• Journal of Microbiology and Biotechnology
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• v.34 no.2
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• pp.289-295
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• 2024

We have developed an aptamer that specifically binds to Porphyromonas gingivalis to reduce the cellular damage caused by P. gingivalis infection and applied it as a biosensor. P. gingivalis is one of the major pathogens causing destructive periodontal disease among the periodontal microorganisms constituting complex biofilms. Porphyromonas gingivalis G-protein (PGP) known to play an important role in the transmission of germs was used as a target protein for the screening of aptamer. The aptamer that has binds to the G-protein of P. gingivalis, was screened and developed through the Systemic Evolution of Ligands by Exponential Energy (SELEX) method. Modified-Western blot analysis was performed with the aptamer which consisted of 38 single-stranded DNA to confirm the selectivity. ELONA (enzyme linked oligonucleotide assay) used to confirm that the aptamer was sensitive to PGP even at low concentration of 1 ㎍/ml. For the rapid detection of P. gingivalis, we constructed a surface plasmon resonance biosensor with SPREETA using the PGP aptamer. It was confirmed that PGP could be detected as low concentration as at 0.1 pM, which is the minimum concentration of aptamer sensor within 5 min. Based on these results, we have constructed a SPREETA biosensor based on aptamer that can bind to P. gingivalis G-protein. It can be used as an infection diagnosis system to rapidly diagnose and analyze oral diseases caused by P. gingivalis.

• Biomedical Science Letters
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• v.29 no.2
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• pp.88-94
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• 2023

In oriental medicine, the fruit of Actinidia polygama has long been used to alleviate the symptoms of gout, arthritis, and inflammation. In this study, it was to designed to analyze the antibacterial activity of A. polygama ethanol extract (APEE) against Streptococcus mutans, one of the major strains for dental caries, and Porphyromonas gingivalis, one of the critical strains for periodontal disease. The antibacterial activity of APEE was analyzed by disk diffusion, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) assays. In addition, it was also analyzed the inhibitory effect of APEE on bacterial growth and biofilm formation against both oral pathogens. APEE exhibited its antibacterial effect through the inhibited bacterial diffusion as well as low concentration of MIC and MBC. In addition, APEE significantly inhibited not only bacterial growth but also biofilm formation in a dose-dependent manner. Consequently, APEE showed potent antibacterial activity against both S. mutans and P. gingivalis, which indicates that APEE might be used as a potential antibacterial material for the improvement of oral healthcare.

• Journal of Periodontal and Implant Science
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• v.32 no.2
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• pp.403-414
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• 2002

I. Purpose of Study Zea Mays L. has been known to be effective for improving periodontal health and Magnoliae cortex to have effective antibacterial and antimicrobial activity against periodontal pathogens. The purpose of this study was to examine the biologic effects of Zea Mays L. and Magnoliae cortex extract mixtures on healing of rat calvarial bone defects. II. Materials & Methods 8mm circular defects were prepared on rat calvaria during surgical procedures of 180 Sprague-Dawley rats. The ethanolic extracts of Magnoliae cortex and Zea Mays L. and these two natural extract 1:1 and 2:1 (Magnoliae: Zea Mays L.) ratio mixtures were oral administrated by oral zondes once a day at two different dose of 94.5mg/kg, 189mg/kg body weight. There are nine groups of rats in this study: control group (no sample loading), Magnoliae cortex extract loading groups (I,II)(94.5mg/kg,189mg/kg respectively), Zea Mays L. extract loading groups (I,II), M:Z(1:1) loading groups (I,II), M:Z(2:1) loading groups(I,II). Rats were sacrificed at 4 weeks and 6 weeks after surgery. New bone formations around calvarial defects were radiographically and histologically measured by computerassisted histomorphometry. Each data was statistically analyzed by One-way ANOVA test. III. Results There were statistical significances between negative control group and the other test groups on radiographical and histological quantitative assessments. Among test groups, mixture groups showed statistical significances, especially, M:Z (2:1) groups (I and II) were highly significant.(p<0.05) These results implicated that the mixture of Magnoliae and Zea Mays L. (2:1 mixing ratio) with 94.5mg/kg concentration might be highly effective on the wound healing of bony defected site and have potential possibilities as a useful drug to promote bone tissue regeneration.

