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Intragingival injection of Porphyromonas gingivalis-derived lipopolysaccharide induces a transient increase in gingival tumour necrosis factor-a, but not interleukin-6,in anaesthetised rats

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International Journal of Oral Science 2015年第3期7卷 155-160页

作者： Hiroko Taguchi Yuri Aono Takayuki Kawato Masatake Asano Noriyoshi Shimizu Tadashi Saigusa department of Orthodontics Nihon University School of Dentistry department of Pharmacology Nihon University School of Dentistry at Matsudo department of Pharmacology Nihon University School of Dentistry department of Oral Health Sciences Nihon University School of Dentistry department of Pathology Nihon University School of Dentistry

This study used in vivo microdialysis to examine the effects of intragingival application of lipopolysaccharide（LPS） derived from Porphyromonas gingivalis（Pg-LPS） on gingival tumour necrosis factor（TNF）-a and interleukin（IL）-6 levels in rats. A microdialysis probe with an injection needle attached to the surface of the dialysis membrane was implanted into the gingiva of the upper incisor. For comparison, the effects of LPS derived from Escherichia coli（Ec-LPS） on IL-6 and TNF-a levels were also analysed. Pg-LPS（1 mg/1 m L） or Ec-LPS（1 or 6 mg/1 m L） was applied by microsyringe, with gingival dialysates collected every hour. Enzyme-linked immunosorbent assay（ELISA） revealed that gingival dialysates contained approximately 389 pg？m L21 of IL-6 basally; basal TNF-a levels were lower than the detection limit of the ELISA. Pg-LPS failed to alter IL-6 levels but markedly increased TNF-a levels, which remained elevated for 2 h after treatment. Neither IL-6 nor TNF-a were affected by Ec-LPS. Reverse transcriptase-polymerase chain reaction（RT-PCR） analysis revealed that the gingiva expresses Toll-like receptor（TLR） 2 and TLR4 m RNA. Immunohistochemical examination showed that TLR2 and TLR4 are expressed by gingival epithelial cells. The present study provides in vivo evidence that locally applied Pg-LPS, but not Ec-LPS, into the gingiva transiently increases gingival TNF-a without affecting IL-6. The present results suggest that TLR2 but not TLR4 expressed on gingival epithelial cells may mediate the Pg-LPS-induced increase in gingival TNF-a in rats.

关键词： Porphyromonas gingivalis lipopolysaccharide gingiva tumour necrosis factor-a microdialysis

Enamel Matrix Derivative Stimulates Osteogenesis-and Chondrogenesis-related Transcription Factors in C3H10T1/2 Cells

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Acta Biochimica et Biophysica Sinica 2007年第1期39卷 1-7页

作者： Masataka NARUKAWA Naoto SUZUKI Tadahiro TAKAYAMA Yasuhiko YAMASHITA Kichibee OTSUKA Koichi ITO department of periodontology Nihon University School of DentistryTokyo101-8310Japan department of Biochemistry Nihon University School of DentustryTokyo 101-8310Japan Division of Functional Morphology Dental Research CenterNihon University School of DentistryTokyo 101-8310Japan department of Biochemistry Nihon University School of DentistryTokyo101-8310Japan Division of Functional Morphology Dental Research CenterNihon University School of DentistryTokyo101-8310Japan Division of Advanced Dental Treatment Dental Research CenterNihon University School of DentistryTokyo101-8310Japan

Our purpose was to determine how enamel matrix derivative (EMD) affects the expression ofosteogenesis- and chondrogenesis-related transcription factors in undifferentiated mesenchymal cells.C3H10T1/2 cell line,a typical pluripotential mesenchymal cell line,was cultured with or without EMD forup to 7 *** of mRNAs encoding osteogenesis-and chondrogenesis-related transcription factors(Runx2, Osterix,AJ18,Dlx5,Msx2,Sox5,Sox9 and Zfp60) was measured using real-time polymerasechain ***2 and Sox9 protein expression and the presence of bone morphogenetic protein (BMP)-6-like molecules in EMD were determined by Western *** substantially increased mRNA levels ofosteogenesis-and chondrogenesis-related transcription *** also induced Runx2 and Sox9 *** blotting analysis of EMD using anti-BMP-6 antibody revealed immunoreactive bandscorresponding to about 14 kDa and 60 *** results suggest that EMD stimulates osteogenesis- andchondrogenesis-related transcription factors,and these activities may be mediated,at least in part,by BMP-6in EMD.

