• Journal of Korean society of Dental Hygiene
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• v.24 no.2
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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.

• Journal of Life Science
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• v.19 no.7
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• pp.956-962
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• 2009

The aim of this study was to assess the effects of dentifrice-contatning grapefruit seed extract (GSE) and processed sulfur solution (PSS) on antimicrobial effects against oral pathogens. We first evaluated the antimicrobial effects of GSE and PSS against oral microbes: Streptococcus mutans (Sm), Prevotella intermedia (Pi), Porphyromonas gingivalis (Pg) and Candida albicans (Ca). When antimicrobial activity against Sm, Pi, Pg and Ca was tested, at 40 $\mu$l/disk, the inhibition zones of GSE were 11.0, 9.5, 8.0 and 9.0 mm, respectively. With the same method, the inhibition zones of PSS were 2.0, 3.5, 0.0 and 1.5 mm, respectively. In the micro broth dilution method, the MIC values of GSE against Sm, Pi, Pg and Ca were 0.24, 0.06, 0.10 and 15.63 $\mu$l/rnl, respectively. The MIC values of PSS were 0.12, 3.91,>125 and 7.81 $\mu$l/ml, respectively. When pH, refractive index, viscosity and color value of dentifrice-containing GSE and PSS were measured, there were no significant changes in these physical properties compared to the control samples. Antimicrobial activities of dentifrice products containing 0.5% GSE and 0.5% PSS against oral pathogens were 7.3, 4.3, 2.2 and 1.5 mm, respectively. According to these results, we conclude that there may be a role for GSE and PSS in the development of new oral supplies.

• Restorative Dentistry and Endodontics
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• v.24 no.1
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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 Dental Rehabilitation and Applied Science
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• v.34 no.3
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• pp.186-195
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• 2018

Purpose: We performed quantitative and qualitative analysis of typical periodontal bacteria using real time PCR method to investigate the microbiological difference according to the severity of peri-implant disease in Koreans. Materials and Methods: Total of 60 implants were divided into three groups (healthy group, peri-implant mucositis group, peri-implantitis group) through periapical radiographs and clinical indices. The evaluated clinical parameters were pocket depth, plaque index, suppuration and bleeding on probing. Using a sterilized curette instrument, microbial samples were collected from the subgingival plaque and real-time PCR was performed on five periodontal bacteria. The relative expression levels of microorganisms were compared by comparative delta-CT method. Results: The relative expression levels of E. corrodens and T. denticola were significantly higher in the peri-implantitis group (P < 0.017). On the other hand, the relative expression level of F. nucleatum and P. gingivalis was relatively high in the healthy implant group regardless of the severity of disease. P. intermedia was significantly lower in the healthy implant group (P < 0.017). Conclusion: Periodontal bacteria were detected in Koreans with peri-implant diseases, but there was no microbiological distribution similar to periodontitis.

• Korean journal of food and cookery science
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• v.27 no.3
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• pp.21-28
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• 2011

In this study, we analyzed flower tea activity against oral bacteria. Lagerstroemia indica, Paeonia suffruticosa and Hemerocallis fulva showed high extract yields. Bellis perennis, Punica granatum and Cercis chinensis showed the high rates of yield by ethanol extraction. Extract yield seemed to be related to the characteristics of the specimens rather than to the solvent. Streptococcus mutans, Streptococcus obrinus, Porphyromonas gingivalis and Prevotella intermedia were used to investigate extracts activity against bacteria; the former two cause dental caries and the latter two cause halitosis. Cornus officinalis, L. indica, P. granatum and P.s uffruticosa showed high antibacterial activities against S. mutans. In specimens extracted with ethanol, P. suffruticosa, Camellia sinensis, Camellia japonica L. and Rosa hybrida showed high antibacterial activities. L. indica, P. granatum and C. officinalis showed high antibacterial activities against S. sobrinus. C. officinalis, P. granatum, L. indica and P. suffruticosa showed high activities for specimens extracted with ethanol. The results show that the warm extracts of C. officinalis, L. indica and P. granatum may be effective to prevent dental caries. In particular, the ethanol-based extracts of C. officinalis, P. suffruticosa and C. sinensis were effective to prevent dental caries and thus may be highly marketable. Chrysanthemum zawadskii, R. hybrida, P. granatum, C. japonica L. and Zinnia elegans showed high antibacterial activity against P.gingivalis. R. hybrida showed the highest ethanol extract activity, followed by P. suffruticosa, P. granatum, C. japonica L. and L. indica. R. hybrida, P. granatum, C. morifolium and C. japonica showed high activity against Pr.intermedia in the order named. C. zawadskii, P. granatum, L. indica, C. japonica and A. princeps showed high ethanol extract activity. Thus, the warm extracts of R. hybrida, P. granatum and C. japonica may be helpful to reduce halitosis. In addition, the ethanol-based extracts of P. granatum, C. japonica and L. indica are expected to be highly marketable as mouthwashes.

