• Journal of Periodontal and Implant Science
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• v.29 no.1
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• pp.265-276
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• 1999

This study was performed to evaluate the effect of hydrogen peroxide-producing Lactobacillus acidophilus V-20onthe replication of periodontal pathogens, Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis. When A. actinomycetemcomitans and P. gingivalis were incubated alone and in the combination with L. acidophilus V-20, the viable cell numbers of A. actinomycetemcomitans and P. gingivalis were compared between those cultures. The effect of S. mutans, E. durans, and L. lactis on the replication of A. actinomycetemcomitans and P. gingivalis was also evaluated. The change of periodontal indexes(probine depth, gingival index, GCF volume) and the viable cell numbers of A. actinomycetemcomitans and black pigmented bacteroides in subgingival plaque sample were evaluated following gargling of fermented milk made from L. acidophilus V-20 for 1 month on patients with periodontal disease in maintenance phase. In the mixed culture of L. acidophilus V-20 and A. actinomycetemcomitans or P. gingivalis, the replication of A. actinomycetemcomitans or P. gingivalis wascompletely inhibited. But in the mixed culture of P. gingivalis and hydrogen peroxide-nonproducing Lactobacillus casei, the viable ceil numbers of P. gingivaliswas not decreased when compared with the numbers in the mixed culture of P. gingivalis and L. acidophilus V-20. In the mixed culture of A. actinomycetemcomitans and S. mutans, E. durans, or L. lactis, the viable cell number of A. actinomycetemcomitans was not almost changed when compared with the numbers in the culture of A. actinomycetemcomitans alone. And in the mixed culture of P. gingivalis and E. durans or L. lactis, the viable cell numbers of P. gingivaliswas not almost changed compared with the counts in the culture of P. gingivalis alone. But the replication of P. gingivalis was completely inhibited in the mixed culture of P. gingivalis and S. mutans. When the change of periodontal indexes following gargling of fermented milk was compared with baseline, probing depth and gingival index were not changed, but GCF volume was significantly decreased(p<0.05). And when the viable ceil numbers of microorganisms in subgingival plaque sample were compared with baseline, total viable ceil number was almost unchanged and the viable cell numbers of A. actinomycetemcomitans and black pigmented bacteroides were significantly decreased(p<0.05). These results suggest that L. acidophilus V-20 inhibit the replication of A. actinomycetemcomitans and black pigmented bacteroides by the formation of hydrogen peroxide.

• Journal of Periodontal and Implant Science
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• v.26 no.2
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• pp.542-556
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• 1996

Previous studies have shown that Magnoliae cortex and Ginkgo biloba extracts were showed on the antimicrobial and anti-inflammatory action, in vitro. The purpose of this study was to evaluate on the effect of antimicrobial and anti-inflammatory activity of Magnoliae cortex and Ginkgo biloba extracts containing dentifrice in gingivitis. 70 subjects with gingivitis were divided into an experimental group which performed normal oral hygiene procedure with Magnoliae cortex and Ginkgo biloba extracts containing dentifrice and a control group which also performed normal oral hygiene procedure with the same dentifrice without the natural extracts and completed a doubleblind, cross-over study. At baseline and 3 weeks, subjects were assayed for clinical study by plaque index, gingival index, pocket depth, GCF rate, and microbiological study by subgingival dental plaque bacterial morphotypes by phase contrast microscopy, total anaerobes, total aerobes, Black pigmented bacteroides, A.actionomycetemcomitans, A.viscosus, C.rectus, Ssenguis; P.gingivalis, P.intennedia by bacterial culture and immunofluorescence microscopy. After 3 weeks using their respective dentifrices, reductions in the clinical indices of subjects were similar between the experimental dentifrice group and a control dentifrice group except for statistically significant much reductions in PI, GI, and GCF rate in the experimental dentifrice group as compared to control dentifrice group. Also statistically significant reductions in the motile rods and Spirochetes were found in both experimental group to compare with control group, however statistically much reduction in total anaerobes, Black pigmented bacteroides, and P.gingivalis, P.intennedia were found in the experimental dentifrice group as compared to control dentifrice group. This results indicates that Magnoliae cortex and Ginkgo biloba extracts containing dentifrice might be useful for elimination of gingival inflammation.