• The Korea Journal of Herbology
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• v.23 no.2
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• pp.1-8
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• 2008

Objectives : The purpose of this study was to evaluate on the antimicrobial effect on the periodontal pathogens and anti-inflammatory effect of Artemisiae Iwayomogii Herba. Artemisiae Iwayomogii Herba has been used for treating as Artemisiae Capilaris Herba in Korea. Methods : Artemisiae Iwayomogii Herba was prepared by extracting medicinal herb with water. We investigated antimicrobial activity by the minimun inhibitory concentration (MIC) test. We also investigated inhibition of IL-$1{\beta}$-induced collagenase-l(MMP-l), stromelysin-1(MMP-3), interleukin-6 gene expression in human gingival fibroblasts. Results : The antimicrobial effect of Artemisiae Iwayomogii Herba was evaluated with MIC against periodontopathogens; Porphyromonas gingivalis 2561, W50, A7A1-28, 9-14K-1, Prevotella intermedia28, and Actinobacillus actinomycetemcomitans Y4, MICs of Artemisiae Iwayomogii Herba were 0.156 mg/ml, 0.625 mg/ml, 0.313 mg/ml, 1.25 mg/ml, 10 mg/ml and 10 mg/ml. The anti-inflammatory effect of Artemisiae Iwayomogii Herba was evaluated with Influence of herbs on the IL-$1{\beta}$-induced expression of MMP-1, MMP-3, interleukin-6, IL-$1{\beta}$ increased MMP-1, MMP-3, interleukin-6 mRNA levels. Artemisiae Iwayomogii Herba significantly inhibited IL-$1{\beta}$-induced MMP-1, MMP-3, interleukin-6 gene expressions in a dose-dependent manner. Conclusions : These results suggested that Artemisiae Iwayomogii Herba might reduce the excessive proteolytic capacity of the gingival fibroblast during inflammation and could be developed a new drug in periodontitis.

• The Journal of Korean Medicine
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• v.29 no.2
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• pp.182-192
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• 2008

Objective: The purpose of this study was to evaluate on the antimicrobial effect on the periodontal pathogens and anti-inflammatory effect of Eriobotryae folium. Eriobotryae folium are constituent herbs of Gagamgamroum, which has been used for a long time in oriental medicine as a herbal medicine for treating halitosis and toothache. Method: Eriobotryae folium was prepared by extracting medicinal herb with water. We investigated antimicrobial activity by the minimum inhibitory concentration (MIC) test. We also investigated inhibition of $IL-1{\beta}-induced$ collagenase (mmp-1), stromelysin-1 (mmp-3), interleukin-6 gene expression in human gingival fibroblasts using RTPCR analysis. Result: The antimicrobial effects of Eriobotryae folium was evaluated with MIC against periodontopathogens; Porphyromonas gingivalis 2561, W50, A7A1-28, 9-14K-1, Prevotella intermedia 28, and Actinobacillus actinomycetemcomitans Y4. MICs of Eriobotryae folium were 1.25 mg/ml, 2.5 mg/ml, 0.625 mg/ml, 1.25 mg/ml, 10 mg/ml and 10 mg/ml. The anti-inflammatory effect of Eriobotryae folium was evaluated with influence of herbs on the $IL-1{\beta}-induced$ expression of mmp-1, mmp-3, and interleukin-6. $IL-1{\beta}$ increased mmp-1, mmp-3, and interleukin-6 mRNA levels. Eriobotryae folium significantly inhibited $IL-1{\beta}-induced$ mmp-1, mmp-3, and interleukin-6 gene expressions in a dose-dependent manner. Conclusion: These results suggested that Eriobotryae folium might reduce the excessive proteolytic capacity of the gingival fibroblast during inflammation and could be developed as a new drug for periodontitis.

