• 한국치위생학회지
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• 제6권4호
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• pp.311-324
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• 2006

The purpose of this investigation was to evaluate of the specificity of Fusobacterium nucleatum subspecies-specific DNA probes using dot blot hybridization. To confirm whether the clinical isolates were F. nucleatum or not, 16S rDNA of them were cloned and sequenced. The sequencing data were used in homology search with database of GenBank. When the homology was above 98% compared with the nucleotide sequence of a certain bacteria, it was judged as the same species with the bacteria. 23 strains of F. nucleatum were isolates from subgingival plaque of periodontitis patient. The clinical isolates of F. nucleatum were classified into 10 groups using phylogenetic analysis of 16S rDNA sequence. F. nucleatum subspecies nucleatum-specific DNA probe Fu4(1.3 kb) reacted with genomic DNAs from 8 type strains of F. nucleatum and it reacted strongly with those from 8 clinical isolates. The Fp4(0.8 kb) reacted with F. nucleatum subsp. polymorphum ATCC 10953 and one clinical isolates. Fv35(1.9 kb) and Fs17(8.2 kb) probes reacted with genomic DNAs from F. nucleatum subsp. vincentii ATCC 49256 and F. nucleatum subsp. fusiform ATCC 51190, respectively. Our results showed that it is not enough to evaluate the specificity of F. nucleatum subspecies-specific DNA probes with only dot blot hybridization. Therefore, Southern blot analysis will be necessary to confirm the specificity of F. nucleatum subspecies-specific DNA probes.

• 미생물학회지
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• 제53권3호
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• pp.219-221
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• 2017

Fusobacterium nucleatum는 그람 음성이면서, 절대 혐기성 및 막대 또는 가는 섬유 모양의 세균이다. F. nucleatum는 사람의 구강 내 정상세균총의 하나이고, 치주질환의 원인인자이면서 다양한 전신질환과도 연관성이 있다. F. nucleatum KCOM 1323 (= ChDC F317) 균주가 사람 치주질환 병소의 치은연하치면세균막에서 분리되었다. F. nucleatum KCOM 1323 균주 유전체 염기서열을 해독하여 보고한다.

• International Journal of Oral Biology
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• 제49권1호
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• pp.1-9
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• 2024

Oral bacterial infections substantially affect the development of various periodontal diseases and oral cancers. However, the molecular mechanisms underlying the association between Fusobacterium nucleatum (F. nucleatum ), a major periodontitis (PT)-associated pathogen, and these diseases require extensive research. Previously, our RNA-sequencing analysis identified a few hundred differentially expressed genes in patients with PT and peri-implantitis (PI) than in healthy controls. Thus, in the present study using oral squamous cell carcinoma (OSCC) cells, we aimed to evaluate the effect of F. nucleatum infection on genes that are differentially regulated in patients with PT and PI. Human oral squamous cell carcinoma cell lines OSC-2O, HSC-4, and HN22 were used. These cells were infected with F. nucleatum at a multiplicity of infection of 100 for 3 hours at 37℃ in 5% CO₂. Gene expression was then measured using reverse-transcription polymerase chain reaction. Among 18 genes tested, the expression of CSF3, an inflammation-related cytokine, was increased by F. nucleatum infection. Additionally, F. nucleatum infection increased the phosphorylation of AKT, p38 MAPK, and JNK in OSC-20 cells. Treatment with p38 MAPK (SB202190) and JNK (SP600125) inhibitors reduced the enhanced CSF3 expression induced by F. nucleatum infection. Overall, this study demonstrated that F. nucleatum promotes CSF3 expression in OSCC cells through p38 MAPK and JNK signaling pathways, suggesting that p38 MAPK and JNK inhibitors may help treat F. nucleatum-related periodontal diseases by suppressing CSF3 expression.

• Journal of Periodontal and Implant Science
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• 제30권3호
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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.

