• 한국축산식품학회지
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• 제42권6호
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• pp.1020-1030
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• 2022

The aim of the study was to investigate the effect of bacteriocin-like inhibitory substance (BLIS) from Enterococcus faecium DB1 on cariogenic Streptococcus mutans biofilm. Crystal violet staining, fluorescence, and scanning electron microscopy analyses demonstrated that the BLIS from Enterococcus faecium DB1 (DB1 BLIS) inhibited S. mutans biofilm. When DB1 BLIS was co-incubated with S. mutans, biofilm formation by S. mutans was significantly reduced (p<0.05). DB1 BLIS also destroyed the preformed biofilm of S. mutans. In addition, DB1 BLIS decreased the viability of S. mutans biofilm cells during the development of biofilm formation and in the preformed biofilm. DB1 BLIS significantly decreased the growth of S. mutans planktonic cells. Furthermore, S. mutans biofilm on the surface of saliva-coated hydroxyapatite discs was reduced by DB1 BLIS. Taken together, DB1 BLIS might be useful as a preventive and therapeutic agent against dental caries caused by S. mutans.

• Journal of Microbiology and Biotechnology
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• 제26권11호
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• pp.1829-1835
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• 2016

Dental caries is caused by cariogenic biofilm, an oral biofilm including Streptococcus mutans. Recently, the prevention of dental caries using various probiotics has been attempted. Lactococcus lactis HY 449 is a probiotic bacterium. The aim of this study was to investigate the effect of L. lactis HY 449 on cariogenic biofilm and to analyze its inhibitory mechanisms. Cariogenic biofilm was formed in the presence or absence of L. lactis HY 449 and L. lactis ATCC 19435, and analyzed with a confocal laser microscope. The formation of cariogenic biofilm was reduced in cultures spiked with both L. lactis strains, and L. lactis HY 449 exhibited more inhibitory effects than L. lactis ATCC 19435. In order to analyze and to compare the inhibitory mechanisms, the antibacterial activity of the spent culture medium from both L. lactis strains against S. mutans was investigated, and the expression of glucosyltransferases (gtfs) of S. mutans was then analyzed by real-time RT-PCR. In addition, the sucrose fermentation ability of both L. lactis strains was examined. Both L. lactis strains showed antibacterial activity and inhibited the expression of gtfs, a nd t he d ifference b etween both strains did not show. In the case of sucrose-fermenting ability, L. lactis HY 449 fermented sucrose but L. lactis ATCC 19435 did not. L. lactis HY 449 inhibited the uptake of sucrose and the gtfs expression of S. mutans, whereby the development of cariogenic biofilm may be inhibited. In conclusion, L. lactis HY 449 may be a useful probiotic for the prevention of dental caries.

• Journal of Microbiology and Biotechnology
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• 제27권7호
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• pp.1242-1248
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• 2017

Several medicinal plants are ethnomedically used in Korea as agents for treating infection, anti-inflammation, and pain relief. However, beyond typical inhibitory effects on cell growth, little is known about the potential anti-biofilm activity of these herbs, which may help to prevent cavities and maintain good oral health. This study aimed to investigate the antimicrobial and anti-biofilm activities of the methanol extracts of 37 Korean medicinal plants against dental pathogens Streptococcus mutans and Candida albicans, which synergize their virulence so as to induce the formation of plaque biofilms in the oral cavity. The antimicrobial activities were investigated by broth dilution and disk diffusion assay. The anti-biofilm and antioxidant activities were evaluated based on the inhibitory effect against glucosyltransferase (GTase) and the DPPH assay, respectively. Among 37 herbs, eight plant extracts presented growth and biofilm inhibitory activities against both etiologic bacteria. Among them, the methanol extracts (1.0 mg/ml) from Camellia japonica and Thuja orientalis significantly inhibited the growth of both bacteria by over 76% and over 83% in liquid media, respectively. Minimum inhibitory concentration (MIC) values of these methanol extracts were determined to be 0.5 mg/ml using a disk diffusion assay on solid agar media. Biofilm formation was inhibited by more than 92.4% and 98.0%, respectively, using the same concentration of each extract. The present results demonstrate that the medicinal plants C. japonica and T. orientalis are potentially useful as antimicrobial and anti-biofilm agents in preventing dental diseases.

