• Journal of Periodontal and Implant Science
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• 제38권3호
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• pp.511-520
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• 2008

Purpose: Diabetes mellitus is a clinically and genetically heterogeneous group of metabolic disorders manifested by abnormally high levels of glucose in the blood. Mounting evidence demonstrates that diabetes is a risk factor for gingivitis and periodontitis. The circulating mononuclear phagocytes in diabetic patients with hyperglycemia are chronically exposed to high level of serum glucose. Thus, this study attempted to determine the effect of pre-exposure of monocytes and macrophages to high concentration of glucose on lipopolysaccharide (LPS)-induced production of pro-inflammatory mediators. Material and Methods: For this purpose, cells were cultured in medium containing normal (5 mM) or high glucose (25 mM) for 4-5 weeks before treatment for 24 h with LPS. LPS was highly purified from Porphyromonas gingivalis or Prevotella intermedia by phenol extraction. Result: Results showed that prolonged pre-exposure of cells to high glucose markedly increased LPS-stimulated NO secretion when compared to normal glucose. In addition to NO, high glucose also augmented LPS-stimulated IL-6, IL-8, and TNF-$\alpha$ secretion after cells were exposed to high glucose for 4 weeks. Conclusion: The present study demonstrates that pre-exposure of mononuclear phagocytes with high glucose augments LPS-stimulated production of pro-inflammatory mediators. These findings may explain why periodontal tissue destruction in diabetic patients is more severe than that in non-diabetic individuals.

• International Journal of Oral Biology
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• 제46권3호
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• pp.111-118
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• 2021

Periodontitis and periimplantitis are caused as a result of dental biofilm formation. This biofilm is composed of multiple species of pathogens. Therefore, controlling biofilm formation is critical for disease prevention. To inhibit biofilm formation, sugars can be used to interrupt lectin-involving interactions between bacteria or between bacteria and a host. In this study, we evaluated the effect of D-Arabinose on biofilm formation of putative periodontal pathogens as well as the quorum sensing activity and whole protein profiles of the pathogens. Crystal violet staining, confocal laser scanning microscopy, and scanning electron microscopy revealed that D-Arabinose inhibited biofilm formation of Porphyromonas gingivalis, Fusobacterium nucleatum, and Tannerella forsythia. D-Arabinose also significantly inhibited the activity of autoinducer 2 of F. nucleatum and the expression of representative bacterial virulence genes. Furthermore, D-Arabinose treatment altered the expression of some bacterial proteins. These results demonstrate that D-Arabinose can be used as an antibiofilm agent for the prevention of periodontal infections.

• 생약학회지
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• 제52권1호
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• pp.41-48
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• 2021

This research was conducted to investigate the antibacterial and anti-inflammatory effects of MeOH Ex. of Salvia miltiorrhiza (MESM) against oral pathogenic bacteria. Minimum inhibitory concentration (MIC), removal effect of biofilm produced by Streptococcus mutans, effect of gene expression of proinflammatory cytokines and effect of production of proinflammatory cytokine of MESM were tested. MESM showed moderated antibacterial activity against oral pathogenic bacteria. About 89±8% of biofilms produced by S. mutans were removed by MESM at a concentration of 1 mg/mL. Gene expression of IL-1β and TNF-α induced by Porphyromonas gingivalis were 8~9 folds reduced by MESM. Gene expression of IL-8 induced by Fusobacterium nucelatum were 12 folds reduced by MESM. Production of IL-1β, TNF-α and IL-8 were significantly suppressed by MESM. Conclusively, MESM showed potent antibacterial and anti-inflammatory effect against oral pathogenic bacteria.

• International Journal of Oral Biology
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• 제47권2호
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• pp.25-31
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• 2022

Lysophosphatidic acid (LPA) is a bioactive lipid messenger involved in the pathogenesis of chronic inflammation and various diseases. Recent studies have shown an association between periodontitis and neuroinflammatory diseases such as Alzheimer's disease, stroke, and multiple sclerosis. However, the mechanistic relationship between periodontitis and neuroinflammatory diseases remains unclear. The current study found that lysophosphatidic acid receptors 1 (LPAR1) and 6 (LPAR6) exhibited increased expression in primary microglia and astrocytes. The primary astrocytes were then treated using medium conditioned to mimic periodontitis through addition of Porphyromonas gingivalis lipopolysaccharides, and an increased nitric oxide (NO) production was observed. Application of conditioned medium from human periodontal ligament stem cells with or without LPAR1 knockdown showed a decrease in the production of NO and expression of inducible nitric oxide synthase and interleukin 1 beta. These findings may contribute to our understanding of the mechanistic link between periodontitis and neuroinflammatory diseases.

