• International Journal of Oral Biology
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• 제46권1호
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• pp.60-65
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• 2021

Dental health care workers (DHCW) are at a risk of occupational exposure to Helicobacter pylori from the aerosolized oral biofilms and saliva of patients. We designed this study to investigate the prevalence of H. pylori in the oral biofilms of a group of dental and non-dental undergraduates from Sri Lanka. After obtaining informed consent, oral biofilms were collected from 38 dental undergraduates (19 males and 19 females) undergoing clinical training and 33 non-dental undergraduates (14 males and 19 females). The participants were in the age range of 22-27 years and had healthy periodontium. Total DNA from the oral biofilms were extracted, and H. pylori DNA was detected using polymerase chain reaction (PCR) amplification of 16S rRNA gene of H. pylori using JW22-JW23 primers, and the results were confirmed using PCR amplification of H. pylori-urease specific HPU1-HPU2 primers. Out of 71 participants, 11 (28.95%) dental and 3 (9.09%) non-dental undergraduates had H. pylori in their oral biofilms indicating an overall prevalence rate of 19.72% (14/71). Thus, the prevalence of H. pylori in oral biofilms was significantly higher in dental undergraduates than in non-dental undergraduates (p < 0.05). An odds ratio of 4.07 indicated that dental undergraduates were four times more likely to harbor H. pylori in their oral biofilms than non-dental undergraduates. Foregoing data support the fact that there may be greater occupational risk of exposure to H. pylori for dental undergraduates during clinical training than that for non-dental undergraduates, warranting meticulous infection control practices during clinical dentistry.

• 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.

• Food Science and Biotechnology
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• 제18권2호
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• pp.556-560
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• 2009

Xanthorrhizol, a sesquiterpene isolated from Curcuma xanthorrhiza Roxb., was used to investigate its effect on reducing the saliva and multi-species oral biofilms consisting of Streptococcus mutans, Streptococcus sanguis, and Actinomyces viscosus by anti-biofilm and confocal laser scanning microscopy (CLSM) assays. Xanthorrhizol exhibited significant antibiofilm activity in the dose- and time-dependent manners. Exposure to 2 and $5{\mu}g/mL$ xanthorrhizol for 30 min remained <50% of saliva and multi-species biofilms formed for 24 hr. In addition, exposure to $10{\mu}g/mL$ xanthorrhizol for 30 min reduced 65 and 77% of 24 hr saliva and multi-species oral biofilms, respectively. CLSM results visually demonstrated that xanthorrhizol reduced bacterial viability in the saliva and multi-species oral biofilms. These results suggest that C. xanthorrhiza Roxb. containing xanthorrhizol with strong anti-biofilm activity can be employed as a plant source for oral care functional foods.

• Journal of Periodontal and Implant Science
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• 제44권2호
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• pp.79-84
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• 2014

Purpose: While single-species biofilms have been studied extensively, we know notably little regarding multispecies biofilms and their interactions. The purpose of this study was to develop and evaluate an in vitro multispecies dental biofilm model that aimed to mimic the environment of chronic periodontitis. Methods: Streptococcus gordonii KN1, Fusobacterium nucleatum ATCC23726, Aggregatibacter actinomycetemcomitans ATCC33384, and Porphyromonas gingivalis ATCC33277 were used for this experiment. The biofilms were grown on 12-well plates with a round glass slip (12 mm in diameter) with a supply of fresh medium. Four different single-species biofilms and multispecies biofilms with the four bacterial strains listed above were prepared. The biofilms were examined with a confocal laser scanning microscope (CLSM) and scanning electron microscopy (SEM). The minimum inhibitory concentrations (MIC) for four different planktonic single-species and multispecies bacteria were determined. The MICs of doxycycline and chlorhexidine for four different single-species biofilms and a multispecies biofilm were also determined. Results: The CLSM and SEM examination revealed that the growth pattern of the multispecies biofilm was similar to those of single-species biofilms. However, the multispecies biofilm became thicker than the single-species biofilms, and networks between bacteria were formed. The MICs of doxycycline and chlorhexidine were higher in the biofilm state than in the planktonic bacteria. The MIC of doxycycline for the multispecies biofilm was higher than were those for the single-species biofilms of P. gingivalis, F. nucleatum, or A. actinomycetemcomitans. The MIC of chlorhexidine for the multispecies biofilm was higher than were those for the single-species biofilms of P. gingivalis or F. nucleatum. Conclusions: To mimic the natural dental biofilm, a multispecies biofilm composed of four bacterial species was grown. The 24-hour multispecies biofilm may be useful as a laboratory dental biofilm model system.