关键词： enamel matrix derivative osteogenesis chondrogenesis

Micromolar sodium fluoride mediates anti-osteoclastogenesis in Porphyromonas gingivalis-induced alveolar bone loss

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International Journal of Oral Science 2015年第4期7卷 242-249页

作者： Ujjal K Bhawal Hye-Jin Lee Kazumune Arikawa Michiharu Shimosaka Masatoshi Suzuki Toshizo Toyama Takenori Sato Ryota Kawamata Chieko Taguchi Nobushiro Hamada Ikuo Nasu Hirohisa Arakawa Koh Shibutani department of Biochemistry and Molecular Biology Nihon University School of Dentistry at Matsudo Research Institute of Oral Health Nihon University School of Dentistry at Matsudo department of Oral Health Graduate School of DentistryKanagawa Dental University department of Dental Hygiene Howon University department of Preventive and Public Oral Health Nihon University School of Dentistry at Matsudo department of Anesthesiology Nihon University School of Dentistry at Matsudo department of Infection Control Division of MicrobiologyKanagawa Dental University department of Radiopraxis Science Kanagawa Dental University

Osteoclasts are bone-specific multinucleated cells generated by the differentiation of monocyte/macrophage lineage precursors. Regulation of osteoclast differentiation is considered an effective therapeutic approach to the treatment of bone-lytic diseases. Periodontitis is an inflammatory disease characterized by extensive bone resorption. In this study, we investigated the effects of sodium fluoride （NaF） on osteoclastogenesis induced by Porphyromonas gingivalis, an important colonizer of the oral cavity that has been implicated in periodontitis. NaF strongly inhibited the P. gingivalis-induced alveolar bone loss. That effect was accompanied by decreased levels of cathepsin K, interleukin （IL）-1β, matrix metalloproteinase 9 （MMP9）, and tartrate-resistant acid phosphatase, which were up-regulated during P. gingivalis-induced osteoclastogenesis. Consistent with the in vivo anti-osteoclastogenic effect, NaF inhibited osteoclast formation caused by the differentiation factor RANKL （receptor activator of nuclear factor KB ligand） and macrophage colony-stimulating factor （M-CSF）. The RANKL-stimulated induction of the transcription factor nuclear factor of activated T cells （NFAT） cl was also abrogated by NaF. Taken together, our data demonstrate that NaF inhibits RANKL-induced osteoclastogenesis by reducing the induction of NFATcl, ultimately leading to the suppressed expression of cathepsin K and MMP9. The in vivo effect of NaF on the inhibition of P. gingivalis-induced osteoclastogenesis strengthens the potential usefulness of NaF for treating periodontal diseases.

关键词： alveolar bone loss osteoclasts Porphyromonas gingivalis sodium fluoride

Juxtacrine interaction of macrophages and bone marrow stromal cells induce interleukin-6 signals and promote cell migration

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Bone Research 2015年第2期3卷 100-111页

作者： Jia Chang Amy J Koh Hernan Roca Laurie K McCauley department of Periodontics and Oral Medicine University of Michigan School of Dentistry department of periodontology University of Florida College of Dentistry department of Pathology University of Michigan Medical School

The bone marrow contains a heterogeneous milieu of cells, including macrophages, which are key cellular mediators for resolving infection and inflammation. Macrophages are most well known for their ability to phagocytose foreign bodies or apoptotic cells to maintain homeostasis; however, little is known about their function in the bone microenvironment. In the current study, we investigated the in vitro interaction of murine macrophages and bone marrow stromal cells （BMSCs）, with focus on the juxtacrine induction of IL-6 signaling and the resultant effect on BMSC migration and growth. The juxtacrine interaction of primary mouse macrophages and BMSCs activated IL-6 signaling in the co-cultures, which subsequently enhanced BMSC migration and increased BMSC numbers. BMSCs and macrophages harvested from IL-6 knockout mice revealed that IL-6 signaling was essential for enhancement of BMSC migration and increased BMSC numbers via juxtacrine interactions. BMSCs were the main contributor of IL-6 signaling~ and hence activation of the IL-6/ gp130/STAT3 pathway. Meanwhile, macrophage derived IL-6 remained important for the overall production of IL-6 protein in the co-cultures. Taken together, these findings show the function of macmphages as co-inducers of migration and growth of BMSCs, which could directly influence bone formation and turnover.

关键词： BMSCs IL bone STAT cell

Caffeic acid phenethyl ester protects against oxidative stress and dampens inflammation via heme oxygenase 1

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International Journal of Oral Science 2019年第1期11卷 50-57页

作者： Alexandra St?hli Ceeneena Ubaidha Maheen Franz Josef Strauss Sigrun Eick Anton Sculean Reinhard Gruber department of Oral Biology School of Dentistry Medical University of Vienna department of periodontology School of Dental MedicineUniversity of Bern department of Conservative dentistry School of Dentistry University of Chile Austrian Cluster for Tissue Regeneration