• The Korean Journal of Physiology and Pharmacology
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• v.19 no.1
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• pp.51-57
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• 2015

The etiology of periodontal disease is multifactorial. Exogenous stimuli such as bacterial pathogens can interact with toll-like receptors to activate intracellular calcium signaling in gingival epithelium and other tissues. The triggering of calcium signaling induces the secretion of pro-inflammatory cytokines such as interleukin-8 as part of the inflammatory response; however, the exact mechanism of calcium signaling induced by bacterial toxins when gingival epithelial cells are exposed to pathogens is unclear. Here, we investigate calcium signaling induced by bacteria and expression of inflammatory cytokines in human gingival epithelial cells. We found that peptidoglycan, a constituent of grampositive bacteria and an agonist of toll-like receptor 2, increases intracellular calcium in a concentration-dependent manner. Peptidoglycan-induced calcium signaling was abolished by treatment with blockers of phospholipase C (U73122), inositol 1,4,5-trisphosphate receptors, indicating the release of calcium from intracellular calcium stores. Peptidoglycan-mediated interleukin-8 expression was blocked by U73122 and 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis (acetoxymethyl ester). Moreover, interleukin-8 expression was induced by thapsigargin, a selective inhibitor of the sarco/endoplasmic reticulum calcium ATPase, when thapsigargin was treated alone or co-treated with peptidoglycan. These results suggest that the gram-positive bacterial toxin peptidoglycan induces calcium signaling via the phospholipase C/inositol 1,4,5-trisphosphate pathway, and that increased interleukin-8 expression is mediated by intracellular calcium levels in human gingival epithelial cells.

• International Journal of Oral Biology
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• v.39 no.1
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• pp.15-22
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• 2014

Aggregatibacter actinomycetemcomitans is an important pathogen in the development of localized aggressive periodontitis. Lipopolysaccharide (LPS) is a virulent factor of periodontal pathogens that contributes to alveolar bone loss and connective tissue degradation in periodontal disease. Our present study was designed to investigate the cytokine expression and signaling pathways regulated by A. actinomycetemcomitans LPS (Aa LPS). Cytokine gene expression profiling in RAW 264.7 cells was performed by microarray analyses. The cytokine mRNA and protein levels and related signaling pathways induced by Aa LPS were measured by RT-PCR, ELISA and western blotting. Microarray results showed that Aa LPS strongly induced the expression of NF-${\kappa}B$, NF-${\kappa}B$-related genes, inflammatory cytokines, TNF-${\alpha}$ and IL-$1{\beta}$ in RAW 264.7 cells. NF-${\kappa}B$ inhibitor pretreatment significantly reduced the levels of TNF-${\alpha}$ and IL-$1{\beta}$ mRNA and protein. In addition, the Aa LPS-induced TNF-${\alpha}$ and IL-$1{\beta}$ expression was inhibited by p38/JNK MAP kinase inhibitor pretreatment. These results show that Aa LPS stimulates TNF-${\alpha}$ and IL-$1{\beta}$ expression through NF-${\kappa}B$ and p38/JNK activation in RAW 264.7 cells, suggesting the essential role of this pathway in the pathogenesis of localized aggressive periodontitis.

