatsuo Yamamoto, Takashi Nishida, Manabu Morita

LABORATORY INVESTIGATION 90 ( 3 ) 348 - 359 2010.3

Language：English Publisher：NATURE PUBLISHING GROUP

The combination of obesity and its associated risk factors, such as insulin resistance and inflammation, results in the development of atherosclerosis. However, the effects of periodontitis on atherosclerosis in an obese body remain unclear. The aim of the study was to investigate the effects of ligature-induced periodontitis in Zucker fatty rats on initiation of atherosclerosis by evaluating aortic insulin resistance. Zucker fatty rats (n = 24) were divided into two groups. In the periodontitis group, periodontitis was ligature-induced for 4 weeks, whereas the control group was left unligated. After the 4-week experimental period, descending aorta was used for measuring the levels of lipid deposits, immunohistochemical analysis, and evaluation of gene expression. Levels of serum C-reactive protein (CRP), tumor necrosis factor-alpha (TNF-alpha), and insulin were also measured. Rats in the periodontitis group had significantly enhanced lipid deposits in the aorta, but not in the control group. Expression of suppressor of cytokine signaling 3, vascular cell adhesion molecule 1, reactive oxygen species, nitrotyrosine, and endothelin-1 in the periodontitis group was more intense than that in the control group. Significantly decreased levels of phosphatidylinositol 3-kinase (Pi3k) catalytic beta-polypeptide (Pi3kcb), Pi3kp85, and insulin receptor substrate 1 and 2 were observed in the periodontitis group. Levels of serum CRP and TNF-alpha were significantly increased in the periodontitis group. Under insulin-stimulated conditions, aorta in the periodontitis group altered the Akt phosphorylation. Periodontitis in obesity induced the initial stage of atherosclerosis and disturbed aortic insulin signaling.

DOI： 10.1038/labinvest.2009.141

Effects of topical application of lipopolysaccharide and proteases on hepatic injury induced by high-cholesterol diet in rats

T. Yamamoto, T. Tomofuji, N. Tamaki, D. Ekuni, T. Azuma, T. Sanbe

JOURNAL OF PERIODONTAL RESEARCH 45 ( 1 ) 129 - 135 2010.2

Language：English Publisher：WILEY-BLACKWELL PUBLISHING, INC

Background and Objective: Topical application of lipopolysaccharide and proteases to the gingival sulcus induced not only periodontal inflammation but also detectable liver changes in rats fed a normal diet. However, these changes in the liver were not sufficient to induce pathological consequences. The purpose of the present study was to investigate whether gingival inflammation-induced liver change would have more dramatic pathological consequences in rats fed a high-cholesterol diet compared with the effect of the high-cholesterol diet alone.
Material and Methods: Twenty-four male Wistar rats were divided into four groups. During an 8 week experimental period, two groups were fed a normal diet and the other two were fed a high-cholesterol diet containing 1% cholesterol (w/w) and 0.5% cholic acid (w/w). Four weeks prior to the end of the experimental period, one of each of the dietary groups received daily topical application of lipopolysaccharide and proteases to the gingival sulcus, while the other was treated with pyrogen-free water.
Results: In the rats without application of lipopolysaccharide and proteases, the serum level of hexanoyl-lysine, scores of steatosis and inflammation, and concentration of 8-hydroxydeoxyguanosine in liver of rats fed a high-cholesterol diet were higher than in those fed a normal diet. In rats fed a high-cholesterol diet, the scores of steatosis and inflammation and the concentration of 8-hydroxydeoxyguanosine in the liver of rats with application of lipopolysaccharide and proteases were higher than in those without.
Conclusion: In a rat model, application of lipopolysaccharide and proteases to the gingival sulcus augmented the effect of a high-cholesterol diet on steatosis, inflammation and oxidative damage in the liver.

