• Journal of Periodontal and Implant Science
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• 제30권2호
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• pp.419-429
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• 2000

Prevotella intermedia has been implicated as a potent pathogen in many kinds of periodontal, pulpal and periapical diseases. However, it has been isolated from periodontally healthy adults and from edentulous children as well. The intraspecies heterogeneity of Prevotella intermedia has been demonstrated in early studies and finally Shah & Gharbia confirmed the existence of 2 DNA homology groups and proposed dividing Prevotella intermedia into 2 species, Prevotella intermedia and Prevotella nigrescens. This study was designed to examine the frequency of Prevotella intermedia and Prevotella nigrescens in diseased periodontal pockets and healthy gingival sulcus of Korean people by PCR based on 16s ribosomal DNA sequence. One hundred adults who had adult periodontitis but not taken any periodontal treatment or antibiotics during previous 6 months and 50 adults who had healthy periodontal tissue were selected for this study. The sulcular fluid was collected into VMGA by sterilized paper point and diluted to 1,000 times in anaerobic chamber. $100{\mu}{\ell}$ of sample was cultured in $37^{\circ}C$ for 10 days. Among the bacterial colonies, BPB were selected and cultured in BHI broth and then Prevotella intermedia was identified through Gram staining and biochemical test. Identified Prevotella intermedia was cultured again and centrifuged. DNA was extracted from the pellet using several reagents. PCR was performed by previously designed primer. The results were followed. 1. BPB were isolated from 39 of 100 samples of diseased periodontal pockets(39%). 2. Prevotella intermedia was identified from 24 of 39 BPB samples. 3. Among 24 Prevotella intermedia, 21 were confirmed as Prevotella inter - media(87.5) and 2 were confirmed as Prevotella nigrescens(8.33%). 4. BPB were isolated from 9 of 50 samples of periodontally healthy patients. Among them only two were identified as Prevotella intermedia, that is, one was confirmed as Prevotella intermedia and the other was Prevotella nigrescens.

• Restorative Dentistry and Endodontics
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• 제23권2호
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• pp.550-560
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• 1998

Prevotella intermedia has been known as one of the important bacterial species involved in the endodontic infections and various periodontal diseases. Polyphosphate has been widely used to prevent decomposition of food and known to have an inhibitory effect on the growth of gram positive bacteria. The purpose of this study was to evaluate the effect of poly phosphate on the growth of Prevotella intermedia, a gram negative bacterium. Prevotella intermedia G8GK3(ATCC 49046) was grown in the presence of polyphosphates with different chain lengths. Inhibitory effect of each polyphosphate, which was added at the beginning or at the early exponential growth phase of Prevotella intermedia, was determined by measuring optical density of the bacterial cells at 540nm, viable cells and lysis of Prevotella intermedia. The results from this study were as follows : 1. Poly phosphate inhibited the growth of Prevotella intermedia. 2. The minimum inhibitory concentration(MIC) of poly phosphate appeared to be 0.05%. 3. Polyphosphates with chain lengths of 5 and 65 demonstrated the greatest inhibitory effect on the growth of Prevotella intermedia. 4. Polyphosphate was bactericidal to Prevotella intermedia, demonstrating the growth inhibition of the bacterium. 5. Polyphosphate induced lysis of Prevotella intermedia. The overall results suggest that polyphosphate has a bactericidal effect on Prevotella intermedia, causing the lysis of the bacterium.

• Journal of Periodontal and Implant Science
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• 제39권2호
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• pp.177-184
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• 2009

Purpose: Interleukin-8 (IL-8) is an important mediator of immune and inflammatory reactions and is produced by a variety of different cell types. This study was undertaken to investigate the effects of lipopolysaccharides (LPSs) from Prevotella intermedia and Prevotella nigrescens, the major causes of inflammatory periodontal disease, on the production of IL-8 and the expression of IL-8 mRNA in differentiated THP-1 cells, a human monocytic cell line. Methods: LPSs from P. intermedia ATCC 25611 and P. nigrescens ATCC 33563 were prepared by the standard hot phenol-water method. THP-1 cells were incubated in the medium supplemented with phorbol myristate acetate to induce differentiation into macrophage-like cells. Results: We found that LPS preparations from P. intermedia and P. nigrescens can induce IL-8 mRNA expression and stimulate the release of IL-8 in differentiated THP-1 cells without additional stimuli. Conclusions: There are no previous reports of the ability of P. intermedia and P. nigrescens LPS to stimulate the release of IL-8, and the present study clearly shows, for the first time, that LPSs from P. intermedia and P. nigrescens fully induced IL-8 mRNA expression and IL-8 production in differentiated human monocytic cell line THP-1. The ability of P. intermedia and P. nigrescens LPS to promote the production of IL-8 may be important in the pathogenesis of inflammatory periodontal disease.

