• Journal of dental hygiene science
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• v.19 no.4
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• pp.213-219
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• 2019

Background: Smoking exerts an adverse effect on the periodontal tissue by reorganizing the ecosystem of oral microorganisms and is considered to be an important factor in the development of periodontal disease. Although cross-sectional studies on smokers and non-smokers have been attempted to investigate the microbial differences in periodontal oral cavity, only few studies have been conducted to investigate the changes in oral microorganisms during smoking cessation. The purpose of this study was to investigate the changes of bacteria in saliva and gingival crevicular fluid (GCF) over a period of one year among 11 smokers trying to quit smoking. Methods: Eleven smokers trying to quit smoking visited the clinic at baseline, two weeks, two months, four months, six months, and 12 months to give saliva and GCF samples. The amounts of 16S rRNA, Porphyromonas gingivalis, Treponema denticola, Prevotella intermedia, Fusobacterium nucleatum subsp. nucleatum, Streptococcus mutans, and Streptococcus sobrinus in saliva and GCF were quantified using real-time polymerase chain reaction TaqMan probe assay. The results were analyzed by nonparametric statistical analysis using Friedman test and Spearman correlation coefficient. Results: After cessation of smoking, the amounts of 16S rRNA corresponding to P. gingivalis, F. nucleatum, P. intermedia, and T. denticola in saliva decreased and then again increased significantly. The amount of F. nucleatum 16S rRNA in GCF decreased significantly after smoking cessation. Positive correlations were observed between 16S rRNA and F. nucleatum and between F. nucleatum and T. denticola in saliva and GCF. Conclusion: Even if the number of subjects in this study was small, we suggest that smoking cessation may reduce the total bacterial amount and F. nucleatum in GCF. However, the results regarding changes in the microbial ecosystem due to smoking or smoking cessation were inconsistent. Therefore, further in-depth studies need to be carried out.

• Journal of Periodontal and Implant Science
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• v.48 no.4
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• pp.261-271
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• 2018

Purpose: Few studies have examined periodontal pathogens from saliva samples in periodontally healthy young adults. The purposes of this study were to determine the prevalence of periodontopathic bacteria and to quantify periodontal pathogens in saliva samples using real-time polymerase chain reaction (PCR) assays in periodontally healthy Korean young adults under 35 years of age. Methods: Nine major periodontal pathogens were analyzed by real-time PCR in saliva from 94 periodontally healthy young adults. Quantification of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Prevotella intermedia, Fusobacterium nucleatum, Campylobacter rectus, Peptostreptococcus anaerobius, and Eikenella corrodens was performed by DNA copy number measurement. Results: F. nucleatum and E. corrodens were detected in all subjects; the numbers of positive samples were 87 (92.6%), 91 (96.8%), and 90 (95.7%) for P. gingivalis, P. anaerobius, and C. rectus, respectively. Other pathogens were also detected in periodontally healthy subjects. Analysis of DNA copy numbers revealed that the most abundant periodontal pathogen was F. nucleatum, which was significantly more prevalent than all other bacteria (P<0.001), followed by P. anaerobius, P. gingivalis, E. corrodens, C. rectus, and T. denticola. There was no significant difference in the prevalence of each bacterium between men and women. The DNA copy number of total bacteria was significantly higher in men than in women. Conclusions: Major periodontal pathogens were prevalent in the saliva of periodontally healthy Korean young adults. Therefore, we suggest that the development of periodontal disease should not be overlooked in periodontally healthy young people, as it can arise due to periodontal pathogen imbalance and host susceptibility.