DOI： 10.1111/j.1600-0765.2009.01212.x

Effects of mechanical stimulation by a powered toothbrush on the healing of periodontal tissue in a rat model of periodontal disease

D. Ekuni, R. Yamanaka, T. Yamamoto, M. Miyauchi, T. Takata, T. Watanabe

JOURNAL OF PERIODONTAL RESEARCH 45 ( 1 ) 45 - 51 2010.2

Language：English Publisher：WILEY-BLACKWELL PUBLISHING, INC

Background and Objective: Elimination of pathogens is the main aim of periodontal treatment; however, modulation of the host immune response should also be considered. This study aimed to evaluate the effects of mechanical stimulation on periodontal healing in rats.
Material and Methods: Before starting the experiment, lipopolysaccharide and proteases were applied once a day, for 4 wk, to both maxillary first molars of 30 rats to induce periodontal disease, and the application was stopped at the end of the 4-wk period. The experiment started immediately following this pretreatment. In the experiment, the left palatal gingiva was stimulated once daily using a powered toothbrush and the right gingiva served as a control (no mechanical stimulation). Pathological changes, and proliferation and cell death in periodontal tissues, were evaluated histometrically and immunohistochemically at baseline (0 wk), and at 1 and 3 wk of stimulation.
Results: The control showed a reduction of polymorphonuclear leukocyte infiltration in connective tissue and an increase in the numbers of gingival and periodontal ligament fibroblasts. Mechanical stimulation reduced polymorphonuclear leukocyte infiltration and the area of destroyed collagen in connective tissue, and increased the number of gingival fibroblasts; however, it had no effect on alveolar bone and root resorption or on the number of periodontal ligament fibroblasts.
Conclusion: Mechanical stimulation accelerated the healing of gingival inflammation by reducing the infiltration of polymorphonuclear leukocytes and enhancing fibroblast proliferation and collagen synthesis.

DOI： 10.1111/j.1600-0765.2009.01195.x

Imbalance of oxidative/anti-oxidative status induced by periodontitis is involved in apoptosis of rat submandibular glands International journal

Daisuke Ekuni, Yasumasa Endo, Koichiro Irie, Tetsuji Azuma, Naofumi Tamaki, Takaaki Tomofuji, Manabu Morita

ARCHIVES OF ORAL BIOLOGY 55 ( 2 ) 170 - 176 2010.2

Language：English Publisher：PERGAMON-ELSEVIER SCIENCE LTD

Objective: Epidemiologic studies suggest a relationship between periodontitis and salivary gland dysfunction. A rat periodontitis model was used to investigate whether a causal relationship exists between periodontitis and pathological changes of submandibular glands.
Design: Fourteen male Wistar rats (8 weeks old) were divided into two groups (n = 7/group): a control group and periodontitis group. Periodontitis was induced by ligature placement around the mandibular first molars. Serum levels for reactive oxygen metabolites, antioxidant and tumour necrosis factor (TNF)-alpha were measured at baseline, 2 and 4 weeks. At 4 weeks, the levels of 8-hydroxydeoxyguanosine were determined to evaluate oxidative damage of submandibular glands. Expression of TNF-alpha mRNA and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) as well as histological findings were also evaluated in the submandibular glands.
Results: The rats with experimental periodontitis showed increase in the levels of serum reactive oxygen metabolites and TNF-alpha, and a decrease of anti-oxidant power in a time-dependent manner. At 4 weeks, these rats also had significantly increased levels of 8-hydroxydeoxyguanosine and TNF-alpha, and increased number of TUNEL-positive cells and vacuolisation in the submandibular glands compared to the control rats.
Conclusions: Imbalance of circulating oxidative/anti-oxidative status may be involved in vacuolisation and apoptosis of submandibular glands in the rat periodontitis model. (C) 2009 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.archoralbio.2009.11.013

Relationship between periodontal status and components of metabolic syndrome in a rural Japanese population

YAMAMOTO T

J Dent Hlth 60 96 - 103 2010

ワークショップ「頭頸部がんチーム医療における口腔ケアの促進」と連携強化．

山中玲子, 丸山貴之, 横井彩, 山田小百合, 玉木直文, 江國大輔, 友藤孝明, 森田学

岡山歯学会雑誌 61 2010

メインテナンス期の歯周病患者における咬合力と歯周病進行との関係について

竹内倫子, 江國大輔, 山本龍生, 森田学

岡山歯学会雑誌 29 ( 1 ) 67 - 67 2010

健康診査データにおけるメタボリックシンドロームと歯周炎の関係

山本龍生, 恒石美登里, 古田美智子, 小山玲子, 江國大輔, 森田学, 平田幸夫

神奈川歯学 44 ( 抄録集 ) 25 - 25 2009.12

Preventive Effects of a Cocoa-Enriched Diet on Gingival Oxidative Stress in Experimental Periodontitis

Takaaki Tomofuji, Daisuke Ekuni, Koichiro Irie, Tetsuji Azuma, Yasumasa Endo, Naofumi Tamaki, Toshihiro Sanbe, Jun Murakami, Tatsuo Yamamoto, Manabu Morita