• Journal of Periodontal and Implant Science
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• v.48 no.4
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• pp.261-271
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• 2018

Purpose: Few studies have examined periodontal pathogens from saliva samples in periodontally healthy young adults. The purposes of this study were to determine the prevalence of periodontopathic bacteria and to quantify periodontal pathogens in saliva samples using real-time polymerase chain reaction (PCR) assays in periodontally healthy Korean young adults under 35 years of age. Methods: Nine major periodontal pathogens were analyzed by real-time PCR in saliva from 94 periodontally healthy young adults. Quantification of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Prevotella intermedia, Fusobacterium nucleatum, Campylobacter rectus, Peptostreptococcus anaerobius, and Eikenella corrodens was performed by DNA copy number measurement. Results: F. nucleatum and E. corrodens were detected in all subjects; the numbers of positive samples were 87 (92.6%), 91 (96.8%), and 90 (95.7%) for P. gingivalis, P. anaerobius, and C. rectus, respectively. Other pathogens were also detected in periodontally healthy subjects. Analysis of DNA copy numbers revealed that the most abundant periodontal pathogen was F. nucleatum, which was significantly more prevalent than all other bacteria (P<0.001), followed by P. anaerobius, P. gingivalis, E. corrodens, C. rectus, and T. denticola. There was no significant difference in the prevalence of each bacterium between men and women. The DNA copy number of total bacteria was significantly higher in men than in women. Conclusions: Major periodontal pathogens were prevalent in the saliva of periodontally healthy Korean young adults. Therefore, we suggest that the development of periodontal disease should not be overlooked in periodontally healthy young people, as it can arise due to periodontal pathogen imbalance and host susceptibility.

• Journal of Periodontal and Implant Science
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• v.27 no.4
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• pp.723-737
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• 1997

본 연구의 목적은 조직유도재생술의 초기치유시에 구강양치액으로 사용되어지는 0.1% 클로르헥시딘과 0.2% 클로르헥시딘을 사용했을 경우, 양치액을 사용하지 않았을 경우의 세균감염 정도를 비교하는 것이다. 30명의 성인형 치주염에 이환되어진 사람을 대상으로 하였다. 초기치료(Scaling/Root planing/Oral hygiene instruction)를 시행한 후에 한 사람에 한 군데씩 선정하여 2급이나 3급의 치근이개부를 가지고 임상적으로 혹은 방사선학적으로 치간골내낭을 보이지 않는 치아에 통법에 따라 Gore-TexTM를 위치시켰다. 술후 5일간 항생제 (UnasynTM 375mg tablet p.o.tid)를 투여하고 차폐막을 제거할 때까지(4주 혹은 6주) 10명의 환자에게는 0.1% 클로르헥시딘을, 다른 10명의 환자에게는 0.2% 클로르헥시딘으로 구강양치를 하게 하고, 또 다른 10명의 환자에게는 구강양치액을 사용하지 않도록 하였다. 또 1주일에 한번씩 전문가구강위생술식을 실시하였다. 4주나 6주 후에 차폐막을 제거하고 주사전자현미경, 혐기성 세균배양을 이용하여 세균감염정도를 비교하였다. 1. 주사전자현미경으로 관찰시에 0.1% 클로르헥시딘을 사용했을 경우와 0.2% 클로르헥시딘을 사용했을 경우, 클로르헥시딘을 사용하지 않은 경우에 별 차이를 발견할 수 없었다. 2. 혐기성 세균배양시에 0.2% 클로르헥시딘을 사용했을 경우, 0.1%클로르헥시딘을 사용했을 경우보다 적은 수의 세균 수를 보였으나 통계적으로 유의할 만한 차이는 보이지 않았다. 클로르헥시딘을 사용하지 않은 경우에는 다른 두 경우에 비해 통계적으로 유의할 만한 차이를 보였다.(P<0.05) 3. Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia를 인지한 경우에는 세 경우 모두 비슷한 비율로 발견되었다.