• Restorative Dentistry and Endodontics
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• v.29 no.1
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• pp.51-57
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• 2004

본 연구는 에폭시레진계 봉함제 (AH26)과 두 가지의 상아질 접착제와 함께 근관충전을 시 행하였을 때와 에폭시레진계 봉함제 단독으로 사용하였을 때의 미세누출을 혐기성 세균모델을 이용하여 평가하였다. 52개의 단근치를 이용하여 .04, .06 taper Profile (Dentsply-Maillefer, Ballaigues, Swiss)을 사용하여 변형된 crown-down pressureless법으로 근관형성 하였다. 형성된 치근을 12개씩 무작위로 나누어 4개의 실험군으로 하였으며, 나머지 치아 중 2개를 양성대조군에, 2개는 음성대조군으로 사용하였다. 제1군은 All Bond 2(Bisco, Itasca, IL, USA)를 적용하고 거타퍼쳐와 AH26 (Dentsply, Konstanz, Germany)을 이용하여 continuous wave of condensation technique으로 근관충전 하였으며, 제 2군은Prime ＆ Bond NT (Dentsply, Konstanz, Germany)를 적용 후 거타퍼쳐와 AH26을 이용하여 충전하였으며, 제3군은 17％ EDTA를 적용하여 도말층을 제거한 후 거터퍼쳐 와 AH26을 사용하여 충전하였다. 제4군은 17％ EDTA를 적용하지 않고 거터퍼쳐와 AH26을 사용하여 충전하였다. Fusobacterium nucleatum (VPI 10197)을 추적자로 이용한 혐기성세균모델을 사용하여 혐기성배양기에서 배양시키면서 60일 동안 각군에 대한 미세누출 여부를 관찰하였다. 매일 배양액의 혼탁도와 색상변화를 관찰하여 기록하였다. 제4군에서 통계학적으로 유의할만한 미세누출을 가장 많이 보였으며(p<0.0005) 나머지 3개의 실험군에서는 서로간의 통계학적으로 유의할 만한 차이를 보이지 않았다. 주사전자현미경 관찰 시 제 1군과 제2군의 상아질 접착제가 상아세관으로 침투한 소견을 관찰 할 수 있었다.

• Journal of Periodontal and Implant Science
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• v.30 no.3
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• pp.687-700
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• 2000

Antigen-specific T cell clones were obtained from mice immunized with Fusobacterium nucleatum ATCC 10953(F .nucleatum) and/or Porphyromonas gingi valis 381(P. gingivalis). 10 Balb/c mice per group were immunized with F. nucleatum followed by P. gingivalis, or with P. gingivalis alone by intraperitoneal injection of viable microorganisms. Spleen T cells were isolated and stimulated in vitro with viable P. gingivalis cells to establish P. gingivalisspecific T cell clones. T cell phenotypes and cytokine profiles were determined along with T cell responsiveness to F .nucleatum or P. gingivalis. Serum IgG antibody titers to F. nucleatum or P. gingivalis were also determined by ELISA. All the T cell clones derived from mice immunized with F. nucleatum followed by P. gingivalis demonstrated Th2 subsets, while those from mice immunized with P. gingivalis alone demonstrated Th1 subsets based on the flow cytometric analysis and cytokine profiles, All T cells clones from both groups were cross-reactive to both P. gingivalis and F. nucleatum antigens. Phenotypes of T cell clones were all positive for CD4. Mean post-immune serum IgG antibody levels to F. nucleatum or P . gingivalis were significantly higher than the pre-immune levels(p <0.01, respectively). There were no significant differences in the antibody titers between the two groups. It was concluded that P. gingivalis-specific T cells initially primed by cross-reactive F. nucleatum antigens were polarized to Th2 subsets, while T cells stimulated with P. gingivalis alone maintained the profile of Th1 subset.

• Journal of Periodontal and Implant Science
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• v.24 no.2
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• pp.238-254
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• 1994

