• Journal of Periodontal and Implant Science
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• v.43 no.4
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• pp.183-190
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• 2013

Purpose: At present, information regarding periodontal disease in geriatric patients is scarce. The purpose of this study was to quantify the periodontal pathogens present in the saliva of Korean geriatric patients and assess the relationship between the bacterial levels and the periodontal condition. Methods: Six putative periodontal pathogens were quantified by using a real-time polymerase chain reaction assay in geriatric patient groups (>60 years) with mild chronic periodontitis (MCP), moderate chronic periodontitis (MoCP), and severe chronic periodontitis (SCP). The copy numbers of Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, and Prevotella intermedia were measured. Results: It was found that the bacterial copy numbers increased as the severity of the disease increased from MCP to SCP, except for P. intermedia. For P. intermedia, it was found that samples in the MCP group yielded the largest amount. It was also found that the quantities of P. gingivalis, T. forsythia, and T. denticola, the so-called "red complex" bacteria, were lower than those of F. nucleatum, A. actinomycetemcomitans, and P. intermedia in all of the samples. Conclusions: Collectively, the results of this study suggest that the levels of P. gingivalis, T. forsythia, F. nucleatum, and T. denticola present in saliva are associated with the severity of periodontal disease in geriatric patients.

• Journal of Periodontal and Implant Science
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• v.30 no.3
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• pp.661-676
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• 2000

Gingivitis and periodontitis are infectious diseases in that microorganisms are the primary extrinsic cause of the diseases. the occurrence of gingivitis has been associated clearly with the presence of microorganisms at the disease site, and the histologic nature of the tissue involved is indicative of an inflammatory response induced by microorganisms. additional evidence for the microbial etiology of periodontal disease is that numerous antimicrobial agents are effective in reducing plaque accumulation and periodontal diseases. the purpose of this article is to analyze the antimicrobial effects of Pulsatilla koreana. Well-dried Pulsatilla koreana purchased from herbs distributor was ground and extracted into methanol(MeOH), ethylacetate(EtoAc), chlorform($CHCl_3$) and Butyl alcohol(BuOH). we have then applied each solution to the bacteria samples(Bacteroides forsythus, Streptococcus mutans, Streptococcus sanguis, Porphylomonas gingivalis, Actinobacillus actinomycetemcomitans, Eikenella corrodens, Prevotella intermedia, Actinomyces viscosus, Prevotella nigrescens , Rothia dentocariosa, Fusobacterium nucleatum, Pseudomonas aeruginosa, Staphylococcus aureus) collected from several organizations. To conduct susceptibility test(Kirby-Bauer method), plate contained each periodontopathic bacteria is spread extracted into methanol(MeOH), ethylacetate(EtoAc), chlorform($CHCl_3$) and Butyl alcohol(BuOH) and to measure the minimum inhibition concentration(MIC) of the bacteria against the solutions to ultimately determine antimicrobial effects of the solutions, insert bacteria sample into $20{\mu\ell}/{m\ell}$, $10{\mu\ell}/{m\ell}$, $5{\mu\ell}/{m\ell}$, $2.5{\mu\ell}/{m\ell}$ of each solution and control group(not contained solution) 1. Solution extracted into methanol did not show clear zone against all bacteria samples. Only P.nigrescens, S. mutans and S. sanguis in solution extracted into ethylacetate, S. mutans and S. anguis in solutions extracted into chlorform and Butyl alcohol showed clear zone against all bacteria samples. Solution extracted into Butyl alcohol showed clear zone against 13 types of bacteria, excluding P. gingivalis. 2. In Solution extracted into methanol, the bacteria samples grew in the highest concentrated plate, showing minimal variation from control group. 3. In Solution extracted into Butyl alcohol, S. aureus, P. intermedia, E. corrodens, A. actinomycetemcomitans, B. forsythus, P. gingivalis et al. showed decreased growth in the highest concentrated plate. P. auruginosa, R. dentocariosa, A. viscosus, P. nigrescens, S. mutans et al. showed decreased growth at MIC $20{\mu\ell}/{m\ell}$ and S. sanguis showed decreased growth at MIC $10{\mu\ell}/{m\ell}$. 4. By analyzing the MIC level through considering the results from Kirby-Bauer method, Solution extracted into methanol did not reveal any antimicrobial effects and Solution extracted into Butyl alcohol showed the highest antimicrobial effects In conclusion, it can be used the extracts of Pulsatilla koreana as wide spectrum antimicrobial agent.

