• 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.

• 치위생과학회지
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• 제15권2호
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• pp.232-237
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• 2015

이 연구는 강릉원주대학교 치과대학 학생들의 임상실습을 위해 사용되고 있는 DCU에서 배출되는 물 속 종속영양세균의 수준을 평가하면서 사용빈도에 따른 세균 오염수준의 차이를 확인하고 기회 감염성 병원균의 존재를 분자생물학적 방법을 사용하여 확인하였다. 임상 실습실에서 사용되는 DCU 36개를 대상으로 초음파치석제거기에서 물 시료를 수집하여 평균 CFU/ml를 조사하고 초음파치석제거기의 한달 사용빈도에 따라 DCU를 세 집단으로 분류하여 세균오염수준을 비교하였다. 또한 수집한 물 시료에서 세균의 genomic DNA를 추출한 후 PCR 분석을 통해 기회감염성 병원균의 존재를 확인한 결과는 다음과 같다. 학생 실습에 사용한 DCU에서 수집한 물 시료의 평균 종속영양세균수준은 16,095 CFU/ml로 ADA에서 권장하는 200 CFU/ml 이하의 수준에 적합하지 않은 것을 확인하였다. 초음파 치석제거기의 한 달 사용빈도에 따라 3집단으로 나누어 CFU/ml를 조사하였을 때, 초음파치석제거기를 한 달에 1번 이상 3번 미만 사용한 DCU에서 평균 CFU/ml가 20,070 CFU/ml로 가장 높게 나타났으며, 3번 이상 사용한 유니트는 CFU/ml 평균이 8,420 CFU/ml로 가장 적게 나타났다. 3개군의 CFU/ml 차이는 통계학적으로 유의성이 있는 것을 보여주었고(p<0.05), 그 중 사용빈도가 가장 높은 군에서 유의하게 낮은 CFU/ml를 보여주었다. 치과에서 사용하는 DCU에 존재하는 기회감염성 병원균이 학생실습에 사용하는 DCU에서도 분리되었다. 36개의 genomic DNA 시료 중 1개의 시료에서 Pseudomonas species가 검출되었고, 2개의 시료에서 비결핵성 Mycobacterium species가 검출되었다. 따라서 학생실습용으로 사용되는 DCU는 학생들과 대상자에게 잠재적 감염의 원인이 될 수 있으며, 실습 전 학생들의 보호장비 착용과 실습 후 수관관리가 필요하다.

• International Journal of Oral Biology
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• 제37권3호
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• pp.103-108
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• 2012

The purpose of this study was to assess the efficacy of photodynamic therapy (PDT) using erythrosine and a halogen light source to treat a biofilm formed on a machined surface titanium disk in vivo. Ten volunteers carried an acrylic appliance containing six machined surface titanium disks on the upper jaw over a period of five days. After the five days of biofilm formation period, the disks were removed. PDT using 20 ${\mu}M$ erythrosine and halogen light was then applied to the biofilms formed on the disks. Experimental samples were divided into a negative control group (no erythrosine and no irradiation), E0 group (erythrosine 60s + no irradiation), E30 group (erythrosine 60s + halogen light 30s), and E60 group (erythrosine 60s + halogen light 60s). Following PDT, the bacteria in the biofilm were found to be detached from each disk. Each suspension with detached bacteria were diluted and cultivated on a blood-agar plate for five days under anaerobic conditions. The cultivated bacterial counts in the E60 group were significantly lower than the control group (86.4%) or E0 group (76.7%). In the experimental groups also, the light exposure time and bacterial counts showed a negative correlation. In conclusion, PDT using erythrosine and halogen light has bactericidal effects on biofilms formed on a titanium disk in vivo. Notably, applying 20 ${\mu}M$ erythrosine and 60 seconds of halogen light irradiation had a significantly potent effect.