Cyclosporine A is an immunosuppressant commonly used for patients receiving organ transplants. Gingival overgrowth is an adverse side-effect seen in about 8-26% of patients taking cyclosporine A which have been shown to increase the DNA synthesis of gingival fibroblast at the concentration of $10^{-9}g/ml$ in vitro. Glycyrrhetinic acid is the active pharmacological ingredients of licorice which exerts steroid-like action and anti-viral activity. Oleanolic acid, which were isolated from Glechoma hederacea, has been shown to act as inhibitors of tumor promotion in vivo and to be less cytotoxic retinoic acid. This study has been performed to evaluate the effects of glycyrrhetinic acid and oleanolic acid on cyclosporine A induced cell activity in vitro. Human gingival fibroblasts were isolated from explant cultures of healthy gingiva of orthodontic patients. Gingival fibroblasts were trypsinized and transferred to the walls of microtest plates. Fibroblasts were cultured in growth medium added $10^{-9}g/ml$ cyclosporineA and $50{\mu}l/ml$ lipopolysaccharides. Cells between the 4th and 6th transfer in culture were used for this study. The morphology of gingival fibroblst were examined by inverted microscope. The effects of cyclosporine A on the time course of DNA sythesis by human gingival fibroblasts were assessed by $[^3H]-thymidine$ uptake assays. Cyclosporine A was found to stimulate DNA synthesis of human gingival fibroblast at a concentration of $10^{-9}g/ml$. In the presence of lipopolysaccharide derived from Fusobacterium nucleatum, addition of cyclosporine A results in reversal of inhibition at the concentration which normally inhibits gingival fibroblast proliferation. The cell acitivities in the presence of glycyrrhetinic acid and oleanolic acid were decreased, and increased cell acitivities by cyclosporine A were decreased by glycyrrhetinic acid and oleanolic acid at the concentration of $200{\mu}g/ml$. These results suggested that the increased cell activities by cyclosporine A modulated by glycyrrhetinic acid and oleanolic acid.

• International Journal of Oral Biology
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• v.37 no.2
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• pp.69-75
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• 2012

Although neutrophils function in both defense and tissue destruction, their defensive roles have rarely been studied in association with periodontitis. We hypothesized that peripheral neutrophils are pre-activated in vivo in periodontitis and that hyperactive neutrophils would show enhanced phagocytic ability as well as an increased production of reactive oxygen species (ROS). Peripheral blood neutrophils from patients with aggressive periodontitis and age/gender-matched healthy subjects (10 pairs) were isolated. The levels of CD11b and CD64 expression on the neutrophils and the level of plasma endotoxin were determined by flow cytometry and a limulus amebocyte lysate test, respectively. In addition, neutrophils were subjected to a flow cytometric phagocytosis assay and luminol-enhanced chemiluminescence for non-opsonized Fusobacterium nucleatum in parallel. The neutrophilsfrom most patients expressed increased levels of both CD11b and CD64. In addition, the plasma from these patients tended to contain a higher level of endotoxin than the healthy controls. In contrast, no differences were found between the two groups with regard to phagocytosis or ROS generation by F. nucleatum. The ability to phagocytose F. nucleatum was found to positively correlate with the ability to produce ROS. In conclusion, peripheral neutrophils from patients with aggressive periodontitis are hyperactive but not hyperreactive to F. nucleatum.

• International Journal of Oral Biology
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• v.41 no.4
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• pp.199-208
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• 2016

The aim of this study was to identify the non-Aggregatibacter actinomycetemcomitans bacteria grown on the tryptic soy-serum-bacitracin-vancomycin (TSBV) medium, an A. actinomycetemcomitans selective medium. A total of 82 unidentified bacterial isolates from the oral cavities of a Korean population were kindly provide by the Korean Collection for Oral Microbiology. All the clinical isolates were grown on TSBV medium and bacterial DNA purified from each isolate was subjected to PCR with universal primers specific for bacterial 16S rRNA genes (16S rDNAs) sequence. The each bacterial 16S rDNA was amplified by PCR and the nucleotide sequences of it was determined by the dideoxynucleotide chain termination method. They were identified by 16S rDNA sequence comparison method at the specie-level. The data showed that Neisseria spp. (42 strains), Fusobacterium spp. (10 strains), Capnocytophaga spp. (8 strains), Propionibacterium acnes (5 strains), Aggregatibacter aprophilus (4 strains), Campylobacter spp. (5 strains), Veillonella dispar (3 strains), Streptococcus sp. (1 strain), Haemophilus parainfluenzae (1 strain), Leptotrichia wadei (1 strain), Morococcus sp./Neisseria sp. (1 strain), and Staphylococcus sp. (1 strain) were identified. These results could be used to develop a new A. actinomycetemcomitans-selective medium which is more effective than the TSBV medium in future studies.

