• Journal of Periodontal and Implant Science
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• 제32권4호
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• pp.827-836
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• 2002

The present study was performed to evaluate how cellular and humoral immune responses were perturbed by immunization of mixed periodontal bacterial biofilms. Each group of mice was immunizared with 1) Poqhyromonas gingivalis (P. gingivaliis) grown as a planktonic culture, 2) Fusobacterium nucleatum (F. nucleatum), 3) P. gingivalis grown as a biofilm, or 4) mixed P. gingivalis plus F. nucleatum grown as a biofilm culture, respectively. Immune mouse sera were collected from each mouse. Spleens were harvested to isolate T cells and consequently stimulated with antigen presenting cells and P. gingivalis whole cell antigen to establish P. gingivalis-specific T cell lines. There were no significant differences in the mean anti- gingivalis IgG antibody titers among mouse groups. Immunization of mice with pure P. gingivalis biofilm or mixed P gingivalis plus F. nucleatum biofilm resulted in significant reduction o f antibody avidity and opsonophagocytois function. INF-$\gamma$production by P. gingivalis-specific T cell lines was also substantially recluced in mouse groups immunized with the biofilm. It was concluded that P. gingivalis biofilm perturbs the cellular and humoral immune responses in periodontal disease.

• Journal of Microbiology and Biotechnology
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• 제14권1호
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• pp.110-115
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• 2004

This study was undertaken to develop PCR primers for the simultaneous detection of Fusobacterium nucleatum and Actinobacillus actinomycetemcomitans, using two species-specific reverse primers in combination with a single conserved forward primer. These primers target the variable and conserved regions of the 16S rDNA. The primer specificity was tested against (i) four F. nucleatum and three A. actinomycetemcomitans strains and (ii) seven representatives of the different species of oral bacteria. The primer sensitivity was determined by testing serial dilutions of the purified genomic DNA of F. nucleatum and A. actinomycetemcomitans. The data indicate that species-specific amplicons could be obtained for all the F. nucleatum and A. actinomycetemcomitans strains tested, which were not found in the seven other species. The multiplex PCR could detect as little as 4 fg of chromosomal DNA of F. nucleatum and A. actinomycetemcomitans simultaneously. These findings suggest that these PCR primers are highly sensitive and are suitable for applications in epidemiological studies, diagnosis, and monitoring F. nucleatum and A. actinomycetemcomitans after the treatment of periodontitis.

• Journal of Periodontal and Implant Science
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• 제35권2호
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• pp.401-411
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• 2005

The aim of this study was to determine the minimal inhibitory concentration(MIC) of cefixime, which is a 3rd generation of cefalosporin, against 6 species of putative periodontopathogens; Fusobacterium nucleatum, Actinobacillus actinomycetemcomitans, Prevotella intermedia, Prevotella nigrescens, Tannerella forsythia and Porphyromonas gingivalis. The efficacy of cefixime was examined by comparing it with that of several antibiotics(amoxicillin, $Augmentin^{(R)}$ ciprofloxacin, metronidazole, and tetracycline), which were used as the control. The MIC was measured using a microdilution method. The MIC of cefixime against the putative periodotopathogens, as a single use regimen, was relatively lower than that of the other antibiotics. The MIC of cefixime/metronidazole against P. intermedia ChDC KB14, P. nigrescens ChDC KB50, F. nucleatum ChDC PV-F37, F. nucleatum ChDC F130, and F. nucleatum ChDC F175, as a simultaneous regimen, was lower than that of the other antibiotics. The concentration of cefixime in the crevicular fluid of volunteers who received 250mg every 12 hours for 3 days was $9{\mu}g/ml$ after 9 hours. In conclusion, cefixime showed good antimicrobial activity in a single treatment or as a combined therapy with amoxicillin, $Augmentin^{(R)}$ or metronidazole against 6 periodontopathogens.

• Journal of Periodontal and Implant Science
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• 제31권4호
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• pp.661-668
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• 2001

P. gingivalis를 단독면역하거나 또는 Fusobacterium nucleatum 선면역 후 P. gingivalis 항혈청을 각각 얻어냈다. 두 종류의 항혈청이 P. gingivalis biofilm을 침투해 들어가는 능력을 confocal laser scanning microscope를 이용하여 비교 감증하였다. 항혈청의 P. gingivalis에 대한 avidity index도 측정하였다. 결과적으로 F. nucleatum의 선면역은 P. gingivalis 특이 항혈청에 대해 세균성 biofilm의 침투능력을 저하시키고, 동일한 세균에 대한 avidity도 감소시켰다.