• Journal of Periodontal and Implant Science
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• 제34권2호
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• pp.461-474
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• 2004

The purpose of this study was to assess some biological activities of lipopolysaccharides (LPSs) from P. intermedia and P. nigrescens. LPS was prepared by the standard hot phenol-water method. NO production was assayed by measuring the accumulation of nitrite in culture supernatants. $TNF-{\alpha}$ production was determined by enzyme-linked immunosorbent assay. Western blot analysis of iNOS and analysis of reverse transcription (RT)-PCR products were carried out. LPS from P. intermedia demonstrated higher KDO content than those from two stains of P. nigrescens. LPSs from P. intermedia and P. nigrescens were mitogenic for spleen cells of BALB/C mouse. The present study clearly shows that LPSs from P. intermedia and P. nigrescens fully induced iNOS expression and NO production in RAW264.7 cells in the absence of other stimuli. Moreover, LPSs from P. intermedia and P. nigrescens clearly induced $TNF-{\alpha}$ production in RAW264.7 cells. The biological activities of LPS from P. intermedia was found to be comparable to those of P. nigrescens LPS. The ability of LPSs from P. intermedia and P. nigrescens to promote the production of NO and $TNF-{\alpha}$ may be important in the pathogenesis of inflammatory periodontal disease.

• Journal of Periodontal and Implant Science
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• 제36권1호
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• pp.155-165
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• 2006

The results of this study confirm that the availability of hemin influences the expression of selected membrane proteins of Porphyromonas gingivalis, Prevotella intermedia, and Prevotella nigrescens. A 30 kDa (heated 24 kDa) hemin-binding protein whose expression is hemin regulated was identified and purified in P. gingivalis. A strong hemin-binding function was found by LDS-PAGE and TMBZ staining when P. gingivalis cells were grown under hemin-limited conditions. A 50 kDa cell envelope associated protein, whose expression is hemin regulated, is considered to be a putative hemin binding protein from P. intermedia and P. nigrescens, respectively. N-terminal amino acid sequence analysis of CNBr-digested 24 kDa hemin binding protein from P. gingivalis revealed that this protein belongs to a new, so far undescribed hemin-binding class of proteins. N-terminal amino acid sequence of a 50 kDa putative hemin binding protein from P. intermedia was identical with Enolase from Streptococcus intermedia. Work is in progress to further characterize the molecular structure of these proteins.

• 한국미생물·생명공학회지
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• 제31권2호
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• pp.205-209
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• 2003

Recently, we introduced a new method for rapid screening of bacterial species- or subspecies-specific DNA probes, named “inverted dot blot hybridization screening method”. We then applied this method to develop species- or strain- specific DNA probes for Prevotella intermedia and Prevotella nigrescens. In those studies, among 96 candidate DNA probes which were screened by the new method, 5 probes were confirmed as being putatively strain-specific : 3 probes for P. nigrescens 9336 (ATCC 33563), one for each p. intermedia ATCC 25611 and one for P. nigrescens G8-9K-3 (ATCC 49046). In the present study, we evaluated by Southern blot analysis a DNA probe Pi29-L, one of the 96 candidate probes described above, whether it is specific for the strain ATCC 25611 off. intermedia. Our data show that the probe Pi29-L is potentially P. intermedia ATCC 25611-specific, which can be useful for the detection and identification of the strain, particularly in maintenance of the strain.

• 미생물학회지
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• 제53권3호
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• pp.222-224
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• 2017

Prevotella intermedia는 그람 음성이면서, 절대 혐기성, 비스포아형성 및 운동성 간균이다. P. intermedia는 사람의 구강내 정상세균총의 하나이며, 치주질환, 임신성 치은염, 급성괴사성궤사성 치은염, 치근관 감염, 및 류마티성 관절염과 관련이 있다. P. intermedia KCOM 1107 (= ChDC KB29) 균주가 사람 치은염 병소 치은연하 치면세균막에서 분리되었다. P. intermedia KCOM 1107 균주 유전체 염기서열을 해독하여 보고한다.