• International Journal of Oral Biology
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• v.39 no.4
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• pp.201-206
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• 2014

This study investigated the antimicrobial activity of methanol extract of mulberry leaf against 16 strains of mutans streptococci and four species of periodontopathogens: Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, and Aggregatibacter actinomycetemcomitans. The antimicrobial activities of the crude extracts or silica gel chromatography fractions of methanol-extracted mulberry leaf were evaluated by determining minimal inhibitory concentrations using an established microdilution method. The cytotoxicity of the extracts of mulberry leaf on KB cells was tested by the methyl thiazolyl tetrazolium assay. Chromatography fraction 12 displayed the most potent antimicrobial activity against all 16 strains of mutans streptococci, P. gingivalis, and P. intermedia. No KB cell cytotoxicity was evident up to $128{\mu}g/ml$ of fraction 12. The methanol extract had no antimicrobial activity against F. nucleatum and A. actinomycetemcomitans. These results suggest chromatography fraction 12 methanol extract of mulberry leaf could be useful in the development of oral hygiene products, such as dentifrice and oral hygiene solution, for the prevention of dental caries.

• Restorative Dentistry and Endodontics
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• v.27 no.5
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• pp.530-534
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• 2002

최근 치근단 와동 형성 시 초음파 또는 음파 기구를 이용하는 것이 보편화되어 있고 여러 측면에서 평가되어져 왔다. 본 연구의 목적은 혐기성 세균을 이용한 미세 누출 모델을 이용하여 초음파와 음파 기구를 사용, 치근단 와동 형성을 하였을 때 두 기구의 사용이 충전된 치근단 와동의 미세 누출에 미치는 영향을 비교 평가하는 것이다. 48개의 단근치의 근관을 crown-down방법을 이용 Profile로 .06 black까지 근관 성형을 시행하고 수직 가압법을 사용하여 gutta-percha와 AH26 sealer를 이용하여 근관 충전을 시행하였다. 각 충전된 시편의 치근단 3mm를 절제하였다. 각 시편은 이미로 4군으로 나누었으며 1군에서는 치근단 와동을 초음파 기구로 형성하였고 2군에서는 음파 기구로 형성하였다. 3군은 음성대조군으로 4군은 양성대조군으로 분류하였다. 형성된 치근단 와동은 공기 분사침을 이용하여 건조 후 super EBA 시멘트로 충전하였으며 Anaerobic chamber에서 Fusobacterium nucleatum (VPI 10197)을 사용한 혐기성 세균 미세누출 모델을 이용하여 30일간 미세 누출을 관찰한 결과, 초음파 기구를 사용한 군에서는 80%에서 음파 기구를 사용한 군에서는 75%에서 미세누출이 나타났으나 통계적으로 유의성 있는 차이는 보이지 않았다.

• Korean Journal of Pharmacognosy
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• v.47 no.1
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• pp.43-48
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• 2016

The aim of the study was performed to examine the anticariogenic potential of purified bee venom (Apis mellifera L., PBV) collected using bee venom collector from cariogenic bacteria, Streptococcus mutans, Streptococcus sanguis, Porphyromonas gingivalis, and Fusobacterium nucleatum. The anticariogenic effect of purified bee venom was evaluated by agar well diffusion method, minimum inhibitory concentraion (MIC), minimum bactericidal concentration (MBC), and postantibiotic effect (PAE). The human lower gingiva epithelial cell cytotoxicity of purified bee venom was also evaluated. Purified bee venom exhibited significant inhibition of bacterial growth of S. mutans, S. sanguis, P. gingivalis, and F. nucleatum with MIC value of 0.68, 0.85, 3.49, and $2.79{\mu}g/ml$, respectively. The MBC value of purified bee venom against S. mutans, S. sanguis, P. gingivalis, and F. nucleatum was 1.34, 1.67, 8.5, and $6.8{\mu}g/ml$. Furthermore, the results of PAE values against S. mutans, S. sanguis, P. gingivalis, and F. nucleatum showed the bacterial effect with 3.3, 3.45, 2.0, and 2.0. The concentration below 1 mg/ml of purified bee venom had no cytotoxicity in the human lower gingiva epithelial cell. These results suggested that purified bee venom have great potenial as anticariogenic agents.