JOURNAL OF PERIODONTOLOGY 80 ( 11 ) 1799 - 1808 2009.11

Language：English Publisher：AMER ACAD PERIODONTOLOGY

Background: Oxidative stress affects the progression of periodontitis. Cocoa is a rich source of flavonoids with antioxidant properties, which could suppress gingival oxidative stress in periodontal lesions. The purpose of the present study was to investigate the effects of a cocoa-enriched diet on gingival oxidative stress in a rat-periodontitis model.
Methods: In this 4-week study, rats were divided into three groups (n = 8/group): a control group (fed a regular diet) and two periodontitis groups (fed a regular diet or cocoa-enriched diet [10% of food intake]). Periodontitis was induced by ligature placement around the mandibular first molars. Serum levels for reactive oxygen metabolites were measured at baseline and 2 and 4 weeks. At 4 weeks, the levels of 8-hydroxydeoxyguanosine and reduced/oxidized glutathione ratio were determined to evaluate gingival oxidative damage and antioxidant status, respectively.
Results: Rats with experimental periodontitis that were fed a regular diet showed an increase in the level of serum reactive oxygen metabolites in a time-dependent manner. These rats also had an increased 8-hydroxydeoxyguanosine level and decreased reduced/oxidized glutathione ratio in the gingival tissue, inducing alveolar bone loss and polymorphonuclear leukocyte infiltration. Although experimental periodontitis was induced in the rats fed a cocoa-enriched diet, they did not show impairments in serum reactive oxygen metabolite level and gingival levels for 8-hydroxydeoxyguanosine and reduced/oxidized glutathione ratio. Alveolar bone loss and polymorphonuclear leukocyte infiltration after ligature placement were also inhibited by cocoa intake.
Conclusion: Consuming a cocoa-enriched diet could diminish periodontitis-induced oxidative stress, which, in turn, might suppress the progression of periodontitis. J Periodontol 2009;80:1799-1808.

DOI： 10.1902/jop.2009.090270

急性期病院における頭頸部癌患者への口腔ケア

山中玲子, 小山玲子, 江國大輔, 古田美智子, 丸山貴之, 山本龍生, 森田学

口腔衛生学会雑誌 59 ( 5 ) 609 - 610 2009.10

Lipopolysaccharide-Induced Epithelial Monoamine Oxidase Mediates Alveolar Bone Loss in a Rat Chronic Wound Model International journal

Daisuke Ekuni, James D. Firth, Tarun Nayer, Takaaki Tomofuji, Toshihiro Sanbe, Koichiro Irie, Tatsuo Yamamoto, Takashi Oka, Zhenzi Liu, Juergen Vielkind, Edward E. Putnins

AMERICAN JOURNAL OF PATHOLOGY 175 ( 4 ) 1398 - 1409 2009.10

Language：English Publisher：AMER SOC INVESTIGATIVE PATHOLOGY, INC

Reactive oxygen species (ROS) production is an antimicrobial response to pathogenic challenge that may, in the case of persistent infection, have deleterious effects on the tissue of origin. A rat periodontal disease model was used to study ROS-induced chronic epithelial inflammation and bone loss. Lipopolysaccharide (LPS) was applied for 8 weeks into the gingival sulcus, and histological analysis confirmed the onset of chronic disease. junctional epithelium was collected from healthy and diseased animals using laser-capture microdissection, and expression microarray analysis was performed. Of 19,730 genes changed in disease, 42 were up-regulated >= 4-fold. Three of the top 10 LPS-induced genes, monoamine oxidase B (MAO/B) and flavin-containing monooxygenase 1 and 2, are implicated in ROS signaling. LPS-associated induction of the ROS mediator H(2)O(2) as well as MAO/B and tumor necrosis factor (TNF)-alpha levels were validated in the rat histological sections and a porcine junctional epithelial cell culture model. Topical MAO inhibitors significantly counteracted LPS-associated elevation of H(2)O(2) production and TNF-alpha expression in vivo and in vitro, inhibited disease-associated apical migration and proliferation of junctional epithelium and inhibited induced systemic H(2)O(2) levels and alveolar bone loss in vivo. These results suggest that LPS induces chronic wounds via elevated MAO/B-mediated increases in H(2)O(2) and TNF-alpha activity by epithelial cells and is further associated with more distant effects on systemic oxidative stress and alveolar bone loss. (Am J Pathol 2009,175:1398-1409; DOI: 10.2353/ajpath.2009.090108)