• 한국치위생학회지
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• 제24권2호
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• pp.131-139
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• 2024

Objectives: Oral bacterial samples included subgingival, supragingival, and saliva plaques. As the diversity and number of microorganisms deffer depending on the area of the oral cavity and the method used, an appropriate and reliable collection method is important. The present study investigated oral bacterial sampling methods. Methods: Supragingival dental plaque was collected from the buccal and lingual tooth surfaces of study participants using sterilized cotton swabs. Plaques were collected from the subgingival area using a sterilized curette. Bacterial genomic DNA was extracted using MagNA Pure 96 DNA and Viral NA low-volume kits. Real-time polymerase chain reaction (PCR) was performed using the PowerCheck^TM Periodontitis Pathogens Multiplex Real-time PCR kit. Results: Aggregatibacter actinomycetemcomitans, Prevotella intermedia, and Fusobacterium nucleatum of the orange complex were not observed in the subgingival biofilms of all study participants. For Porphyromonas. gingivalis, a significant correlation was observed between supragingival, subgingival, and total tooth surface biofilms. Compared to the supragingival and subgingival biofilmss, total tooth surface biofilm exhibited the highest bacterial count when the inswabbing method was used. Conclusions: Based on these findings, the supragingival swab method is recommended for oral bacterial research.

• 생명과학회지
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• 제19권7호
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• pp.956-962
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• 2009

구강질환 원인균에 대한 자몽종자추출물(GSE)과 법제유황수(PSS)의 항균활성을 확인하기 위해, 평판배지확산법 및 액체감수성실험법으로 Streptococcus mutans, Prevotella intermedia, Porphyromonas gingivalis 및 Candida albicans에 대한 항균활성을 측정하였다. 평판배지확산법의 경우, GSS의 농도가 40 ${\mu}l$/disk일 때, S. mutans, P. intermedia, P. gingivalis 및 C. albicans에 대한 항균활성은 각각 11.0, 9.5, 8.0, 9.0 mm로 나타났으며, PSS의 농도가 40 ${\mu}l$/disk일 때에는 각각 2.0, 3.5, 0.0, 1.5 mm로 나타났다. 또한 액체배지감수성실험에서 GSE에 대한 MIC 값은 0.24, 0.06, 0.10, 15.63 ${\mu}l$/ml 이었으며, PSS 에 대한 MIC 값은 각각 0.12, 3.91, >125, 7.81 ${\mu}l$/ml이었다. 치약 시제품의 항균효과를 확인하기 위해, GSE와 PSS가 농도별로 함유된 치약시제품 원액과 2배 및 4배 희석액을 제조하여 평판배지확산법을 수행한 결과, 0.5% 치약 시제품의 2배 희석액에서 각각 7.3, 4.3, 2.2 그리고 1.5 mm로 나타났다. 치약 시제품의 물리 화학적 특성을 확인한 결과, pH, 점도, 굴절 및 색도 변화에 있어서 모두 안정성을 보였고, 이상의 결과를 바탕으로 기능성 치약과 같은 구강질환 예방 및 치료제 개발 가능성을 확인하였다.

• Restorative Dentistry and Endodontics
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• 제24권1호
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• pp.1-12
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• 1999

Bacterial infection of the pulp results in the development of a periapical lesion with the concomitant resorption of periapical bone. The cytokines are believed to play an important role in this matter. The purpose of this study was to find the relationship among the presence of black pigmented bacteria, the levels of cytokines(TNF-${\alpha}$, -${\beta}$, IL-$1{\beta}$, and TGF-${\beta}1$), and the amount of bone resorption in periapical and pulpal diseases. For the purpose, the patients were grouped into chronic apical pathosis, acute apical pathosis, acute pulpitis, and a healthy control group. Root canal samples were taken from periapical tissue exudates during routine endodontic treatment, and the venous blood was taken from each patients. The samples were processed to measure local and systemic levels of the cytokines using enzyme linked immunosorbent assay(ELISA). Bacterial content of Porphyromonas endodontalis, Porphyromonas gingivalis, and Prevotella nigrescens were measured by indirect immunofluorescence method and the size of the periapical lesions were measured from the radiographs. The following results were obtained: 1. The levels of bone resorptive cytokines(TNF-${\alpha}$, TNF-${\beta}$, and IL-$1{\beta}$) in exudates from acute and chronic apical pathoses were significantly higher than those from acute pulpitis and the normal pulps(p<0.05). 2. IL-$1{\beta}$ were the highest among the bone resorptive cytokines in apical pathoses. However, no statistical difference between acute and chronic lesions were found(p>0.05). 3. The levels of TGF-${\beta}1$ in exudates from acute pulpitis and chronic apical pathoses were significantly higher than those from acute apical pathoses and the normal pulps(p<0.05). However, there were no significant correlations among the levels of bone resorptive cytokines. 4. The levels of TNF-${\beta}$ in serum were significantly higher than those from the exudates while serum TGF-${\beta}1$ concentrations were significantly lower(p<0.05). 5. Exudates from the canals in which the P. nigrescens were detected showed significantly higher levels of IL-$1{\beta}$ than those from the canals without the microorganism(p<0.05). 6. There were no significant correlations among the levels of the cytokines, the amount of bone destruction, and the presence of acute and chronic symptoms(p>0.05).