• Restorative Dentistry and Endodontics
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• 제24권1호
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• pp.40-48
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• 1999

This study was designed to examine the specificities of the designed primers for F. nucleatum and F. necrophorum and to compare the PCR results using clinical samples with those of the anaerobic culture method. F. nucleatum and F. necrophorum spp. are frequently isolated in infected root canals, and they are related to periapical diseases. F. nucleatum(VPI 10197) and F. necrophorum(ATCC 25286) were used as references for PCR reaction, and thirty five teeth with one canal and periapical lesion were used. The samples were cultured anaerobically and identified using Rapid ID 32A(BioMerieux Vitek, Inc., France) as biochemical battery. In the GenBank database, species-specific PCR primers(nuc1/nuc2 primers for F. nucleatum and nec1/nec2 primers for F. necrophorum) were designed from the 16S ribosomal DNA(rDNA) sequences of F. nucleatum(accession number M58683) and F. necrophorum(accession number AF044948). PCR procedures of F. nucleatum(VPI 10197) and F. necrophorum (ATCC 25286) were simulated on a computer software. Amplify(v.1.2${\beta}$ for Macintosh). 820 bps and 817 bps of nucleotides were expected, respectively. Using extracted DNAs with QiaAmp tissue kit(Qiagen co., Germany), PCR was done. The results were as follows : 1. The nuc1/nuc2 primers produced an amplicon of 820 bps and the nec1/nec2 primers produced an amplicon of 817 bps. 2. The nuc1/nuc2 primers and the nec1/nec2 primers were specific and did not react with species other than the designated ones(i.e. nuc1/nuc2 primers did not produce amplicons for F. necrophorum, and vice versa.). And the PCR products of Porphyromonas endodontalis(ATCC 35406), Porphyromonas gingivalis(ATCC 33277), Prevotella intermedia(ATCC 25611), and Prevotella nigrescens(ATCC 33563), frequently isolated in infected root canals and periapical lesions, were not amplified by the primers specific for Fusobacterium nucleatum and Fusobacterium necrophorum. 3. This method utilizing PCR could detect F. nucleatum and F. necrophorum in clinical samples, while anaerobic culture method could detect neither.

• Journal of Periodontal and Implant Science
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• 제33권2호
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• pp.149-158
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• 2003

The purpose of this study was to isolate and characterize the Fusohacrerium nucleatum (F. nucleatum) from subgingival plaque in Korean periodontitis patients. The subgingival plaque samples of periodontitis patient were collected with sterilized paper point. The paper point was put into reduced transfer medium and then immediately transferred to laboratory. The subgingival samples were diluted by 10,000 folds and plated on F. nucleatum-selective media agar plate. The plates were incubated at 37$^{\circ}C$ in an anaerobic chamber for 3 days. The violet-colored colonies were selected and subjected to further verification whether those are F. nucleatum or not. For further confirmation, 16S rRNA genes (rDNA) were cloned from each of bacterial clones and determined sequence of 16S rDNA. In this study, we found 17 distinct clinical isolates of F. nucleatum from subgingival plaque. The clinical isolates will be a useful in various studies in periodontology.

• Journal of Oral Medicine and Pain
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• 제34권4호
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• pp.353-362
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• 2009

본 연구는 편백 피톤치드에 의해 사멸되지 않는 구강 상주균을 분리하고, 이 분리된 세균이 구강 병인균에 대하여 어떠한 영향을 미치는지를 관찰함으로써 편백 피톤치드에 구강 내 세균에 대한 지속적인 이차적 효과를 구명한 실험적 연구이다. 이에 건강한 사람의 타액에 1% 피톤치드를 첨가하여 최종적으로 생존해 있는 200개의 구강 상주균을 확인하여 이를 분석한 후 이들이 치주질환과 입냄새의 중요한 원인균인 F. nucleatum에 대한 억제효과를 관찰하여 다음과 같은 결론을 얻었다. 1. 선택된 200개의 잔존 세균 중, 70개(35.0%)가 F. nucleatum을 억제하였다. 2. F. nucleatum을 억제하는 70개의 잔존 세균 중, Streptococcus salivarius가 41.3%(45/109), Streptococcus sanguinis가 28%.(7/25), Streptococcus mitis가 20%(3/15), Streptococcus parasanguinis가 33.3%(3/9), Streptococcus alactolyticus가 100%(8/8), Streptococcus vestibularis가 28.6%(2/7), Streptococcus sp.가 50%(2/4)로 나타났다. 결론적으로 피톤치드 처리 후의 잔존 세균이 F. nucleatum을 억제함으로써 피톤치드가 구강 건강, 특히 치주질환을 예방하거나 치료하는데 큰 역할을 할 수 있을 것으로 생각하며, 구강 상주균을 건강하게 유지시킨 채로 병인균 만을 억제시킨다는 차원에서 향후 구강내 유익균의 배양가능성을 실용화 할 수 있다고 생각한다.