Periodontal disease is associated with chronic oxidative stress and inflammation. Caffeic acid phenethyl ester(CAPE), which is a potent inducer of heme oxygenase 1(HO1), is a central active component of propolis, and the application of propolis improves periodontal status in diabetic patients. Here, primary murine macrophages were exposed to CAPE. Target gene expression was assessed by whole-genome microarray, RT-PCR and Western blotting. The antioxidative and anti-inflammatory activities of CAPE were examined by exposure of the cells to hydrogen peroxide, saliva and periodontal pathogens. The involvement of HO1 was investigated with the HO1 inhibitor tin protoporphyrin(SnPP) and knockout mice for Nrf2, which is a transcription factor for detoxifying enzymes. CAPE increased HO1 and other heat shock proteins in murine macrophages. A p38 MAPK inhibitor and Nrf2 knockout attenuated CAPE-induced HO1 expression in macrophages. CAPE exerted strong antioxidative activity. Additionally, CAPE reduced the inflammatory response to saliva and periodontal pathogens. Blocking HO1 decreased the antioxidative activity and attenuated the anti-inflammatory activity of CAPE. In conclusion, CAPE exerted its antioxidative effects through the Nrf2-mediated HO1 pathway and its anti-inflammatory effects through NF-κB inhibition. However, preclinical models evaluating the use of CAPE in periodontal inflammation are necessary in future studies.

关键词： Caffeic acid phenethyl ester(CAPE) inflammatory response

Analysis of dynamic smile and upper lip curvature in young Chinese

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International Journal of Oral Science 2013年第1期5卷 49-53页

作者： Ling-Zhi Liang Wen-Jie Hu Yan-Ling Zhang Kwok-Hung Chung department of periodontology Peking University School & Hospital of Stomatology department of Restorative dentistry University of Washington

During smile evaluation and anterior esthetic construction,the anatomic and racial variations should be considered in order to achieve better matching *** aims of this study were to validate an objective method for recording spontaneous smile process and to categorize the smile and upper lip curvature of Chinese Han-nationality *** hundred and eighty-eight Chinese Han-nationality youths(88 males and 100 females) ranged from 20 to 35 years of age were *** smiles were elicited by watching comical movies and the dynamics of the spontaneous smile were captured continuously with a digital video *** subjects’ smiles were categorized into three types:commissure,cuspid and gummy smile based on video editing software and final ***’ upper lip curvatures were also measured and divided into three groups:upward,straight and *** analysis was conducted to obtain intra-rater reliabilities on twice *** Pearson Chi-square test was used to compare differences for each parameters(α=0.05).In smile classification,60.6%commissure smile,33.5%cuspid smile and 5.9%gummy smile were *** upper lip measurement,26.1%upward,39.9%straight and 34.0%downward upper lip curvature were *** commissure smile group showed statistically significant higher percentage of straight(46.5%) and upward(40.4%) in upper lip curvatures(P<0.05),while cuspid smile group(65.1%) and gummy smile group(72.7%) showed statistically significant higher frequency in downward upper lip curvature(P<0.05).It is evident that differences in upper lip curvature and smile classification exist based on race,when comparing Chinese subjects with those of Caucasian descent,and gender.

关键词： Chinese Han nationality image processing smiles upper lip curvature

Lipopolysaccharide Enhances the Production of Nicotine-Induced Prostaglandin E2 by an Increase in Cyclooxygenase-2 Expression in Osteoblasts

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Acta Biochimica et Biophysica Sinica 2007年第3期39卷 163-172页

作者： Maiko SHOJI Natsuko TANABE Narihiro MITSUI Naoto SUZUKI Osamu TAKEICHI Tomoko KATONO Akira MOROZUMI Masao MAENO department of Oral Health Sciences Nihon University School of DentistryTokyo 101-8310Japan Division of Functional Morphology Dental Research Center.Nihon University School ofDentistryTokyo 101-8310Japan nihon university Grndudte school of dentistry Tokyo 101-8310Japan Division of Functional Morphology Dental Research Center.Nihon University School of DentistryTokyo 101-8310Japan department of Endodontics Nihon University School of DentisityTokyo 101-8310Japan Division of Advanced Dental Treatment Dental Research CenterNihon University School of DentistryTokyo 101-8310Japan nihon university Graduate school of dentistry Tokyo 101-8310Japan