• Journal of Periodontal and Implant Science
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• v.44 no.6
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• pp.280-287
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• 2014

Purpose: The purpose of this randomized single-blind controlled trial was to elucidate the clinical and antimicrobial effects of daily phototherapy (PT) as an adjunct to scaling and root planing (SRP) in patients with chronic periodontitis. Methods: The study was conducted from December 2013 to May 2014 at Ewha Womans University Mokdong Hospital, Seoul, Korea. Forty-one patients with mild to moderate chronic periodontitis were randomly divided into two therapeutic groups in a 1:1 ratio: SRP+PT and SRP (control) groups. All participants underwent full-mouth SRP. PT was performed thrice a day for a month by using electric toothbrushes with embedded light-emitting diodes. Plaque index, gingival index, probing pocket depth (PPD), clinical attachment level (CAL), and bleeding on probing were assessed before (baseline) and four weeks after (follow-up) the treatment. Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Prevotella intermedia, Fusobacterium nucleatum, Parvimonas micra, Campylobacter rectus, Eikenella corrodens, Streptococcus mutans, and Streptococcus sobrinus levels were detected by a real-time polymerase chain reaction at the same points in time. Results: The clinical parameters improved in both the groups. At the follow-up assessment, PPD was significantly decreased in the SRP+PT group (P=0.00). Further, PPD and CAL showed significantly greater changes in the SRP+PT group than in the SRP group (PPD, P=0.03; CAL, P=0.04). P. gingivalis and T. forsythia levels decreased in this group, but no significant intergroup differences were noted. Conclusions: Adjunctive PT seems to have clinical benefits, but evidence of its antimicrobial effects is not sufficient. Long-term studies are necessary to develop the most effective PT protocol and compare the effectiveness of PT with and without exogenous photosensitizers.

• The Journal of Advanced Prosthodontics
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• v.12 no.4
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• pp.233-238
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• 2020

PURPOSE. This study aims to compare the marginal fitness of two types of implant-supported fixed dental prosthesis, i.e., cementless fixation (CL.F) system and cement-retained type. MATERIALS AND METHODS. In each group, ten specimens were assessed. Each specimen comprised implant lab analog, titanium abutment fabricated with a 2-degree tapered axial wall, and zirconia crown. The crown of the CL.F system was retained by frictional force between abutment and relined composite resin. In the cement-retained type, zinc oxide eugenol cement was used to set crown and abutment. All specimens were sterilized with ethylene oxide, immersed in Prevotella intermedia culture in a 50 mL tube, and incubated with rotation. After 48 h, the specimens were washed thoroughly before separating the crown and abutment. The bacteria that penetrated into the crown-abutment interface were collected by washing with 500 µL of sterile saline. The bacterial cell number was quantified using the agar plate count technique. The BacTiter-Glo Microbial Cell Viability Assay Kit was used to measure bacterial adenosine triphosphate (ATP)-bioluminescence, which reflects the bacterial viability. The t-test was performed, and the significance level was set at 5%. RESULTS. The number of penetrating bacterial cells assessed by colony-forming units was approximately 33% lower in the CL.F system than in the cement-retained type (P<.05). ATP-bioluminescence was approximately 41% lower in the CL.F system than in the cement-retained type (P<.05). CONCLUSION. The CL.F system is more resistant to bacterial penetration into the abutment-crown interface than the cement-retained type, thereby indicating a precise marginal fit.

• Journal of Periodontal and Implant Science
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• v.32 no.1
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• pp.249-255
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• 2002

Zea Mays L. has been known to be effective for improving tissue health and Magnoliae cortex to have effective antibacterial and antimicrobial activity against pathogenic microbes. The purpose of this study was to examine the antimicrobial effects of Zea Mays L. and Magnoliae cortex extract mixtures on periodontal pathogens(Prevotella intermedia, Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Streptococcus mutans )and to examine the effects on human gingival fibroblast cellular activity. Zea Mays L. and Magnoliae cortex extracts and their mixtures were prepared with various mixing ratios (0.5:1, 1:1, 1.5:1, 2:1). These extracts were loaded to periodontal pathogen cultured petri dish for antimicrobial test and also loaded to cultured human gingival fibroblast for cellular activity test. Each test was repeated 3 times and data were analyzed by one-way ANOVA with 95% confidence level. Mixture of these two extracts showed greater amount of inhibition area on periodontal pathogen and more improved gingival fibroblast activity as Zea Mays L. ratio reduced. So, mixture ratio 0.5:1 (Zea Mays L. : Magnoliae cortex) group showed statistical significance in antimicrobial activity and cellular activity among various mixtures(p < 0.05). In conclusion, 0.5:1 (Zea Mays L. : Magnoliae cortex) mixture possessed best gingival fibroblast cellular activity and antimicrobial activity toward periodontal pathogens.