• Journal of Korean Academy of Oral Health
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• v.41 no.4
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• pp.290-295
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• 2017

Objectives: Dental plaque emits red fluorescence under a visible blue light near the ultra-violet end of the light spectrum. The fluorescence characteristics of each microorganism have been reported in several studies. The aim of this study was to evaluate changes in red fluorescence of oral microorganisms that is affected by blood in the culture media. Methods: The gram-positive Actinomyces naeslundii (AN, KCTC 5525) and Lactobacillus casei (LC, KCTC 3109) and gram negative Prevotella intermedia (PI, KCTC 3692) that are known to emit red fluorescence were used in this study. Each bacterium was activated in broth and cultivated in different agar media at $37^{\circ}C$ for 7 days. Tryptic soy agar with hemin and vitamin $K_3$ (TSA), TSA with sheep blood (TSAB), basal medium mucin (BMM) medium, and BMM with sheep blood (BMMB) were used in this study. Fluorescence due to bacterial growth was observed under 405-nm wavelength blue light using the quantitative light-induced fluorescence-digital (QLF-D) device. The red, green, and blue fluorescence values of colonies were obtained using image-analysis software and the red to green ratio (R/G value) and red to total RGB ratio (R/RGB value) were calculated for quantitative comparison. Results: The QLF-D images of the AN, LC, and PI colonies showed red fluorescence in all media, but the fluorescence of all bacteria was reduced in TSA and BMM media, compared with in TSAB and BMMB media. Both the R/G and the R/RGB values of all bacteria were significantly reduced in growth media without blood (P<0.001). Conclusions: Based on this in vitro study, it can be concluded that red fluorescence of oral bacteria can be affected by growth components, especially blood. Blood-containing medium could be a significant factor influencing red fluorescence of oral bacteria. It can be further hypothesized that bleeding in the oral cavity can increase the red fluorescence of dental plaque.

• International Journal of Oral Biology
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• v.38 no.4
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• pp.141-147
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• 2013

It has been established that berberine has strong antimicrobial effects. Little is known however regarding the antimicrobial activity of berberine against endodontic pathogenic bacteria or its cytotoxicity in human oral tissue cells. The antibacterial properties of berberine were tested against 5 strains of Enterococcus faecalis and type strains of Aggregatibacter actinomycetemcomitans, Prevotella nigrescens, Prevotella intermedia, and Tannerella forsythia, which are involved in endodontic infections. Antimicrobial activity was evaluated through minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) measurements. The viability of normal human gingival fibroblast (NHGF) cells after exposure to berberine was measured using a methyl thiazolyl tetrazolium (MTT) assay. The data showed that berberine has antimicrobial effects against A. actinomycetemcomitans with an MIC and MBC of $12.5{\mu}g/ml$ and $25{\mu}g/ml$, respectively. In the cytotoxicity studies, cell viability was maintained at 66.1% following exposure to $31.3{\mu}g/ml$ berberine. Overall, these findings suggest that berberine has antimicrobial activity against the tested bacteria. Nevertheless, lower concentrations in combination with other reagents will need to be tested before these in vitro results can be translated to clinical use.

• Journal of Microbiology
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• v.43 no.3
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• pp.260-265
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• 2005

The objective of this study was to detect and compare the presence of periodontopathogens in the subgingival plaques of gingivitis lesions in adults who wore fixed orthodontic appliances, as opposed to adults who did not wear any orthodontic appliances. Thirty-six individuals participated in this study. Ninteen of these subjects did not wear any orthodontic appliances, and these subjects comprised the control group. The other 17 individuals had been wearing fixed orthodontic appliances for at least 3 months each. After a periodontal examination, we collected subgingival plaque samples from the gingivitis lesions of each patient. Using PCR based on 168 rDNA, we detected the presence of 6 putative periodontopathogenic species, Treponema denticola, Porphyromonas gingivalis, Tannerella forsythia (formerly Bacteroides forsythus), Prevotella nigrescens, Prevotella intermedia, and Actinobacillus actinomycetemcomitans. With regard to the presence of individual periodontopathogens, we found that T. forsythia, T. denticola, and P. nigrescens were significantly more common in the samples obtained from the orthodontic patients than in the samples obtained from the non-orthodontic patient controls. Our results indicate that the local changes associated with the wearing of fixed orthodontic appliances may affect the prevalence of periodontopathogens in subgingival dental plaques.