• International Journal of Oral Biology
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• 제35권4호
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• pp.197-202
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• 2010

Most oral microorganisms exist as biofilms which initiate formation via the attachment of an early colonizer to host proteins on the tooth surface. Fusobacterium nucleatum act as a bridge between early and late colonizers. Dental biofilms eventually comprise dental pathogens such as Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia. To evaluate the effects of mutual interactions between oral bacteria on the growth of biofilms, periodontopathogens were co-cultured with a $0.4\;{\mu}m$ barrier. Streptococcus gordonii inhibited the growth of F. nucleatum and periodontopathogens. However, F. nucleatum, P. gingivalis and T. denticola activated the growth of other bacteria. A co-culture system of early and late colonizers could be a useful tool to further understand bacterial interactions during the development of dental biofilm.

• International Journal of Oral Biology
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• 제39권2호
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• pp.115-120
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• 2014

Minimal inhibitory concentration (MIC) is the lowest concentration of antibiotics that inhibits the visible growth of a microorganism. It has been reported that sub-MIC of antibiotics may result in morphological alterations along with biochemical and physiological changes in bacteria. The purpose of this study was to examine morphological changes of periodontal pathogens after treatment with sub-MIC antibiotics. Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, and Porphyromonas gingivalis were used in this study. The MIC for amoxicillin, doxycycline, metronidazole, penicillin, and tetracycline were determined by broth dilution method. The bacterial morphology was observed with bright field microscope after incubating with sub-MIC antibiotics. The length of A. actinomycetemcomitans and F. nucleatum were increased after incubation with metronidazole; penicillin and amoxicillin. P. gingivalis were increased after incubating with metronidazole and penicillin. However, F. nucleatum showed decreased length after incubation with doxycycline and tetracycline. In this study, we observed that sub-MIC antibiotics can affect the morphology of periodontal pathogens.

• International Journal of Oral Biology
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• 제43권4호
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• pp.193-200
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• 2018

The purpose of this study is to determine if natural extracts could be used as an additive in oral health food made with Weissella cibaria CMU (oraCMU). Natural extracts of green tea, mulberry leaf, licorice, and propolis, which are reported to have antimicrobial activities, were selected and used in this study. The minimum inhibitory concentrations (MIC) of extracts on periodontal pathogens such as Fusobacterium nucleatum and Porphyromonas gingivalis and their synergy effects with oraCMU by the fractional inhibitory concentrations methods were measured. From the results obtained, all the extracts showed no effect on the growth of oraCMU. Green tea extract showed the best antibacterial activity with MIC of 1.8 mg/ml against both F. nucleatum and P. gingivalis. In addition, green tea extract had a synergistic effect with oraCMU against F. nucleatum. Therefore, these results suggested that green tea extract is available as an additive in oral health food made with oraCMU.

• Journal of Oral Medicine and Pain
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• 제30권1호
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• pp.1-14
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• 2005