• Journal of Dental Anesthesia and Pain Medicine
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• 제16권1호
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• pp.17-24
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• 2016

Background: To evaluate the antimicrobial activity of lidocaine (LD) topical anesthetic spray against oral microflora. Methods: Antimicrobial effects of 10% LD spray were assessed against six bacterial cultures obtained from volunteers: Escherichia coli, Enterococcus faecalis, Staphylococcus aureus, Streptococcus salivarius, Streptococcus pyogenes, and Streptococcus sanguinis. The filter papers contained $50-{\mu}l$ LD, brain heart infusion (BHI) broth, or 0.2% chlorhexidine. Papers were placed on the cultured blood plates for 1-3 min. After the papers were removed, plates were incubated for 24 h. Bacterial growth on the contact areas was recorded as the antimicrobial score. The split mouth technique was use in for sample collection in clinical study. Filter papers soaked with either BHI broth or LD were placed on the right or left buccal mucosa for 1 min, and replaced with other papers to imprint biofilms onto the contact areas. Papers were placed on blood plates, incubated for 24 h, and antimicrobial scores were determined. Experiments were conducted for 2- and 3-min exposure times with a 1-day washout period. Results: LD exhibited bactericidal effects against E. coli, S. sanguinis, and S. salivarius within 1 min but displayed no effect against S. aureus, E. faecalis, and S. pyogenes. The antimicrobial effect of LD on oral microflora depended upon exposure time, similar to the results obtained from the clinical study (P < 0.05). LD showed 60-95% biofilm reduction on buccal mucosa. Conclusions: Antimicrobial activity of 10% LD topical anesthetic spray was increased by exposure time. The 3 min application reduced oral microflora in the buccal mucosa.

• Asian Pacific Journal of Cancer Prevention
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• 제17권9호
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• pp.4491-4501
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• 2016

Verruco-papillary lesions (VPLs) of the oral cavity described in the literature involve a spectrum of conditions including squamous papilloma, verruca vulgaris, focal epithelial hyperplasia, condyloma, proliferative verrucous leukoplakia and verrucous carcinoma. A majority of the VPLs are slow growing, benign in nature and have a viral aetiology. Virus associated benign mucosal outgrowths are not too difficult to diagnose either clinically or by microscopy. Apart from virus-associated lesions, VPLs harboring malignant potential or behaviour such as verrucous carcinoma, proliferative verrucous leukoplakia, oral verrucous hyperplasia (OVH), oral papillary squamous cell carcinoma (PSCC) and oral conventional squamous cell carcinoma with papillary features (CSCC) need to be further clarified for better understanding of their predictable biologic behavior and appropriate treatment. Current understanding of potentially malignant VPLs is perplexing and is primarily attributed to the use of confusing and unsatisfactory terminology. In particular, the condition referred to as oral verrucous hyperplasia (OVH) poses a major diagnostic challenge. OVH represents a histopathological entity whose clinical features are not well recognised and is usually clinically indistinguishable from a verrucous carcinoma and a PSCC or a CSCC. A consensus report published by an expert working group from South Asia as an outcome of the 'First Asian Regional Meeting on the Terminology and Criteria for Verruco-papillary Lesions of the Oral Cavity' held in Kuala Lumpur, Malaysia, recognised the clinical description of these OVH as a new entity named 'Exophytic Verrucous Hyperplasia'. Previously described clinical features of OVH such as the 'blunt' or 'sharp' variants; and the 'mass' or 'plaque' variants can now collectively fall under this newly described entity. This paper discusses in detail the application of the standardized criteria guidelines of 'Exophytic Verrucous Hyperplasia' as published by the expert group which will enable clinicians and pathologists to uniformly interpret their pool of OVH cases and facilitate a better understanding of OVH malignant potential.

• Advances in Traditional Medicine
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• 제4권3호
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• pp.179-185
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• 2004

The ethanol extract of Indigofera cordifolia was studied for in vivo gastroprotective activity, cytotoxic activity against oral tumor and normal cells, multidrug resistance (MDR) reversal activity, anti-human immunodeficiency virus (HIV) activity and radical scavenging activity. The extract of I. cordifolia showed potent gastric mucosal protective activity against stomach injury induced by HCl/EtOH solution. However, the gastroprotective activity could not be related to the radical mechanism, because the extract weakly scavenged both OH radical and $O_2*^-$. The extract also showed promising levels of MDR-reversing activity. This study demonstrates the tumor-specific cytotoxic action of the plant extract. However, the extract had no anti-HIV activity. From above results, the study suggests the medicinal importance of I. cordiforia extract.