DOI： 10.2353/ajpath.2009.090108

A single application of hydrogen sulphide induces a transient osteoclast differentiation with RANKL expression in the rat model International journal

Koichiro Irie, Daisuke Ekuni, Tatsuo Yamamoto, Manabu Morita, Ken Yaegaki, Hisataka Ii, Toshio Imai

ARCHIVES OF ORAL BIOLOGY 54 ( 8 ) 723 - 729 2009.8

Language：English Publisher：PERGAMON-ELSEVIER SCIENCE LTD

Objective: Oral malodor is mainly attributed to volatile sulphur compounds (VSCs) such as hydrogen sulphide (H2S), methyl mercaptan and dimethyl sulphide. VSC accelerate periodontal soft tissue destruction. However, there is little information about the potential role of H2S in alveolar bone loss. The purpose of this animal study was to examine the effects of sodium hydrogen sulphide (NaHS), H2S donor drug, on osteoclast differentiation in rat periodontal tissue.
Design: Twenty-four male Wistar rats (8 weeks old) were divided into four groups: a control group and three experimental groups, which were examined at 3 h, 1 day, and 3 days after topical application of 3 mu l NaHS (l M in physiological saline) into the gingival sulcus of rat first molar. Expression of tumour necrosis factor (TNF)-alpha, RANKL, NF-kappa B and tartrate-resistant acid phosphatase (TRAP) was evaluated in the periodontal tissue.
Results: Three hours after NaHS application, TNF-alpha expression increased in the periodontal ligament. The numbers of RANKL-positive osteoblasts and TRAP-positive osteoclasts significantly increased progressively with time and reached a maximum level after 1 day. Significant up-regulation of RANKL and NF-kappa B mRNA was observed at 3 h after NaHS application.
Conclusions: H2S application caused a transient increase of osteoclast differentiation with up-regulation of RANKL expression in osteoblasts. H2S, which is primarily responsible for halitosis, may also contribute to alveolar bone resorption through RANKL expression. (C) 2009 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.archoralbio.2009.05.006

Periodontitis-induced lipid peroxidation in rat descending aorta is involved in the initiation of atherosclerosis

D. Ekuni, T. Tomofuji, T. Sanbe, K. Irie, T. Azuma, T. Maruyama, N. Tamaki, J. Murakami, S. Kokeguchi, T. Yamamoto

JOURNAL OF PERIODONTAL RESEARCH 44 ( 4 ) 434 - 442 2009.8

Language：English Publisher：WILEY-BLACKWELL PUBLISHING, INC

Background and Objective:
Periodontitis is a risk factor for the development of atherosclerosis. Recent studies indicate that oxidative mechanisms, including lipid peroxidation, are involved not only in periodontitis but also in atherosclerosis. Lipid peroxidation may play an important role in the pathogenesis of atherosclerosis, particularly during its earliest stages. The purpose of this study was to investigate the relationship between lipid peroxidation induced by periodontitis and the initiation of atherosclerosis.
Material and Methods:
Sixteen rats were randomly divided into two groups of eight rats each. Periodontitis was ligature-induced for 4 wk in the experimental group, whereas the control group was left untreated. After the experimental period, the mandibular first molar regions were resected and then subjected to histological analysis and measurement of hexanoyl-lysine expression as an indicator of lipid peroxidation. Descending aorta was used for measuring the levels of hexanoyl-lysine, reactive oxygen species and lipid deposits, and for real-time polymerase chain reaction microarray analysis. The level of hexanoyl-lysine was also measured in serum.
Results:
In the experimental group, the levels of hexanoyl-lysine in periodontal tissue and serum increased. Only aorta samples in the experimental group showed lipid accumulation, with increased expression of hexanoyl-lysine, reactive oxygen species and oxidative stress-related genes (including nitric oxide synthases 2 and 3), whereas the superoxide dismutase 1 gene level was down-regulated.
Conclusion:
In a ligature-induced periodontitis rat model, increased lipid peroxidation was found in serum and aorta as well as in periodontal tissue. Atherosclerosis-related gene expression and histological changes were also stimulated. Periodontitis-induced lipid peroxidation in the aorta may be involved in the early stage of atherosclerosis.