성인의 양호한 구강위생상태 유지에 있어 주요 저해인자로 작용하며, 국소적으로 세균요인 측면에서 통상의 합성 항생제 또는 항균성을 가지는 화학물질에 낮은 감수성을 보이는 수종의 그람음성의 혐기성 세균들에 의해 유발되는 구취와 치주질환에 대한 세균요인 관리 측면에서의 효율적 치료를 위한 천연 항균성 구강함수제의 개발에 필요한 기초자료를 얻고자, 구취와 치주질환의 유발과 관련된 대표적 구강내 그람음성의 혐기성 세균들인 P. intermedia속, P. gingivalis속 및 F. nucleatum 속 가운데 수종의 균주들에, 천연물의 일종인 은으로부터 제조된 콜로이드상 은 용액을 농도(10 ppm, 30 ppm, 50 ppm, 80 ppm)를 달리 적용한 후, 혐기적 상태에서의 경과시간(24 시간, 48 시간, 72 시간)에 따른 증식억제율과 치사율을 측정하여, 최소증식억제농도(minimal inhibitory concentration), 최소세균치사농도(minimal bactericidal concentration) 및 항균효과의 양상을 분석, 평가하여 다음과 같은 결과를 얻었다. 1. 콜로이드상 은의 실험대상 세균에 대한 최소증식억제농도는 P. intermedia속의 경우 30 ppm, P. gingivalis속의 경우에는 실험 균주에 따라 10 ppm 또는 30 ppm, 그리고 F. nucleatum속의 경우에는 실험 균주에 따라 30 ppm, 50 ppm 또는 80 ppm이었다. 2. 콜로이드상 은의 실험대상 세균에 대한 최소세균치사농도는 P. intermedia속의 경우 30 ppm, P. gingivalis속의 경우에는 실험 균주에 따라 30 ppm 또는 50 ppm, 그리고 F. nucleatum속의 경우에는 실험 균주에 따라 30 ppm 또는 80 ppm이었다. 3. 콜로이드상 은의 실험대상 균주에 대한 항균효과는 동일한 농도에서도 실험대상 균주에 따라 정균효과나 살균효과, 또는 양자 모두에 기인하였다. 4. 콜로이드상 은이 농도별로 적용된 후 경과시간에 따른 실험대상 세균의 증식에 미치는 효과에 있어서 P. intermedia속 균주의 경우 콜로이드상 은이 30 ppm$\sim$50 ppm의 농도로 48 시간$\sim$72 시간, P. gingivalis속 균주의 경우 10 ppm$\sim$30 ppm의 농도로 24 시간$\sim$48 시간, 그리고 F. nucleatum속 균주의 경우에는 30 ppm의 농도로 24 시간 동안 적용된 경우 대조군에 비해 균주에 따라 현저하게 또는 보통수준으로 증식이 억제되었다.

• 치위생과학회지
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• 제15권2호
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• pp.209-219
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• 2015

치주질환과 관련된 세균인 S. gordonii, F. nucleatum 및 P. gingivalis의 상호작용이 군집 형성에 미치는 영향을 알아보고자 trypticase soy hemin menadione broth에 단독 및 열처리한 사균과 혼합 분주하여 혐기성 균배양조를 통해 $37^{\circ}C$ $CO_2$ 배양기에서 anearobic gas pack 하에 7일간 배양하였다. 군집 형성 정도를 확인하기 위해 흡광도를 측정하였으며, 군집 구조 및 형태를 확인하기 위해 주사전자현미경으로 관찰하였다. P. gingivalis의 병원성에 미치는 영향을 확인하기 위해 real-time RT-PCR를 통해 gingipain인 HRgpA를 생성하는 rgpA 유전자에 대한 발현 분석을 시행하여 다음과 같은 결론을 얻었다. S. gordonii와 P. gingivalis의 군집 형성은 다른 사균들에 의해 증가하였다. F. nucleatum의 경우 P. gingivalis 사균에 의해 증가하는 양상을 보였으나 S. gordonii 사균에 의해서는 군집 형성이 감소되었다. 따라서 본 실험에 사용된 균주는 군집 형성 시 상호작용 인자뿐 아니라 세균 입자 그 자체 등을 통해서도 서로 영향을 주는 것으로 생각된다.

• International Journal of Oral Biology
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• 제41권4호
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• pp.237-242
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• 2016

Interleukin-1b ($IL-1{\beta}$), a proinflammatory cytokine, regulates the innate immune responses against bacterial infection. Mature $IL-1{\beta}$ is produced from $pro-IL-1{\beta}$ by activated caspase-1, which in turn is activated by the inflammasome complex formation. In this study, we compared the inflammasome mRNA expression induced by S. sanguinis, S. oralis, F. nucleatum and P. intermedia. Among the tested bacteria, S. sanguinis induced the highest $IL-1{\beta}$ secretion. S. oralis, F. nucleatum and P. intermedia induced very weak $IL-1{\beta}$ secretion. S. sanguinis mostly induced the NLRP3 mRNA expressions. Although F. nucleatum did not induce high $IL-1{\beta}$ secretion, it induced high expression levels of AIM2, NLRP2, and NLRP3. No specific inflammasomes were induced by S. oralis and P.intermedia. Studying the inflammasome complex activation induced by oral bacteria may thus enhance our understanding of the pathogenesis of oral diseases.