• 치위생과학회지
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• 제18권2호
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• pp.69-75
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• 2018

우리 연구의 목적은 DUWL에서 배출되는 물에서 분리한 균주의 부착 능력과 바이오필름 형성 능력을 확인하고 두 능력 사이 관계를 확인하는 것이다. DUWL로부터 분리한 12균주를 실험에 사용하였다. 각 균주의 부착 능력을 확인하기 위해, 12-well plates의 각 well에 멸균된 glass coverslip, R2A 액체 배지, 그리고 $1{\times}10^8CFU/ml$의 농도로 조정된 세균 현탁액을 넣고 $26^{\circ}C$ 배양기에서 7일 동안 배양하였다. 배양 후, glass coverslip에 부착한 정도에 따라 1~3점으로 점수를 부여하였다. 분리 균주의 바이오필름 형성 능력을 확인하기 위해, 96-well polystyrene flat-bottom microtiter plate에 R2A 액체 배지와 세균 현탁액을 넣고 $26^{\circ}C$에서 7일 동안 배양하였다. 배양 후, plate에 형성된 바이오필름은 R2A 액체 배지에 현탁했고, 현탁액을 R2A 고체 배지에 도말하였다. $26^{\circ}C$에서 7일 배양한 후 세균의 집락을 계수하고 CFU/ml를 계산하였다. DUWL로부터 총 56균주가 분리되었으며, 12속과 31종을 포함하였다. 실험에는 속당 1균주씩 선택하여 총 12균주가 사용되었다. 12균주 중에 S. echinoides, M. aquaticum, C. pauculus의 부착 능력 점수는 +3으로 가장 높았다. 바이오필름 축적량은 C. pauculus가 가장 많았고, M. testaceum이 가장 적었다. 대부분의 부착 능력이 높은 균주는 바이오필름 형성 능력 또한 높았다. 본 연구의 결과는 DUWL 바이오필름의 형성 기전을 파악하는데 도움을 주며 나아가 바이오필름 형성을 억제하는 방법의 개발에 기본적인 정보를 제공할 수 있을 것이다.

• 대한치과교정학회지
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• 제53권6호
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• pp.345-357
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• 2023

Enamel demineralization represents the most prevalent complication arising from fixed orthodontic treatment. Its main etiology is the development of cariogenic biofilms formed around orthodontic appliances. Ordinarily, oral biofilms exist in a dynamic equilibrium with the host's defense mechanisms. However, the equilibrium can be disrupted by environmental changes, such as the introduction of a fixed orthodontic appliance, resulting in a shift in the biofilm's microbial composition from non-pathogenic to pathogenic. This alteration leads to an increased prevalence of cariogenic bacteria, notably mutans streptococci, within the biofilm. This article examines the relationships between oral biofilms and orthodontic appliances, with a particular focus on strategies for effectively managing oral biofilms to mitigate enamel demineralization around orthodontic appliances.

• International Journal of Oral Biology
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• 제41권4호
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• pp.253-262
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• 2016

Streptococcus mutans (S. mutans) is one of the most important bacteria in the formation of dental plaque and dental caries. S. mutans adheres to an acquired pellicle formed on the tooth surface, and aggregates with many oral bacteria. It initiates plaque formation by synthesizing glucan from sucrose, which is catalyzed by glucosyltransferases. Propolis is a resinous mixture produced by honeybees, by mixing saliva and beeswax with secretions gathered from wood sap and flower pollen. Bees prevent pathogenic invasions by coating the propolis to the outer and inner surface of the honeycomb. Propolis has traditionally been used for the treatment of allergic rhinitis, asthma and dermatitis. We investigated the inhibitory effects of propolis ethanol extract on biofilm formation and gene expression of S. mutans. The biofilm formation of S. mutans was determined by scanning electron microscopy (SEM) and safranin staining. We observed that the extract of propolis had an inhibitory effect on the formation of S. mutans biofilms at concentrations higher than 0.2 mg/ml. Real-time PCR analysis showed that the gene expression of biofilm formation, such as gbpB, spaP, brpA, relA and vicR of S. mutans, was significantly decreased in a dose dependent manner. The ethanol extract of propolis showed concentration dependent growth inhibition of S. mutans, and significant inhibition of acid production at concentrations of 0.025, 0.05, 0.1 and 0.2 mg/ml, compared to the control group. These results suggest that the ethanol extract of propolis inhibits gene expression related to biofilm formation in S. mutans.