• 대한미생물학회지
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• 제21권1호
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• pp.63-71
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• 1986

The antifungal activities of amphotericin B, 5-fluorocytosine, and ketoconazole in combination of amphotericin B/ketoconazole and 5-fluorocytosine/ketoconazole were determined against 42 strains of Candida spp. isolated from oral cavity. Among 42 strains of Candida species, 36 strains of Candida albicans, 2 strains of Candida parapsilosis and Candida tropicalis 1 strain of Candida krusei and Candida stellatoidea were identified. The minimum inhibitory concentrations(MICs) of amphotericin B, 5-fluorocytosine and ketoconazole for these strains were ranged from 0.05-1.56 mcg/ml, 12.5->100.0 mcg/ml and 0.2-50.0 mcg/ml. In all of the experimental strains, amphotericin B had the greatest antifungal activity on a dilution basis. When a microtiter checkerboard technique was used 5-fluorocytosine acted synergistically with ketoconazole against all strains, whereas amphotericin B has a reduced effect. The killing curve experiments with on strain of Candida albicans WMC-85024 demonstrated that the combination of amphotericin B/ketoconazole and 5-fluorocytosine/ketoconazole produced a decrease in number of colony forming unit of >3 logs in 72 hours.

• 대한미생물학회지
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• 제22권4호
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• pp.403-411
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• 1987

This study investigated whether a correlation exists between proteinase activity, phospholipase activity and adherence of Candida albicans to buccal epithelial cells by using of various strains isolated from oral cavity. The proteinase activity of 30 strains was tested by culture on agar media that contained bovine serum albumin as a nitrogen source. Using the serum-protein-agar method to test proterolysis of serum albumin in 20 strains of Candida albicans. Twenty-six strains of Candida albicans were phospholipase producers and the degree of phopholipase activity of experimental strains were $0.51{\sim}0.89$ measured by Pz-value. Twenty-eight strains of Candida albicans were adhersive to buccal epithelial cells and 15 strains were foung significantly active adherence. Fifteen strains of Candida albicans were correlated with proteinase activity and adherence to epithelial cells and concomitantly 20 strains of Candida albicans were also correlated with phospholipase activity and adherence. In conclusion our investigation provides evidence of a correlation between quantitative proteinase, phospholipase and adherence. An association of these parameters may be an important contributory factor for pathogenicity.

• 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.

• Asian Pacific Journal of Cancer Prevention
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• 제17권7호
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• pp.3335-3340
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• 2016

Background: Oral submucous fibrosis (OSF) is a precancerous condition with a 4 to13% malignant transformation rate. Related to the habit of areca nut chewing it is mainly prevalent in South-east Asian countries where the habit of betel quid chewing is frequently practised. On chewing, alkaloids and polyphenols are released which undergo nitrosation and give rise to N-nitrosamines which are cytotoxic agents. CYP450 is a microsomal enzyme group which metabolizes various endogenous and exogenous chemicals including those released by areca nut chewing. CYP1A1 plays a central role in metabolic activation of these xenobiotics, whereas CYP2E1 metabolizes nitrosamines and tannins. Polymorphisms in genes that code for these enzymes may alter their expression or function and may therefore affect an individuals susceptibility regarding OSF and oral cancer. The present study was therefore undertaken to investigate the association of polymorphisms in CYP1A1 m2 and CYP2E1 (RsaI/PstI) sites with risk of OSF among areca nut chewers in the Northern India population. A total of 95 histopathologically confirmed cases of OSF with history of areca nut chewing not less than 1 year and 80, age and sex matched controls without any clinical signs and symptoms of OSF with areca nut chewing habit not less than 1 year were enrolled. DNA was extracted from peripheral blood samples and polymorphisms were analyzed by PCR-RFLP method. Gene polymorphism of CYP1A1 at NcoI site was observed to be significantly higher (p = 0.016) in cases of OSF when compared to controls. Association of CYP1A1 gene polymorphism at NcoI site and the risk of OSF (Odd's Ratio = 2.275) was also observed to be significant. However, no such association was observed for the CYP2E1 gene polymorphism (Odd's Ratio = 0.815). Our results suggest that the CYP1A1 gene polymorphism at the NcoI site confers an increased risk for OSF.