• Restorative Dentistry and Endodontics
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• v.28 no.4
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• pp.303-313
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• 2003

It has been documented that periodontopathic bacteria are also implicated in endodontic infections. 168 rDNA gene-directed PCR was to examine the prevalence of periodontopathic bacteria including Actinobacillus actinomycetemcomitans (Aa), Prevotella intermedia (Pi), Prevotella nigrescens (Pn), Porphyromonas gingivalis (Pg), Porphyromonas endodontalis (Pe), and Treponema denticola (Td) in the root canals of 36 endodontically infected teeth having apical lesions with or without clinical symptoms like pain, swelling, and fistula. 1. In 36 infected root canals, most frequently detected bacterial species was Pg (61.1%), followed by Td (52.8%) and Pe (38.9%). 2. Of 36 infected root canals, Aa was detected in 6 canals (16.7%) of the teeth, all of which showed clinical symptoms. 3. Of 36 infected root canals, Pi and Pn were found in 4 03.9%) and 5 (33.3%), respectively. Notably, prevalence of Pn in the symptomatic teeth was 50.0%. 4. One of black-pigmented anaerobic bacteria (BPB) including Pi, Pn, Pe, and Pg was detected in all of the teeth that showed pain or especially swelling but not fistula. It was, however, found that prevalence of BPB in the asymptomatic teeth or the teeth with fistula was only 40%. 5. Pe and Pg were detected in the teeth regardless of the presence or absence of symptoms. 6. Td was detected in the teeth regardless of the presence or absence of symptoms. High prevalence of BPB in the symptomatic teeth but low in the asymptomatic teeth suggests that BPB may play an important role in the pathogenesis of periapical lesions.

• Journal of Periodontal and Implant Science
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• v.50 no.6
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• pp.358-367
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• 2020

Purpose: The aim of this study was to investigate the efficacy and validity of subgingival bacterial sampling using a retraction cord, and to evaluate how well this sampling method reflected changes in periodontal conditions after periodontal therapy. Methods: Based on clinical examinations, 87 subjects were divided into a healthy group (n=40) and a periodontitis group (n=47). Clinical measurements were obtained from all subjects including periodontal probing depth (PD), bleeding on probing (BOP), clinical attachment loss (CAL), and the plaque index. Saliva and gingival crevicular fluid (GCF) as a subgingival bacterial sample were sampled before and 3 months after periodontal therapy. The salivary and subgingival bacterial samples were analyzed by reverse-transcription polymerase chain reaction to quantify the following 11 periodontal pathogens: Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Tannerella forsythus (Tf), Treponema denticola (Td), Prevotella intermedia (Pi), Fusobacterium nucleatum (Fn), Pavimonas micra (Pm), Campylobacter rectus (Cr), Prevotella nigrescens (Pn), Eikenella corrodens (Ec), and Eubacterium nodatum (En). Results: Non-surgical periodontal therapy resulted in significant decreases in PD (P<0.01), CAL (P<0.01), and BOP (P<0.05) after 3 months. Four species (Pg, Tf, Pi, and Pm) were significantly more abundant in both types of samples in the periodontitis group than in the healthy group. After periodontal therapy, Cr was the only bacterium that showed a statistically significant decrease in saliva, whereas statistically significant decreases in Cr, Pg, and Pn were found in GCF. Conclusions: Salivary and subgingival bacterial sampling with a gingival retraction cord were found to be equivalent in terms of their accuracy for differentiating periodontitis, but GCF reflected changes in bacterial abundance after periodontal therapy more sensitively than saliva.

• Journal of dental hygiene science
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• v.19 no.4
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• pp.213-219
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• 2019