• Journal of Periodontal and Implant Science
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• 제48권1호
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• pp.12-21
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• 2018

Purpose: The goal of this study was to develop and validate a standardized in vitro pathogenic biofilm attached onto saliva-coated surfaces. Methods: Fusobacterium nucleatum (F. nucleatum) and Porphyromonas gingivalis (P. gingivalis) strains were grown under anaerobic conditions as single species and in dual-species cultures. Initially, the bacterial biomass was evaluated at 24 and 48 hours to determine the optimal timing for the adhesion phase onto saliva-coated polystyrene surfaces. Thereafter, biofilm development was assessed over time by crystal violet staining and scanning electron microscopy. Results: The data showed no significant difference in the overall biomass after 48 hours for P. gingivalis in single- and dual-species conditions. After adhesion, P. gingivalis in single- and dual-species biofilms accumulated a substantially higher biomass after 7 days of incubation than after 3 days, but no significant difference was found between 5 and 7 days. Although the biomass of the F. nucleatum biofilm was higher at 3 days, no difference was found at 3, 5, or 7 days of incubation. Conclusions: Polystyrene substrates from well plates work as a standard surface and provide reproducible results for in vitro biofilm models. Our biofilm model could serve as a reference point for studies investigating biofilms on different surfaces.

• The Journal of Advanced Prosthodontics
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• 제5권2호
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• pp.140-146
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• 2013

PURPOSE. The aim of this study was to investigate the destructive effects of biofilm formation and/or biocorrosive activity of 6 different oral microorganisms. MATERIALS AND METHODS. Three different heat polymerized acrylic resins (Ivocap Plus, Lucitone 550, QC 20) were used to prepare three different types of samples. Type "A" samples with "V" type notch was used to measure the fracture strength, "B" type to evaluate the surfaces with scanning electron microscopy and "C" type for quantitative biofilm assay. Development and calculation of biofilm covered surfaces on denture base materials were accomplished by SEM and quantitative biofilm assay. According to normality assumptions ANOVA or Kruskal-Wallis was selected for statistical analysis (${\alpha}$=0.05). RESULTS. Significant differences were obtained among the adhesion potential of 6 different microorganisms and there were significant differences among their adhesion onto 3 different denture base materials. Compared to the control groups after contamination with the microorganisms, the three point bending test values of denture base materials decreased significantly (P<.05); microorganisms diffused at least 52% of the denture base surface. The highest median quantitative biofilm value within all the denture base materials was obtained with P. aeruginosa on Lucitone 550. The type of denture base material did not alter the diffusion potential of the microorganisms significantly (P>.05). CONCLUSION. All the tested microorganisms had destructive effect over the structure and composition of the denture base materials.

• Journal of Microbiology
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• 제45권4호
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• pp.291-296
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• 2007

Insoluble glucans synthesized by Streptococcus mutans enhance the pathogenicity of oral biofilm by promoting the adherence and accumulation of cariogenic bacteria on the surface of the tooth. The objective of this study was to investigate the effect of Leuconostoc spp. on the in vitro formation of S. mutans biofilm. Three strains, Leuconostoc gelidum A TCC 49366, Leuconostoc mesenteroides ssp. cremoris A TCC 19254 and Leuconostoc mesenteroides ssp. mesenteroides ATCC 8293, were used in this study. They exhibited profound inhibitory effects on the formation of S. mutans biofilm and on the proliferation of S. mutans. The water-soluble polymers produced from sucrose were most strongly produced by L. gelidum, followed by L. mesenteroides ssp. cremoris and L. mesenteroides ssp. mesenteroides. The mean wet weights of the artificial biofilm of S. mutans were also significantly reduced as a result of the addition of the water-soluble polymers obtained from Leuconostoc cultures. According to the results of thin-layer chromatographic analysis, the hydrolysates of the water-soluble polymers produced by Leuconostoc were identical to those of dextran T-2000, forming predominately ${\alpha}-(1-6)$ glucose linkages. These results indicate that dextran-producing Leuconostoc strains are able to inhibit the formation of S. mutans biofilm in vitro.