• Journal of Korean society of Dental Hygiene
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• v.12 no.3
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• pp.631-640
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• 2012

Objectives : This study mean to confirm the antibacterial activity of a garlic extract widely culturing in our region and was to determine the effect of dentifrice containing 0.1% extracts of garlic on dental plaque and gingivitis in a double blind and clinical studies in 50 healthy adults aged from 20 to 22 years who provided a consent for their participation. Methods : The antibacterial activity was evaluated using triple distilled water and the Kirby-Bauer disk diffusion method and the minimal inhibitory concentration(MIC) against various pathogens for periodontal disease, such as P. gingivalis 381(ATCC33277), was estimated. The experimental groups classified according to the concentration of garlic extract used: 10,000ppm(A), 5,000ppm(B), 2,500ppm(C), 1,000ppm(D). Oral examination of subjects was performed through clinical periods and on day of baseline, 6, 12, 19, 25 days plaque index and gingival index were scored by Turesky' modified index and Loe & Silness index. After 12, 19, 25 days use of their respective dentifrices, statistically decreases of plaque index, gingival index were shown in both the experimental and the control group, respectively. Results : There was significant antibacterial activity in the "2,500ppm(C)" group against P. gingivalis 381. Experimental group exhibited significantly the lower plaque levels and the higher levels of gingival health by the use of the dentifrices contained extract of garlic from 6 days compare with control group(p<0.05). The degree of decrease was more significant on gingivitis level of the experimental group than the control group(p<0.05). Conclusions : This findings indicated that the oral products containing a garlic extract is effective in preventing and treating periodontal diseases, and has potential value in inhibiting periopathogens.

• Journal of Korean society of Dental Hygiene
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• v.17 no.1
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• pp.155-168
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• 2017

Objectives: The purpose of this research is to examine oral pathogen distribution among elderly with dementia in a care facility to understand the importance of preventively managing oral diseases in terms of preventively managing senile diseases. Methods: From 11th August 2015 to 11th October 2015, gingival crevicular fluid was collected from 130 subjects consisting of demented/non-demented elderly people aged above 65 in care facilities located in the regions of Busan/Gyeonggnam. Based on collected data, real-time PCR analysis on oral pathogen was conducted. Results: The demented elderly group consisting of female patients aged from 0 to 79 indicated higher ratios of T. denticola in comparison to the non-demented elderly group, and the demented elderly group consisting of female patients aged above 80 indicated a high ratio of S. mutans. It was confirmed that P. gingivalis and T. forsythensis categorized under the red complex are correlated, and that bacterial species categorized under the orange complex and bacterial species categorized under the red complex are correlated. Conclusions: Because eldery people with demntia are exposed to periodontal disease and dental caries, their oral environments are more vulnerable. In order to improve such environments, it is necessary to provide care facility personnel with an opportunity to receive education to become aware of the importance of oral health, and it is also necessary to compulsorily assign dental hygienists to care facilities so that they can serve as personnel specializing in oral health management.

• Journal of Periodontal and Implant Science
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• v.42 no.6
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• pp.217-223
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• 2012

Purpose: Several parameters have been described for determining the success or failure of dental implants. The surface properties of transgingival implant components have had a great impact on the long-term success of dental implants. The purpose of this study was to compare the tendency of two periodontal pathogens to adhere to and colonize zirconia abutments and titanium alloys both in hard surfaces and soft tissues. Methods: Twelve patients participated in this study. Three months after implant placement, the abutments were connected. Five weeks following the abutment connections, the abutments were removed, probing depth measurements were recorded, and gingival biopsies were performed. The abutments and gingival biopsies taken from the buccal gingiva were analyzed using real-time polymerase chain reaction to compare the DNA copy numbers of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, and total bacteria. The surface free energy of the abutments was calculated using the sessile water drop method before replacement. Data analyses used the Mann Whitney U-test, and P-values below 0.05 find statistical significance. Results: The present study showed no statistically significant differences between the DNA copy numbers of A. actinomycetemcomitans, P. gingivalis, and total bacteria for both the titanium and zirconia abutments and the biopsies taken from their buccal gingiva. The differences between the free surface energy of the abutments had no influence on the microbiological findings. Conclusions: Zirconia surfaces have comparable properties to titanium alloy surfaces and may be suitable and safe materials for the long-term success of dental implants.