DOI： 10.1111/j.1600-0765.2008.01122.x

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Relation of dietary preference to bite force and occlusal contact area in Japanese children

R. Yamanaka, R. Akther, M. Furuta, R. Koyama, T. Tomofuji, D. Ekuni, N. Tamaki, T. Azuma, T. Yamamoto, E. Kishimoto

JOURNAL OF ORAL REHABILITATION 36 ( 8 ) 584 - 591 2009.8

Language：English Publisher：WILEY-BLACKWELL PUBLISHING, INC

P>The purpose of this study was to examine the relationship of dietary preference to bite force and occlusal contact area in Japanese elementary school children. A total of 348 children, aged 7-12 years, from two public elementary schools located in Okayama Prefecture, Japan, participated in the study. Clinical examination included decayed, missing and filled teeth (dmft and DMFT), and total numbers of deciduous and permanent teeth. Bite force and occlusal contact area were measured using a pressure-detecting sheet. Dietary preference was assessed using a questionnaire in which the answers were given in like/dislike form. Mann-Whitney U-test and multiple logistic regression analysis were applied to analyse the data. In multiple logistic regression analysis after adjustment for age, gender and total number of teeth present, children who liked cabbage and celery showed significantly higher bite force (P = 0 center dot 05 and P < 0 center dot 01, respectively) than those who disliked these. Children who liked cabbage and celery also showed higher occlusal contact area (P < 0 center dot 05 and P < 0 center dot 01, respectively) than those who disliked these. The Japanese elementary school children who liked hard foods such as cabbage and celery showed higher bite force and higher occlusal contact area than those who disliked these foods. A positive attitude towards harder food items might contribute to healthy development of the masticatory apparatus.

DOI： 10.1111/j.1365-2842.2009.01971.x

Effects of Obesity on Gingival Oxidative Stress in a Rat Model International journal

Takaaki Tomofuji, Tatsuo Yamamoto, Naofumi Tamaki, Daisuke Ekuni, Tetsuji Azuma, Toshihiro Sanbe, Koichiro Irie, Kenta Kasuyama, Michihiro Umakoshi, Jun Murakami, Susumu Kokeguchi, Manabu Morita

JOURNAL OF PERIODONTOLOGY 80 ( 8 ) 1324 - 1329 2009.8

Language：English Publisher：AMER ACAD PERIODONTOLOGY

Background: Studies indicate a correlation between obesity and periodontitis. Oxidative stress is involved in the progression of periodontitis. The purpose of this study was to investigate the effects of obesity on gingival oxidative stress in a rat periodontitis model.
Methods: The obese Zucker rats (n = 14) and their lean littermates (n = 14) were each divided into two groups of seven rats. In one of each group, periodontitis was induced by ligature for 4 weeks, whereas the other group was left unligated. The level of 8-hydroxydeoxyguanosine and the ratio of reduced/oxidized glutathione were determined to examine gingival oxidative stress. The serum level of reactive oxygen metabolites and the gingival gene-expression pattern related to oxidative/metabolic stress, inflammation, and cell behavior were also evaluated.
Results: The obese rats weighed more than the lean rats at 4 weeks. Compared to lean rats, obese rats had enhanced gingival 8-hydroxydeoxyguanosine levels and a decreased ratio of reduced/oxidized glutathione in the gingival tissue, with increasing serum reactive oxygen metabolites. However, there were no significant differences in the degree of alveolar bone loss between lean and obese rats, except for teeth with and without ligatures in both rats. In addition, the periodontal lesion in obese rats showed higher 8-hydroxydeoxyguanosine levels and polymorphonuclear leukocyte infiltration than the inflamed ones in lean rats, with downregulation of multiple cytochrome P450 gene expression.
Conclusions: Obesity induced gingival oxidative stress with increasing serum reactive oxygen metabolites in rats. In the periodontal lesion, gene expressions related to a capacity for xenobiotic detoxification were downregulated in the obese model. J Periodontol 2009;80:1324-1329.

DOI： 10.1902/jop.2009.080621

Periodontitis-induced lipid peroxidation in rat descending aorta is involved in the initiation of atherosclerosis

D. Ekuni, T. Tomofuji, T. Sanbe, K. Irie, T. Azuma, T. Maruyama, N. Tamaki, J. Murakami, S. Kokeguchi, T. Yamamoto

JOURNAL OF PERIODONTAL RESEARCH 44 ( 4 ) 434 - 442 2009.8

Language：English Publisher：WILEY-BLACKWELL PUBLISHING, INC

DOI： 10.1111/j.1600-0765.2008.01122.x

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