• Journal of Periodontal and Implant Science
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• 제45권1호
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• pp.14-22
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• 2015

Purpose: The purpose of this study was to compare serum amyloid A (SAA) protein levels with high-sensitive C-reactive protein (hs-CRP) levels as markers of systemic inflammation in patients with chronic periodontitis. The association of serum titers of antibodies to periodontal microbiota and SAA/hs-CRP levels in periodontitis patients was also studied. Methods: A total of 110 individuals were included in this study. Patients were assessed for levels of hs-CRP and SAA. Nonfasting blood samples were collected from participants at the time of clinical examination. The diagnosis of adipose tissue disorders was made according to previously defined criteria. To determine SAA levels, a sandwich enzyme-linked immunosorbent assay was utilized. Paper points were transferred to a sterile tube to obtain a pool of samples for polymerase chain reaction processing and the identification of Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, and Tannerella forsythia. The serum level of IgG1 and IgG2 antibodies to P. gingivalis, A. actinomycetemcomitans, and T. forsythia was also determined. Results: SAA and hs-CRP levels were higher in periodontitis patients than in controls (P<0.05). In bivariate analysis, high levels of hs-CRP (>3 mg/L) and SAA (>10 mg/L) were significantly associated with chronic periodontitis (P=0.004). The Spearman correlation analysis between acute-phase proteins showed that SAA positively correlated with hs-CRP (r=0.218, P=0.02). In the adjusted model, chronic periodontitis was associated with high levels of SAA (odds ratio [OR], 5.5; 95% confidence interval [CI], 1.6-18.2; P=0.005) and elevated hs-CRP levels (OR, 6.1, 95% CI, 1.6-23.6; P=0.008). Increased levels of serum IgG2 antibodies to P. gingivalis were associated with high levels of SAA (OR, 3.6; 95% CI, 1.4-8.5; P=0.005) and high concentrations of hs-CRP (OR, 4.3; 95% CI, 1.9-9.8; P<0.001). Conclusions: SAA and hs-CRP concentrations in patients with chronic periodontitis are comparably elevated. High serum titers of antibodies to P. gingivalis and the presence of periodontal disease are independently related to high SAA and hs-CRP levels.

• International Journal of Oral Biology
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• 제37권4호
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• pp.197-201
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• 2012

LIVE/DEAD$^{(R)}$ BacLight$^{TM}$ and alamarBlue$^{(R)}$ are fluorescent materials used for the enumeration of live and dead bacteria. LIVE/DEAD$^{(R)}$ BacLight$^{TM}$ is generally used for confocal microscopy applications to differentiate live from dead bacteria in a biofilm or planktonic state. AlamarBlue$^{(R)}$ has also been used widely to assay live and dead bacteria in a planktonic state. Whilst these materials are successfully utilized in experiments to discriminate live from dead bacteria for several species of bacteria, the application of these techniques to oral bacteria is limited to the use of LIVE/DEAD$^{(R)}$ BacLight$^{TM}$ in biofilm studies. In our present study, we assessed whether these two methods could enumerate live and dead oral bacterial species in a planktonic state. We tested the reagents on Streptococcus mutans, Streptococcus sobrinus, Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans and Enterococcus faecalis and found that only LIVE/DEAD$^{(R)}$ BacLight$^{TM}$ could differentiate live from dead cells for all five of these oral strains. AlamarBlue$^{(R)}$ was not effective in this regard for P. gingivalis or A. actinomycetemcomitans. In addition, the differentiation of live and dead bacterial cells by alamarBlue$^{(R)}$ could not be performed for concentrations lower than $2{\times}10^6$ cells/ml. Our data thus indicate that LIVE/DEAD$^{(R)}$ BacLight$^{TM}$ is a more effective reagent for this analysis.

• Journal of Periodontal and Implant Science
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• 제51권6호
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• pp.409-421
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• 2021

Purpose: The aim of this study was to compare the prevalence and bacterial load of 6 main periodontal pathogens between pairs of periodontal patients with and without type 2 diabetes mellitus. Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans genotypes were also investigated. Methods: Twenty patients affected by chronic periodontitis and type 2 diabetes were retrospectively selected and matched to 20 patients without diabetes on the basis of the degree and severity of periodontal disease. Microbiological data of subgingival biofilms were analysed and compared for the examined pathogens: A. actinomycetemcomitans, P. gingivalis, Prevotella intermedia, Treponema denticola, Fusobacterium nucleatum, and Tannerella forsythia. Results: The pairs were balanced in terms of demographic and clinical parameters, except for bleeding on probing and suppuration. In the microbiological test sites (4 for each patient), the mean probing pocket depth was 6.34±1.63 mm in patients with diabetes and 6.41±1.78 mm in patients without diabetes. No significant difference between pairs in the prevalence of P. gingivalis or the distribution of its genotypes was recorded. Patients with diabetes had a significantly greater amount of total bacterial load, P. gingivalis, T. denticola, T. forsythia, and F. nucleatum (P<0.05). Moreover, patients with diabetes had a higher number of sites with a greater cell count than patients without diabetes. When compared to the total bacterial load, only T. forsythia maintained its relative load in patients with diabetes (P=0.001). Conclusions: This retrospective matched study supports the hypothesis that microbiological differences exist among periodontal patients with and without diabetes mellitus.