• 치위생과학회지
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• 제9권2호
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• pp.249-255
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• 2009

본 연구는 성인성 치주염 환자의 치은연하 치면세균막을 총 60개 치아에서 채취하여 F. nucleatum과 A. actinomycetemcomitans의 동정을 위해 세균배양법, single PCR법 및 mutliplex PCR법을 실시하였고, 세균 동정법간의 비교를 통해 다음과 같은 결과를 얻었다. 1. F. nucleatum과 A. actinomycetemcomitans의 동정을 위해 세균배양법, single PCR 및 multiplex PCR을 실시한 결과 F. nucleatum은 각각 12개(20.0%), 45개(75.0%), 43개(71.7%) 치아에서 양성반응을 보였지만, A. actinomycetemcomitans는 각각 0개(0.0%), 4개(6.7%), 1개(1.7%) 치아에서 양성반응이 나타났다. 2. F. nucleatum은 세균배양법에 비해 single PCR법 및 multiplex PCR법에서 높은 검출 빈도를 보여 좀 더 효율적인 세균 동정법으로 생각되었지만, 통계적으로는 유의한 차이가 없었다. 3. A. actinomycetemcomitans는 세균배양법에서 전혀 검출되지 않아 통계적으로 검정할 수 없었고, 세균 동정법간의 비교도 어려웠다. 4. F. nucleatum과 A. actinomycetemcomitans의 동정을 위한 single PCR법과 multiplex PCR법 간의 비교에서 두 세균 모두 검출 빈도에 있어서는 큰 차이를 보이지 않았지만, 통계적으로는 유의한 차이를 보였다.

• Journal of Microbiology
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• 제43권4호
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• pp.331-336
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• 2005

The objective of this study was to assess the strain-specificity of a DNA probe, Fu12, for Fusobacterium nucleatum subsp. nucleatum ATCC $25586^T$ (F. nucleatum ATCC $25586^T$), and to develop sets of strain-specific polymerase chain reaction (PCR) primers. Strain-specificity was tested against 16 strains of F. nucleatum and 3 strains of distinct Fusobacterium species. Southern blot hybridization revealed that the Fu12 reacted exclusively with the HindIII-digested genomic DNA of F. nucleatum ATCC $25586^T$. The results of PCR revealed that three pairs of PCR primers, based on the nucleotide sequence of Fu12, generated the strain-specific amplicons from F. nucleatum ATCC $25586^T$. These results suggest that the DNA probe Fu12 and the three pairs of PCR primers could be useful in the identification of F. nucleatum ATCC $25586^T$, especially with regard to the determination of the authenticity of the strain.

• International Journal of Oral Biology
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• 제30권2호
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• pp.59-63
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• 2005

Postantibiotic effects (PAE) refer to suppression of the bacterial growth following limited periods of exposure to an antibiotic and subsequent to the removal of the antibiotic agent. Fusobacterium nucleatum and Porphyromonas gingivalis are Gram-negative anaerobic bacteria associated with several periodontal diseases. In this study, postantibiotic effects (PAE), postantibiotic sub-MIC effect (PA SME) and sub-MIC effect (SME) of antibiotics on F. nucleatum ATCC 25586 and P. gingivalis W50 were investigated. The PAE was induced by 10X the MIC of antibiotic and antibiotic was eliminated by washing. The PA SMEs were studied by addition of 0.1, 0.2 and 0.3X MICs during the postantibiotic phase of the bacteria, and the SMEs were studied by exposition of the bacteria to antibiotic at the sub-MICs only. Amoxicillin, doxycycline and tetracycline induced PAE for F. nucleatum ATCC 25586 and P. gingivalis W50. But metronidazole and penicillin induced PAE for only F. nucleatum ATCC 25586. Metronidazole and doxycycline induced PA SME and SME for both species of anaerobic bacteria used in this study. The PA SME values for both strains were substantially longer than the SME values. The present study showed the existence of PAE, PA SME and SME for various antibiotics against F. nucleatum ATCC 25586 and P. gingivalis W50.