• International Journal of Oral Biology
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• 제36권2호
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• pp.79-82
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• 2011

The aim of this study was to develop Prevotella intermedia ATCC 49046-specific PCR primers designed based on the nucleotide sequence of a DNA probe Pig28. The strainspecificity of the PCR primers, Pig28-F1/Pig28-R1, was confirmed with 9 strains of P. intermedia and 25 strains (15 species) of Prevotella species. The detection limit of the PCR primers was 2 pg of the purified genomic DNA of P. intermedia ATCC 49046. These PCR primers were found to be useful for identifying P. intermedia ATCC 49046, particularly for determining the authenticity of the strain.

• Journal of Periodontal and Implant Science
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• 제30권4호
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• pp.737-746
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• 2000

본 연구는 hemin이 치주질환 주요 병인균주 중의 하나인 Prevotella intermedia의 성장 및 세포막 단백질의 발현에 미치는 영향을 규명하고, hemin 결합에 관여하는 것으로 추정되는 단백질의 순수분리 및 특성 분석을 위해 수행 되었다. 본 연구의 결과에 의하면, hemin은 P.intermedia의 성장 및 세포막 단백질의 발현에 영향을 미쳐, hemin이 고갈된 조건에서 균주의 성장이 현저히 억제되었으며, 약 50 kDa의 세포막 단백질이 현저히 강화되어 발현되었다. 본 연구에서는 hemin 결합에 관여하는 것으로 추정되는 이 50 kDa의 세포막 단백질을 순수분리하였으며, N'-terminal 아미노산 분석을 수행한 결과 이 단백질은 Streptococcus inter - medius의 Enolase와 아미노산 서열 및 분자량이 일치하였다. 한편, 이 단백질에는 disulfidebond가 존재하지 않았다. 본 연구는 P.intermedia에서의 porphyrin 생리 및 hemin 획득기전을 밝히는데 있어 중요한 의의가 있으리라 사료된다.

• 대한화장품학회지
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• 제37권1호
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• pp.67-73
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• 2011

치아의 세균들은 잇몸에서 염증반응을 일으켜서 치은염과 치주염같은 잇몸 질환의 원인이 된다. 따라서 잇몸질환의 예방과 치료를 위해서는 치아 세균에 의한 염증반응을 조절하는 것이 가장 효과적인 방법이다. 현재 대부분의 구강 관리 제품들은 살균제를 이용하여 구강 세균을 제거하는 방법을 사용하고 있다. 하지만 최근의 연구들은 심지어 열처리로 사멸된 구강 세균도 염증반응을 일으킬 수 있다는 사실을 보고하고 있다. 따라서 보다 효과적인 잇몸 염증반응억제를 위해서는 살균제를 이용한 방법에 대한 개선이 필요하다. 또한 아직까지 구강 세균에 의한 잇몸 염증반응의 기작과 효과적인 천연 염증 억제 물질들은 보고되어 있지 않다. 본 연구에서는 대표적인 치은염, 치주염 유발세균인 Prevotella intermedia와 인간의 잇몸상피세포를 이용하여, 실제로 잇몸에서 일어나는 염증반응의 기작을 연구하고, 이를 통해 효과적인 천연 잇몸 염증 완화 물질을 도출하려고 하였다. 실험 결과, Prevotella intermedia는 잇몸상피세포를 자극하여 염증매개인자인 IL-8을 분비하게 함으로써 잇몸 염증반응을 개시하였다. 또한 Prevotella intermedia에 의한 잇몸 염증반응은 기전적으로 COX-2, AP-1, TNF-${\alpha}$와 연관되어 있었으며, 녹차추출물은 Prevotella intermedia에 의한 잇몸 염증반응을 효과적으로 억제할 수 있음을 확인하였다. 따라서 본 연구는 치아 세균에 의한 잇몸 염증반응의 기전 연구를 통해서 효과적인 천연 잇몸질환 개선 물질을 도출했다는 점에서 중요한 의미를 가진다.