• 대한소아치과학회지
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• 제35권3호
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• pp.494-503
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• 2008

Real-time PCR 기법을 이용하여 구강내 치태에서 특정 세균의 존재여부를 알 수 있을 뿐 아니라 정량적 분석이 가능하게 되었다. 이 연구는 real-time PCR 기법을 이용하여 8-18세의 청소년에서 치주질환 원인균의 조성을 알아보고자 전신질환이 없는 어린이 및 청소년 65명을 대상으로 치은연하의 치태를 채취하여 5종의 치주질환 원인균의 출현율과 그 양을 측정하고, 또한 함께 측정한 치태지수 및 치은지수와의 상관관계를 조사하여 다음과 같은 결론을 얻었다. 1. 치태지수의 평균은 1.33이었으며, 치은지수의 평균은 0.97이었다. 치태지수는 나이가 증가할수록 낮아졌다(p<0.05). 2. 치주질환 원인균의 출현율은 P. gingivalis 61.5%, T. forsythia 53.8%, T. denticola 29.2%, A. actinomycetemcomitans 15.4%, F. nucleatum은 100%였다. P. gingivalis, T. denticola는 10세 이후 출현율이 증가하였고 A. actinomycetemcomitans는 8-10세 군에서 출현율이 높았다(p<0.05). 3. 정량적 분석에서 치태지수와 치은지수는 F. nucleatum의 양과 유의한 상관성이 있었으며, 치태지수는 치은지수와 유의한 상관성이 있었다. T. forsythia는 A. actinomycetemcomitans, T. denticola양과 강한 상관성을 보였으며, T. denticola양과 A. actinomycetemcomitans도 강한 상관관계를 가지고 있었다.

• Restorative Dentistry and Endodontics
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• 제28권2호
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• pp.178-183
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• 2003

The purpose of this study was to investigate the frequency of 7 putative pathogens in endodontic infections. The specimens were collected from infected pulpal tissue of patients who were referred for root canal treatment to the department of conservative dentistry, Chosun University Samples were collected aseptically using a barbed broach and a paper point. The cut barbed broaches and paper points were transferred to an eppendorf tube containing 500 ml of 1 X PBS. DNAs were extracted from the samples by direct DNA extraction method using lysis buffer (0.5% EDTA, 1% Triton X-100). Identification of 7 putative pathogens was performed by PCR based on 16S rDNA. The target species were as follows : Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Bacteroides forsythus, Actinobacillus actinomycetemcomitans, and Treponema denticola. Our data revealed that the prevalence of P. endodontalis was found in 88.6% (39/54), P. ginivalis 52.3% (23/44), P. nigrescens 18.2% (8/44), P intermedia 15.9% (7/44) B. forsythus 18.2% (8/44), A. actinomycetemcomitans 3.3% (1/44), T. denticola 25% (l1/44) of the samples. The high prevalence of P. endodontalis and P. ginivalis suggests that they may play an important role in the etiology of endodontic infections.

• International Journal of Oral Biology
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• 제38권4호
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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 Periodontal and Implant Science
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• 제42권6호
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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.

• Journal of Periodontal and Implant Science
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• 제53권2호
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• pp.110-119
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• 2023

Purpose: This study aimed to investigate the proper wavelengths for safe levels of light-emitting diode (LED) irradiation with bactericidal and photobiomodulation effects in vitro. Methods: Cell viability tests of fibroblasts and osteoblasts after LED irradiation at 470, 525, 590, 630, and 850 nm were performed using the thiazolyl blue tetrazolium bromide assay. The bactericidal effect of 470-nm LED irradiation was analyzed with Streptococcus gordonii, Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis, and Tannerella forsythia. Levels of nitric oxide, a proinflammatory mediator, were measured to identify the anti-inflammatory effect of LED irradiation on lipopolysaccharide-stimulated inflammation in RAW 264.7 macrophages. Results: LED irradiation at wavelengths of 470, 525, 590, 630, and 850 nm showed no cytotoxic effect on fibroblasts and osteoblasts. LED irradiation at 630 and 850 nm led to fibroblast proliferation compared to no LED irradiation. LED irradiation at 470 nm resulted in bactericidal effects on S. gordonii, A. actinomycetemcomitans, F. nucleatum, P. gingivalis, and T. forsythia. Lipopolysaccharide (LPS)-induced RAW 264.7 inflammation was reduced by irradiation with 525-nm LED before LPS treatment and irradiation with 630-nm LED after LPS treatment; however, the effects were limited. Conclusions: LED irradiation at 470 nm showed bactericidal effects, while LED irradiation at 525 and 630 nm showed preventive and treatment effects on LPS-induced RAW 264.7 inflammation. The application of LED irradiation has potential as an adjuvant in periodontal therapy, although further investigations should be performed in vivo.

• International Journal of Oral Biology
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• 제39권4호
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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.

• International Journal of Oral Biology
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• 제32권2호
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• pp.75-78
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• 2007

The periodontal diseases and dental caries are major infectious diseases in oral cavity. Many of the preventive and therapeutic dental products contain the antimicrobial and anti-inflammatory components. But some of these antimicrobial have weak points such as coloration, burning sensation and insolubility in water. We have screened the therapeutic herbal extracts of the Plant Extract Bank for the antimicrobial activity on the major dental pathogens by growth inhibition assay. For the Porphyromonas gingivalis, 8 herbal extracts had an antimicrobial activity, 11 herbal extracts for the Prevotella intermedia, 43 herbal extracts for the Haemophilus actinomycetemcomitans and 61 herbal extracts for the Streptococcus mutans. Among these extracts, 6 herbal extracts had an antimicrobial activity for more than 3 species of dental pathogens. These extracts are Araliae Cordatae Radix, Crassirhizomae Rhizoma, Mori Radicis Cortex, Psoraleae Semen, Pini Ramulus and Sieges- beckiae Herba. All of effective extracts were CA group, ethanol extracts. Among these 6 herbal extracts, only Crassirhizomae is known to have a antibacterial effects. Therefore these herbal extracts have a possibility to be a candidate for a major antibacterial components in dental products.