• 치위생과학회지
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• 제24권1호
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• pp.54-61
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• 2024

Background: The Korean name for Angelica gigas Nakai (AGN) is Cham-dang-gui, which grows naturally or is cultivated, and its dried roots are used in traditional herbal medicines. The AGN root exert various pharmacological effects. Despite the various pharmacological effects of the AGN root, there are no reports on its anti-oral microbial effects. The purpose of this study was to reveal the anti-oral microbial effect and the microbial and biochemical changes in oral microorganisms according to the concentration of the ethanol extract of AGN (EAGN) root, and to confirm the possibility of using EAGN as a plant-derived functional substance for controlling oral infectious microorganisms. Methods: Disk diffusion test, growth measurement, biofilm formation assay, and measurements of acid production and buffering capacity were performed to confirm the antibacterial effect of EAGN. Results: EAGN showed anti-oral bacterial effects against Streptococcus mutans and Aggregatibacter actinomycetemcomitans at all concentrations, with S. mutans showing a more susceptible effect at concentrations above 5.0 mg/ml and A. actinomycetemcomitans at 3.75 mg/ml. EAGN treatment significantly reduced A. actinomycetemcomitans growth at all concentrations tested. Biofilm formation was significantly reduced at concentrations above 3.75 mg/ml for S. mutans and 2.5 mg/ml for A. actinomycetemcomitans. Acid production in S. mutans and A. actinomycetemcomitans was significantly increased by treatment with EAGN, and the buffering capacities of S. mutans and A. actinomycetemcomitans increased from an EAGN concentration of 3.75 mg/ml and above. Conclusion: EAGN showed anti-oral bacterial effects against both S. mutans and A. actinomycetemcomitans at concentrations above 3.75 mg/ml, which were thought to be related to the inhibition of their growth and biofilm formation. Therefore, EAGN can be used as a safe functional substance derived from medicinal plants owing to its antibacterial effects against S. mutans and A. actinomycetemcomitans.

• 대한소아치과학회지
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• 제44권4호
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• pp.437-445
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• 2017

이 연구의 목적은 한국인들이 일상적으로 마시는 서로 다른 네 종류의 차 티백을 이용하여 차가운 생수나 뜨거운 생수로 5분 또는 10분 동안 우려내는 일상적인 방법으로 차를 추출하였을 때 각각의 추출물이 Streptococcus mutans 세균막 성장에 미치는 효과를 알아보고자 하였다. 균주는 S. mutans UA 159를 이용하였고 1%의 sucrose와 각각의 차 추출물을 웰 플레이트에 넣고 배양하였다. 추출 온도에 따른 세균막 형성을 비교 했을 때 녹차와 홍차에서 추출 온도가 높을 때 세균막 형성이 적었고 통계적으로 유의하였다(p < 0.05). 추출 시간을 달리 하고 $72^{\circ}C$ 온수로 많은 양을 추출 했을 경우 세균막 형성을 비교 했을 때는 네 종류 차 모두 추출 시간에 대해서 통계적으로 유의하지 않았다. 한편 추출량에 따른 비교에서는 녹차와 홍차의 경우 같은 시간, 같은 온도로 추출하였을 때 추출량이 적다면 오히려 세균막 성장이 증가하였다.

• 대한소아치과학회지
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• 제51권1호
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• pp.32-39
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• 2024

This study evaluated the additive impact of ethylenediamine tetraacetic acid (EDTA) on erythrosine-mediated photodynamic therapy (PDT) against Streptococcus mutans (S. mutans) biofilm by measuring colony-forming units and applying confocal laser scanning microscopy. Fifty-six bovine incisors, free from dental caries or structural defects, were utilized in this study. Dentin specimens were created by cutting with a low-speed diamond disk under a continuous flow of water, resulting in dimensions of 6.0 mm × 3.0 mm × 2.0 mm. The specimens were categorized into 4 groups: Control, EDTA, PDT, and EDTA + PDT. S. mutans ATCC 25175 was employed to establish biofilm on the dentin specimens. A 17% EDTA solution was applied for 1 min. For PDT, erythrosine served as the photosensitizer. Finally, a light-emitting diode source (385 - 515 nm) was employed in this study. The PDT group exhibited a significantly lower bacterial count than both the control and EDTA groups (p < 0.001). The EDTA + PDT group demonstrated a significantly reduced bacterial count compared to the other 3 groups (p < 0.001). This study demonstrated that EDTA enhances the antimicrobial efficacy of PDT on S. mutans biofilm. Even at a low concentration of photosensitizer, the combination of EDTA and PDT yields a significant antibacterial effect.