• Restorative Dentistry and Endodontics
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• 제24권1호
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• pp.13-25
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• 1999

P. endodontalis which was known to be associated with the infected root canals and periapical lesions is very difficult to detect by culture methods or traditional methods. Detection of bacteria using polymerase chain reaction(PCR) for 16S ribosomal DNA(rDNA) is fast, simple, and accurate with relatively small amount of target cells. 16S rDNA consist of conserved regions those are same to all species, and variable regions which represent species specificity. The 16S rDNA sequences of P. endodontalis and P. gingivalis were aligned and two highly variable regions were selected as a pair of species specific oligonucleotide primers for P. endodontalis. And then the pair of primers for PCR amplification was synthesized to identify P. endodontalis. The sequences of the species specific primers for the 16S rDNA of P. endodontalis were as follows ; sense primer[endo1]: 5'-CTATATTCTTCTTTCTCCGCATGGAGGAGG-3' antisense primer[endo2]: 5'-GCATACCTTCGGTCTCCTCTAGCATAT-3' In this study, for the identification of P. endodontalis without culture from the mixed clinical samples, PCR was done with species specific primers for the 16S rDNA sequences of P. endodontalis. The results were as follows : 1. The species specificity of the primers for the 16S rDNA of P. endodntalis was determined by the PCR methods. About 490bp amplicon which was specific only for P. endodntalis was produced with P. endodontalis. No amplicon was produced by PCR with other strains similar to P. endodontalis. 2. The synthesized species specific primers reacted with conventionally identified P. endodontalis which we have in conservative dentistry laboratory. 3. The identification of P. endodontalis using PCR technique with samples collected from infected root canals or periapical lesions was more sensitive than that of culture methods. 4. Seven samples revealed including P. endodontalis by PCR technique. Five of them were related with pains, two of them with sinus tract, three of them with foul odor, and three of them with purulent drainage. P. endodontalis was shown to have great relation with pains.

• 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.

• 구강회복응용과학지
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• 제38권2호
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• pp.90-96
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• 2022

목적: 이 연구의 목적은 표고 버섯 추출물의 치주 병원체에 대한 항균 활성과 인간 치은 섬유아세포에 대한 세포 독성을 조사하는 것입니다. 연구 재료 및 방법: 표고버섯을 물과 아세톤에 닮근 후, 상등액을 건조하여 추출물을 채취하였다. 추출물에 대한 치주 병원체의 감수성을 조사하였다. 인간 치은 섬유아세포에 추출물을 처리하고 CCK-8 용액을 이용하여 세포 생존율을 측정하였다. 결과: 표고버섯 물추출물은 2.5 mg/ml에서 치주병원체의 증식을 유의하게 감소시켰다(P < 0.05). 아세톤 추출물은 0.32 mg/ml에서 Porphyromonas gingivalis와 Tannerella forsythia의 성장을 유의하게 억제하였고, 0.64 mg/ml에서 Treponema denticola의 성장을 유의하게 억제하였다(P < 0.05). 추출물의 세포독성은 2.5 mg/ml의 농도에서 나타났으며, 1.25 mg/ml 농도의 추출물이 4시간 후부터 세포 생존률을 감소시키는 것으로 나타났다. 결론: 표고버섯 추출물은 치주염 유발 세균에 대한 항균 활성과 염증 완화 효과를 보였다. 따라서 추출물은 치주질환 예방 및 치료에 대한 후보물질이 될 수 있다.