Background: Smoking exerts an adverse effect on the periodontal tissue by reorganizing the ecosystem of oral microorganisms and is considered to be an important factor in the development of periodontal disease. Although cross-sectional studies on smokers and non-smokers have been attempted to investigate the microbial differences in periodontal oral cavity, only few studies have been conducted to investigate the changes in oral microorganisms during smoking cessation. The purpose of this study was to investigate the changes of bacteria in saliva and gingival crevicular fluid (GCF) over a period of one year among 11 smokers trying to quit smoking. Methods: Eleven smokers trying to quit smoking visited the clinic at baseline, two weeks, two months, four months, six months, and 12 months to give saliva and GCF samples. The amounts of 16S rRNA, Porphyromonas gingivalis, Treponema denticola, Prevotella intermedia, Fusobacterium nucleatum subsp. nucleatum, Streptococcus mutans, and Streptococcus sobrinus in saliva and GCF were quantified using real-time polymerase chain reaction TaqMan probe assay. The results were analyzed by nonparametric statistical analysis using Friedman test and Spearman correlation coefficient. Results: After cessation of smoking, the amounts of 16S rRNA corresponding to P. gingivalis, F. nucleatum, P. intermedia, and T. denticola in saliva decreased and then again increased significantly. The amount of F. nucleatum 16S rRNA in GCF decreased significantly after smoking cessation. Positive correlations were observed between 16S rRNA and F. nucleatum and between F. nucleatum and T. denticola in saliva and GCF. Conclusion: Even if the number of subjects in this study was small, we suggest that smoking cessation may reduce the total bacterial amount and F. nucleatum in GCF. However, the results regarding changes in the microbial ecosystem due to smoking or smoking cessation were inconsistent. Therefore, further in-depth studies need to be carried out.

• International Journal of Oral Biology
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• v.39 no.4
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• pp.201-206
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• 2014

This study investigated the antimicrobial activity of methanol extract of mulberry leaf against 16 strains of mutans streptococci and four species of periodontopathogens: Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, and Aggregatibacter actinomycetemcomitans. The antimicrobial activities of the crude extracts or silica gel chromatography fractions of methanol-extracted mulberry leaf were evaluated by determining minimal inhibitory concentrations using an established microdilution method. The cytotoxicity of the extracts of mulberry leaf on KB cells was tested by the methyl thiazolyl tetrazolium assay. Chromatography fraction 12 displayed the most potent antimicrobial activity against all 16 strains of mutans streptococci, P. gingivalis, and P. intermedia. No KB cell cytotoxicity was evident up to $128{\mu}g/ml$ of fraction 12. The methanol extract had no antimicrobial activity against F. nucleatum and A. actinomycetemcomitans. These results suggest chromatography fraction 12 methanol extract of mulberry leaf could be useful in the development of oral hygiene products, such as dentifrice and oral hygiene solution, for the prevention of dental caries.

• Journal of Oral Medicine and Pain
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• v.30 no.1
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• pp.1-14
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• 2005

The maintenance of good oral health in adults is often hindered by oral malodor and periodontal diseases which are known to be commonly caused by some species of Gram-negative anaerobic bacteria, with low sensitivity to common synthetic antibiotics or antibacterial chemical agents. Therefore the development of a nonharmful natural antibacterial oral rinsing remedy against the causative bacteria is thought to be very important. The purpose of this study is to obtain the basic data for development of a nonharmful natural antibacterial oral rinsing remedy using colloidal silver. The author applied colloidal silver solution with concentration of 10, 30, 50, 80 ppm to some strains in species of Prevotella intermedia, Porphyromonas gingivalis, Fusobaterium nucleatum, and evaluated the effects of colloidal silver on the growth of experimental bacterial strains in aspects of minimal inhibitory concentration (MIC), minimal bactericidal concentration (MBC) and growth pattern after incubation for 24, 48, 72 hours. The obtained results were as follows: MIC of colloidal silver solution against experimental strains was 30 ppm in P. intermedia, 10 or 30 ppm in P. gingivalis, and 30, 50, or 80 ppm in F. nucleatum. And MBC of colloidal silver solution against experimental strains was 30 ppm in P. intermedia, 30 or 50 ppm in P. gingivalis, 30 or 80 ppm in F. nucleatum. Therefore it was concluded that colloidal silver exhibited bacteriostatic or/and bacteriocidal effects against some experimental strain. And the inhibition of growth of experimental strains were markedly or considerably exhibited under 30 ppm$\sim$50 ppm of colloidal silver solution for 48 hours$\sim$72 hours in P. intermedia, 10 ppm$\sim$30 ppm for 24 hours$\sim$48 hours in P. gingivalis, 30 ppm for 24 hours in F. nucleatum. These results indicate that the colloidal silver inhibited effectively the growth of some species of Gram-negative anaerobic bacteria by exhibition of bacteriostatic or/and bacteriocidal effects, and can be used as a possible major ingredient of the nonharmful natural antibacterial oral rinsing remedy to oral malodor and periodontal diseases.