Previous studies have indicated that lipopolysaccharide（LPS）from Gram-negative bacteria inplaque induces the release of prostaglandin E2（PGE2）,which promotes alveolar bone resorption in periodontitis,and that tobacco smoking might be an important risk factor for the development and severity of *** determined the effect of nicotine and LPS on alkaline phosphatase（ALPase）activity,PGE2production,and the expression of cyclooxygenase（COX-1,COX-2）,PGE2receptors Ep1-4,and macrophage colonystimulating factor（M-CSF）in human osteoblastic Saos-2 *** cells were cultured with 10-3M nicotinein the presence of 0,1,or 10μg/ml LPS,or with LPS *** activity decreased in cells cultured withnicotine or LPS alone,and decreased further in those cultured with both nicotine and LPS,whereas PGE2production significantly increased in the former and increased further in the *** itself,nicotine did notaffect expression of COX-1,COX-2,any of the PGE2receptors,or M-CSF,but when both nicotine and LPSwere present,expression of COX-2,Ep3,Ep4,and M-CSF increased *** addition of10-4M indomethacin eliminated the effects of nicotine and LPS on ALPase activity,PGE2production,and M-CSF *** of protein kinase A was high in cells cultured with nicotine and *** suggest that LPS enhances the production of nicotine-induced PGE2by an increase in COX-2 expres-sion in osteoblasts,that nicotine-LPS-induced PGE2interacts with the osteoblast Ep4 receptor primarily inautocrine or paracrine mode,and that the nicotine-LPS-induced PGE2then decreases ALPase activity andincreases M-CSF expression.

关键词： nicotine lipopolysaccharide prostaglandin E2 cyclooxygenase-2 Ep4 receptor

Nicotine Treatment Induces Expression of Matrix Metalloproteinases in Human Osteoblastic Saos-2 Cells

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Acta Biochimica et Biophysica Sinica 2006年第12期38卷 874-882页

作者： Tomoko KATONO Takayuki KAWATO Natsuko TANABE Naoto SUZUKI Kazuhiro YAMANAKA Hitoshi OKA Masafumi MOTOHASHI Masao MAENO Kanda Surugadai Chiyoda-kuTokyo 101-8310Japan department of Oral Health Sciences Nihon UniversitySchool of DentistryTokyo 101-8310Japan Japan department of Biochemistry Nihon University School of Dentistry.Tokyo 101-8310Japan

Tobacco smoking is an important risk factor for the development of severe ***,we showed that nicotine affected mineralized nodule formation,and that nicotine andlipopolysaccharide stimulated the formation of osteoclast-like cells by increasing production of macrophagecolony-stimulating factor （M-CSF） and prostaglandin E2（PGE2） by human osteoblastic Saos-2 *** thepresent study,we examined the effects of nicotine on the expression of matrix metalloproteinases （MMPs）,tissue inhibitors of matrix metalloproteinases （TIMPs）,the plasminogen activation system including thecomponent of tissue-type plasminogen activator （tPA）,urokinase-type PA （uPA）,and PA inhibitor type 1（PAI-1）,α7 nicotine receptor,and *** also examined the effect of the nicotine antagonist D-tubocurarineon nicotine-induced expression of *** expression was examined using real-time polymerase chainreaction （PCR） to estimate mRNA *** addition,expression of the MMP,TIMP,uPA,tPA,and PAI-1proteins was determined by Western blotting *** treatment caused expression of MMP-1,2,3,and 13,but not MMP-14,to increase significantly after 5 or 10 d of culture;MMP-14 expression did notchange through day *** of MMP-1 expression by nicotine treatment was eliminated bysimultaneous treatment with *** the presence of nicotine,expression of uPA,PAI-1,orTIMP-1,2,3,or 4 did not change over 14 d of culture,whereas expression of tPA increased significantly byday *** also increased expression of the α7 nicotine receptor and c-fos *** results suggestthat nicotine stimulates bone matrix turnover by increasing production of tPA and MMP-1,2,3,and 13,thereby tipping the balance between bone matrix formation and resorption toward the latter process.

关键词： nicotine matrix metalloproteinase tissue inhibitor of matrix metalloproteinase plasminogen activation system

Long-term follow-ups of revascularized immature necrotic teeth:three case reports

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International Journal of Oral Science 2012年第2期4卷 109-113页

作者： Duck-Su Kim Hae-Jin Park Je-Ha Yeom Ji-Sung Seo Gil-Joo Ryu Ki-Ho Park Seung-Il Shin Sun-Young Kim department of Conservative dentistry School of DentistryKyung Hee UniversitySeoulKorea Good Will Dental Hospital BusanKorea department of Orthodontics School of DentistryKyung Hee UniversitySeoulKorea department of periodontology School of DentistryKyung Hee UniversitySeoulKorea

Revascularization of immature necrotic teeth is a reliable treatment alternative to conventional apexogenesis or apexification. In case 1, a 12-year-old boy had his necrotic, immature mandibular left second premolar treated with a revascularization technique. At a 24-month follow-up, periapical radiolucency had disappeared and thickening of the root wall was observed. In cases 2 and 3, a lO-year-old boy had his necrotic, immature, bilateral mandibular second premolars treated with the same modality. At 48-month （in case 2） and 42-month （in case 3） follow-ups, loss of periapical radiolucencies and increases in the root wall thickness were also observed.

关键词： revascularization pulp necrosis mineral trioxide aggregate antibiotic mixture immature tooth