• Journal of Periodontal and Implant Science
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• 제47권4호
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• pp.219-230
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• 2017

Purpose: The purpose of this study was to compare the characteristics of single- and dualspecies in vitro oral biofilms made by static and dynamic methods. Methods: Hydroxyapatite (HA) disks, 12.7 mm in diameter and 3 mm thick, were coated with processed saliva for 4 hours. The disks were divided into a static method group and a dynamic method group. The disks treated with a static method were cultured in 12-well plates, and the disks in the dynamic method group were cultured in a Center for Disease Control and Prevention (CDC) biofilm reactor for 72 hours. In the single- and dual-species biofilms, Fusobacterium nucleatum and Porphyromonas gingivalis were used, and the amount of adhering bacteria, proportions of species, and bacterial reduction of chlorhexidine were examined. Bacterial adhesion was examined with scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). Results: Compared with the biofilms made using the static method, the biofilms made using the dynamic method had significantly lower amounts of adhering and looser bacterial accumulation in SEM and CLSM images. The proportion of P. gingivalis was higher in the dynamic method group than in the static method group; however, the difference was not statistically significant. Furthermore, the biofilm thickness and bacterial reduction by chlorhexidine showed no significant differences between the 2 methods. Conclusions: When used to reproduce periodontal biofilms composed of F. nucleatum and P. gingivalis, the dynamic method (CDC biofilm reactor) formed looser biofilms containing fewer bacteria than the well plate. However, this difference did not influence the thickness of the biofilms or the activity of chlorhexidine. Therefore, both methods are useful for mimicking periodontitis-associated oral biofilms.

• 치위생과학회지
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• 제23권2호
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• pp.103-111
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• 2023

Background: Some studies confirm the reduction of the number of Streptococcus mutans in saliva and dental plaque by Lactobacillus, however, these effects are not always confirmed in in vitro and clinical studies, and only the risk of dental caries has been reported. Our in vitro study aimed to reveal microbial and biochemical changes in the single cultures of S. mutans, Lactobacillus casei and Aggregatibactor actinomycetemcomitans and co-cultures of S. mutans and L. casei or A. actinomycetemcomitans according to sucrose concentration. We also aimed to confirm the anti-oral bacterial and anti-biofilm activities of L. casei and A. actinomycetemcomitans against S. mutans according to sucrose concentration. Methods: S. mutans (KCCM 40105), L. casei (KCCM 12452), and A. actinomycetemcomitans (KCTC 2581) diluted to 5×10⁶ CFU/ml were single cultured, and L. casei or A. actinomycetemcomitans applied at concentrations of 10%, 20%, 30% and 40% to S. mutans were co-cultured with selective medium containing 0%, 1% and 5% sucrose at 36.5℃ for 24 hours. Measurements of bacterial growth value and acid production, disk diffusion and biofilm formation assays were performed. Results: In the medium containing sucrose, the bacterial growth and biofilm formation by S. mutans, L. casei, and A. actinomycetemcomitans were increased. In contrast, 30% and 40% of L. casei in the medium containing 0% sucrose showed both anti-oral bacterial and anti-biofilm activities. This implies that L. casei can be used as probiotic therapy to reduce S. mutans in a 0% sucrose environment. Conclusion: The concentration of sucrose in the oral environment is important for the control of pathogenic bacteria that cause dental caries and periodontitis. To apply probiotic therapy using L. casei for S. mutans reduction, the concentration of sucrose must be considered.

• 생약학회지
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• 제54권1호
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• pp.27-37
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• 2023

This research was conducted to investigate the comprehensive effects of methanol extract of Coptidis rhizoma (MECR) against oral pathogen. We studied the antibacterial, anti-biofilm, anti-gingipain and anti-inflammatory activity of MECR. The minimum bactericidal concentration (MBC) of MECR was 100 ㎍/mL against several oral pathogens. The formation of biofilm of Streptococcus mutans was reduced to 8.93~24.12% in the presence of 25 ㎍/mL of MECR. The gingipain activity of Porphyromonas gingivalis were reduced to 3.91~6.23% in case of Kgp and 5.73~7.78% in case of Rgp in the presence of 10 mg/mL of MECR. The expression of fadA mRNA, virulence factor of Fusobacterium nucleatum (F. nucleatum) was 3 folds decreased in the presence of 25 ㎍/mL of MECR. In case of YD-38 cells challenged with F. nucleatum, RQ values of IL-8 and IL-6 were reduced about 12 folds and 5.45 folds in the presence of 2 ㎍/mL of MECR. In case of RAW 264.7 murine cell challenged with F. nucleatum, RQ values of IL-1β and IL-6 were 2.52 folds and 2.55 folds reduced in the presences of 2 ㎍/mL of MECR. Conclusively, MECR showed potent antibacterial and anti-inflammatory effects against oral pathogenic bacteria.

• 대한치과의사협회지
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• 제58권11호
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• pp.683-689
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• 2020

Dentin surface of non-carious lesion is usually attached with oral biofilm. The biofilm should be removed before application of restorative material, because it may reduce the bond strength of adhesive system. The aim of this study was to evaluate the microtensile bond strength, when the biofilm was removed with brush or bur. Twenty extracted human third molars were sectioned horizontally to obtain dentin surface. Specimen were divided randomly into four group. Biofilm formation was performed in three group, except for Group 1 (negative control). Biofilm was removed as follows: Group 3, using ICB brush; Group 4, using lowspeed round bur #2. Group 2 (positive control) was not removed Biofilm. And in all four groups, the adhesive system (Optibond FL, Kerr) was applied to etched dentin surface, and resin composite was built up in three 1mm increments. After 24 hour storage in distilled water, the teeth were perpendicularly sectioned to obtain beams (1 × 1 mm2). Microtensile bond strength was measured and the data were statistically analyzed using one-way ANOVA and Tukey's post hoc test (p<0.05). Group 4 showed the highest microtensile bond strength (p<0.05), Group 3 showed no significant improvements when compared to Group 1. Group 2 showed lowest microtensile bond strength (p<0.05). When restoring a non-carious cervical lesion, it is essential to remove the biofilm present on the dentin surface. In addition, in the method of removing the biofilm, both the brush removal method and the bur removal method were effective.