• 한국치위생학회지
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• 제23권4호
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• pp.209-218
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• 2023

연구목적: 실험실내에서 성분이 다른 구강세정제가 오래 형성된 다중 구강 바이오필름에 미치는 항균효과를 확인하였다. 연구방법: 구강세정제의 항균효과를 ATP, CFU로 확인하였고, 다중 구강 바이오필름을 형성시킨 sHA 디스크를 구강세정제로 처리한 후 SEM을 이용하여 형태학적 모양을 관찰하였다. 총RNA는 P. intermedia의 바이오필름에서 추출하였고 RNA sequencing 분석을 하였다. 연구결과: 대조군과 구강세정제 4개를 비교한 CFU 측정 결과에서 오래 형성된 다중 구강 바이오필름은 39.0%로부터 95.7%까지 감소한 것으로 나타났다(p<0.05). 에센셜 오일 처리 후 구강생물막 세포들은 크기가 변하고 일부 파열되고 작은 세포 파편 조각도 나타났다. 에센셜 오일로 처리 후 P. intermedia 유전자 발현은 RNA 전사와 단백질 번역 관련 유전자에서 유의한 변화를 보였다. 결론: 성분이 다른 구강세정제는 항균효과를 가지며 표면 구조 및 유전자 발현에 영향을 미친다.

• 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.

• International Journal of Oral Biology
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• 제35권4호
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• pp.197-202
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• 2010

Most oral microorganisms exist as biofilms which initiate formation via the attachment of an early colonizer to host proteins on the tooth surface. Fusobacterium nucleatum act as a bridge between early and late colonizers. Dental biofilms eventually comprise dental pathogens such as Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia. To evaluate the effects of mutual interactions between oral bacteria on the growth of biofilms, periodontopathogens were co-cultured with a $0.4\;{\mu}m$ barrier. Streptococcus gordonii inhibited the growth of F. nucleatum and periodontopathogens. However, F. nucleatum, P. gingivalis and T. denticola activated the growth of other bacteria. A co-culture system of early and late colonizers could be a useful tool to further understand bacterial interactions during the development of dental biofilm.

• Journal of Korean Academy of Oral Health
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• 제43권3호
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• pp.118-123
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• 2019

Objectives: The purpose of this study was to investigate changes in the composition of artificial cariogenic biofilms using a Streptococcus mutans biofilm model over a period of time. Methods: We analyzed the dry weight, colony forming unit (CFU) number, extracellular polysaccharide (EPS) biovolume, and acid production rate of S. mutans biofilms formed on saliva-coated hydroxyapatite discs after 26 h, 50 h, 74 h, 98 h, 171 h, and 195 h. In addition, we performed a laser scanning confocal fluorescence microscopy to determine the bacterial volume, EPS biovolume, and biofilm thickness. We calculated the biofilm density using dry weight and EPS biovolume. Results: Over a period of time, there was no change in the CFU number and acid production rate of S. mutans biofilms, but there was an increase in the dry weight and EPS biovolume of S. mutans biofilms. The bacterial volume, EPS biovolume, and biofilm thickness only increased in the 50-h-old biofilm; however, no change was observed in 50-195-h-old biofilms. In addition, an increase in the biofilm density was observed over time. Conclusions: These results suggest that the acid production ability of cariogenic biofilms does not change, but the biofilm density increases over time. However, due to scientific information, further research needs to be conducted in the field of dentistry to get further insights on the progression of cariogenic biofilms over time.

• 생명과학회지
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• 제31권1호
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• pp.100-108
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• 2021

구강에는 700여 종의 미생물이 부유성 세균(planktonic cells)으로 있거나 치아 표면에 부착하여 바이오필름(biofilms)을 형성하고 있으며 구강 바이오필름(oral biofilms)과 관련된 대표적인 구강질환에는 치아우식증과 치주질환이 있다. 구강 세균은 타액의 흐름, 숙주의 항균 단백질, 영양소의 가용성 및 pH 변화, 항생제, 방부제 등의 영향을 받고 있지만 바이오필름이 형성되면 바이오필름은 물리적으로 두꺼운 층을 이루고 있기 때문에 구강 세균은 이와 같은 구강 내부의 환경적 스트레스에 저항할 수 있을 뿐만 아니라 외부 물질의 침투가 어렵고 바이오필름 내 세포간 상호작용을 통해 유전자 변이가 일어나기 때문에 부유성 세균보다 항생제에 대한 저항성이 1,000배 정도 높다. 따라서 구강 세균의 바이오필름 형성을 억제하거나 제거하면 보다 효과적으로 세균 감염에 의한 구강질환을 예방하거나 치료할 수 있을 것이다. 특히 구강 바이오필름은 여러 세균이 모여서 바이오필름을 형성하는 특징을 가지고 있기 때문에 세포간 신호전달체계인 정족수 감지(quorum sensing)는 구강 바이오필름을 제어할 수 있는 목표점이 될 수 있다. 이외에도 구강세균의 알칼리 생성 기질인 아르기닌을 이용하여 구강 미생물의 분포를 건강한 치아와 유사한 환경으로 전환하거나 S. mutans의 glucosyltransferase 분비를 억제하여 비수용성의 글루칸 형성을 억제함으로써 바이오필름 형성을 억제하는 방법도 구강 바이오필름을 제어할 수 있는 목표점이 될 수 있다. 이처럼 구강 내 병원성 세균의 사멸을 유도하기 보다는 구강 세균의 바이오필름 형성을 억제하거나 제거하는 방법은 치아우식증을 비롯한 구강질환을 선택적으로 예방하거나 치료할 수 있는 새로운 전략이 될 것이다.