• Restorative Dentistry and Endodontics
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• v.23 no.1
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• pp.328-338
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• 1998

Porphyromonas endodontalis, an anaerobic Gram negative cocobacillus which was known to be associated with the infected root canals and periapical lesions, is very difficult to culture and to detect by the traditional method in that it requires much time to induce the specific black pigmentation, and it is very sensitive to oxygen and the antibiotics added in the culture medium. In this study, the nucleotide sequences of the 'probe h' (0.73kb), one of the specific DNA probes top. endodontalis (ATCC 35406) which had been developed by our department, was determined and then a pair of primers for PCR amplification was fabricated to identify P. endodontalis. The plasmids containing 'probe h' were purified by $Wizard^{TM}$ Midipreps DNA Purification System (Promega Corp.), and the nucleotide sequences of the 'probe h' were determined by the dideoxy chain termination method using TaqTrack Sequencing System (Promega Corp.) and detected by fluorescent labelling method. The sense/antisense PCR primers were designed with computer software (Lasergene, DNASTAR Ind. PCR was done with a programmable GeneAmp PCR System 2400 (Perkin Elmer-Cetus Co.). Each sample containing the whole genomic DNA of P. endodontalis and other black-pigmented Bacteroides was itailly denatured at $94^{\circ}C$ for 5 min and then subjected to 30 cycles, each of them consisting of 60s at $94^{\circ}C$, 60s at $60^{\circ}C$, and 90s. at $72^{\circ}C$. The amplified DNA was resolved electrophoretically in a 1.0 % agarose gel in 1X TAE buffer, stained with EtBr, and photographed on a UV transilluminator. The results were as follows : 1. The nucleotide sequences of 'probe h' (743 base pairs) were obtained by dideoxy chain termination method, and from that results the specific primers to P. endodontalis (ATCC 35406), 'Primer H1/ Primer H2', were designed. 2. It has been found that 'Primer H1/H2' could detect P. endodontalis (ATCC 35406) using PCR. 3. The PCR system with this primers may be a powerful technique to amplify the specific sequences of 'probe h' of P. endodontalis (ATCC 35406) that produce distinct identification of it from other black-pigmented Bacteroides, and this could help us to determine the nature of periapical disease.

• Journal of Periodontal and Implant Science
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• v.29 no.4
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• pp.737-749
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• 1999

Recently many researches on plaque removal effect and therapeutic effect of toothpaste containing natural medicines are being studied in early periodontal diseases. The purpose of this study is to examine the clinical and microbiological effect of toothpaste containing natural medicines such as camomile, rhatany, myrrh，sage oil, glycyrrhetinic acid and vitamin E. Sixty three subjects with gingivitis were divided into an experimental group which performed normal oral hygiene procedure with toothpaste containing natural medicines and vitamine E and a control group which also performed normal oral hygiene procedure with Syrinmed? toothpaste without containing herbal extracts and vitamine E. At the baseline, 2 weeks, and 4 weeks, subjects were analyzed for clinical study and microbiological study. After 2 weeks and 4 weeks use of their respective toothpastes, statistically significant decreases of gingival index, plaque index, and bleeding index were shown in both the control and the experimental group. The degree of decrease was more significant in the experimental group than the control group. A statistically significant decrease of pocket depth, and gingival crevicular fluid were shown in both the control and lie experimental group. A statistically significant increase of cocci was shown in both the control and the experimental group, the degree of increase was more significant in the experimental group than control group. A statistically significant decrease ofnon-motile rods, and motile rods were shown in both the control and the experimental group, the degree of decrease was more significant in the experimental group than the control group. Spirochetes increased weakly in both the control and the experimental group but a statistic significance was not shown. A statistically significant decrease of anaerobic bacteria, aerobic bacteria, and black pigmented Bacteroides were shown in both the control and the experimental group. These results indicate that the use of toothpaste containing natural medicines is effective in the prevention and the treatment of periodontal diseases.