• 한국치위생학회지
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• 제24권2호
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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.

• 치위생과학회지
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• 제16권4호
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• pp.284-292
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• 2016

DUWL에 형성된 바이오필름 제거를 위한 효과적인 소독제의 제시와 새로운 소독제의 개발을 위해 DUWL의 실험실 모델의 확립이 필요하다. 따라서 본 연구에서는 실험실에서 쉽게 구할 수 있는 장비들로 실험실 모델을 제작하여, DUWL 바이오필름을 재현하기 위한 새로운 실험실 모델을 확립하고자 하였다. 사용 중인 DUWL을 통해 수집한 물에서 세균을 모은 후, R2A 액체 배지에서 10일 동안 배양시켰다. 10일 배양시킨 세균액을 $-70^{\circ}C$에 보관하여 사용하였다. $-70^{\circ}C$에 저장한 세균 stock은 R2A 액체배지에 5일 동안 회분 배양시킨 배양액은 모델에서 바이오필름을 형성시키기 위해 사용되었다. 바이오필름 형성 모델은 실험실 내 장비인 1 L 비커에 폴리우레탄 튜빙이 부착된 20 cm 유리막대를 꽂아서 제작하였다. 모델을 멸균시킨 후 R2A 액체배지 300 ml와 5일 동안 회분 배양한 세균액 50 ml을 넣고 stir plate에서 $25^{\circ}C$로 배양시켰다. 배양 2일마다 R2A 액체배지를 교체해주었다. 임상의 상황과 유사한 조건에서 바이오필름을 형성하기 위해 와류상태는 오전 9시에서 오후 6시까지 적용시키고 그 이외의 시간에는(약 15시간) 정체상태로 배양시켰다. 바이오필름 형성은 4일 동안 진행하였으며, 그 후 바이오필름의 두께, 바이오필름을 구성하는 세균의 분포 및 형태학적 특징을 SEM과 CLSM을 사용하여 분석하였다. 4일 바이오필름 형성 후 평균 바이오필름 축적량은 $4.68{\times}10^4CFU/cm^2$였고, 바이오필름의 두께는 $10{\sim}14{\mu}m$였다. 또한 바이오필름을 구성하는 세균들이 부분적으로 응집되어 덩어리를 이루고 있는 양상을 확인할 수 있었다. 본 연구에서 제작한 실험실 모델을 대상으로 차아염소산나트륨, 과산화수소 그리고 클로르헥시딘과 같은 소독제의 효과를 확인하였다. 그 결과 적용된 소독제의 농도가 낮을수록 바이오필름 내 생존한 세균의 수가 많았다. 따라서 우리의 실험실 모델에서 형성시킨 바이오필름은 소독제의 효과를 비교하기 위해 적절한 것으로 판단된다. 우리의 실험실 모델은 향후 DUWL 소독을 위한 새로운 방법의 개발을 위해 유용하게 사용될 것으로 예상된다.

• 대한소아치과학회지
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• 제49권2호
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• pp.149-157
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• 2022

이 연구의 목적은 erythrosine 매개 광역동 치료 (Photodynamic therapy, PDT)가 Streptococcus mutans 바이오필름 회복에 미치는 영향을 Colony Forming Unit (CFU) 측정과 공초점 레이저 주사 현미경의 관찰을 통해 평가하는 것이었다. PDT에서 광감각제로 erythrosine을 사용하였다. S. mutans ATCC 25175 바이오필름에 LED 광원을 통해 광역동 치료를 시행하였다. 클로르헥시딘 (Chlorhexidine, CHX) 처리한 군을 양성대조군으로 설정하였다. 각 군에 따른 처리 후 바이오필름의 회복을 위해 배양하였다. 세균 생존율을 처리 직후, 재배양 후 3시간 간격으로 24시간까지 CFU 계수를 통해 측정하였다. 항균 처리 직후 PDT, CHX에서 모두 음성대조군과 비교 시 S. mutans CFU 수가 유의하게 감소하였다. 재배양 12시간 후 PDT는 음성대조군과 비교 시 CFU 수 감소에 관하여 통계적인 유의성을 띄지 않았지만 CHX는 24시간 동안 통계적으로 낮은 CFU 수를 보였다. Erythrosine을 이용한 광역동 치료는 S. mutans 바이오필름 형성을 효과적으로 억제하지만 클로르헥시딘보다 바이오필름의 회복이 빠르게 나타났다. 이 연구는 치아 우식 예방을 위한 광역동 치료의 임상적 효과에 관한 통찰력을 제공한다.