소아의 치면세균막에 존재하는 mutans streptococci의 분포

국중기,박종휘,유소영,김화숙,이난영 大韓小兒齒科學會 2004 大韓小兒齒科學會誌 Vol.31 No.3

한국인 소아의 치면세균막에 존재하는 mutans streptococci 종 및 생물형의 발현빈도와 치아우식경험지수와 상관관계를 알아보기 위하여. 조선대학교 치과병원에 내원한 12세 미만 113명의 소아 환자의 치아우식경험지수를 구하고, 이들의 상하악 유전치 및 유구치의 협면 및 설면의 치면세균막 샘플을 채취하여 mutans streptococci를 mitis-salivarius bacitracin 배지에서 선택적으로 분리하였다. 이들의 biotype을 알아보기 위해 생화학적 검사를 실하였고, 이들의 종 수준에서의 동정을 위해 dextranase 유전자를 표적으로 하는 중합효소연쇄반응을 시행하여 하였다. 113명의 환자 중에서 40명의 치면세균막에서 40 균주의 mutans streptococci이 검출되었다. 이들 중 생물형 제 Ⅰ형 (45%)이 가장 많이 검출되었으며, 그 다음으로 제Ⅳ형 (32.5%), 제 Ⅱ형 (15%). 제 Ⅴ형 (5%), 제 Ⅲ형 (2.5%) 순으로 검출되었다. 또한, 종 수준에서의 발현빈도를 알아본 결과 S. mutans가 69%, S. sobrinus는 31%였다. Mutans streptococci 종 또는 생물형에 따른 환자의 치아우식경험지수간의 차이는 없는 것으로 조사되었다(p>0.05). 이상의 결과를 종합할 때, 한국인의 소아의 구강 내에 존재하는 mutans streptococci중 생물형 제 Ⅰ형인 S. mutans가 가장 높은 빈도로 존재하며, 치아우식증이 세균학적 요소만이 아닌 기타 여러원인 요소에 의해 발병되는 다는 여러 연구 결과와 일치함을 알 수 있었다. The aim of this study is to survey the frequency of mutans streptococci species and biotypes isolated from dental plaque in Korean children and the relationship between species and biotypes of mutans streptococci and dft index. Dental plaques were collected from the anterior and molar teeth of upper and lower jaws in the subjects, aged below 12 years old. A dental examination was performed for dft (decayed, filed, total) with the WHO caries diagnostic criteria. The mutans streptococci from the sample were cultured selectively on mitis salivarius-bacitracine (MSB) agar plate. For biotyping of mutans streptococci, biochemical test was performed. From the culture, bacterial genomic DNA was prepared f3r using of PCR template for the identification of mutans streptococci at the species-level. Forty strains of mutans streptococci were isolated from dental plagues of 40 patients. The biotype Ⅰ (45%) and biotype Ⅳ (32.5%) were most frequently detected. The prevalence of s. mutans and S. sobrinus was 69% and 31%, respectively There was no positive relationship between species and biotypes of mutans streptococci and dft index. Our results revealed that biotype Ⅰ and S. mutans were frequently detected in Korean children and support that dental caries incidents by many causative factors not only bacterial factor.

Chlorhexidine과 fluoride 성분의 varnish가 타액 내 mutans streptococci 수에 미치는 효과

서정아,김재곤,백병주,양연미,이용희,김상훈 大韓小兒齒科學會 2004 大韓小兒齒科學會誌 Vol.31 No.4

본 연구의 목적은 chlorhexidine(Cervitec)과 fluoride(Fluor-protector) 성분을 각각 포함한 varnish를 치아에 적용하였을 때 타액내의 mutans streptococci의 수의 감소에 미치는 효과를 알아보기 위한 것이다. 전북대학교 치과대학생 40명을 대상으로 하여 mutans streptococci 균종 수를 평가하였으며, 이들을 무작위로 네 군으로 나누어 다음과 같이 varnish를 적용하였다; A) varnish를 적용하지 않은 군(n=10), B) chlorhexidine varnish 적용군(n=10), C) fluoride varnish 적용군(n=10), D) chlorhexidine과 fluoride varnish를 모두 적용한 군(n=10), Mutans streptococci 수 측정에는 Orion diagnostica 사의 Dentocult SM을 사용하였다. Varnish를 적용하지 않은 상태에서 1차로 Mutans streptococci 수를 측정한 후 각각의 varnish를 적용하였으며, varnish 적용 12주 후 다시 측정하여 기록하였다. 모든 실험군(group B, C, D)에서 12주 후에 측정한 Mutans streptococci 수가 감소한 개체가 증가한 개체보다 많았으나 통계적으로 유의한 차이는 보이지 않았다(p>0.05). 실험에 사용된 어떠한 varnish도 적용 12주 후에 타액 내 mutans streptococci 수에 통계적으로 유의할 만한 변화를 주지는 못하는 것으로 나타났다(p>0.05). The objective of the present prospective trial was to compare the efficacy of chlorhexidine(Cervitec), Fluoride(Fluorprotector) and combination of chlorhexdine and fluoride varnishes in decreasing the level of salivary mutans streptococci. Forty healthy students of school of dentistry, Chonbuk national university were investigated to evaluate mutans streptococci(MS) counts and randomized into four groups to treat with the experimental varnishes: A) untreated group(n=10), B) chlorhexidine group(n=10), C) fluoride group(n=10), D) chlorhexidine and fluoride group(n=10). Dentocult SM^(�)(Orion Diagnostica) strip method was used for measurement of the level of mutans streptococci in saliva. Stimulated saliva were collected at baseline for mutans streptococci counts evaluation(ms1), 12 weeks later the completion of each varnish treatment, mutans streptococci counts were re-evaluated. In varnish group with chlorhexidine, fluoride and combination of chlorhexidine and fluoride, the level of mutans streptococci was lower after 12 weeks than at baseline, but there were no significant differences in saliva(p>0.05), when compared with baseline. After 12 weeks, a remarkable reduction was still found in the subjects with high level of mutans streptococci at baseline, but not different in the low and moderate level of mutans streptococci(p>0.05).

교정환자의 브라켓과 치아 경계부에 존재하는 치면세균막내 mutans streptococci 종 및 생물형의 식별

김미애,유소영,김화숙,국중기,임성훈,윤영주,김광원 대한치과교정학회 2005 대한치과교정학회지 Vol.35 No.1

본 연구는 교정환자의 브라켓과 치아 경계부 및 브라켓으로부터 2 mm 이상 떨어진 치아 평활면의 치면세근막에 존재하는 mutaus streptococci의 종 및 생물형에 차이가 있는지를 알아보고자 시행되었다. 조선대학교 치과병원에 내원한 13세 이상 35세 미만의 환자 28명으로부터 브라켓을 장착하고 있는 61개 치아에서 치균세균막을 채취하여 mutans streptococci를 MSB 배지에서 선택적으로 분리한 다음, 이들의 지놈 DNA를 추출하여 dextranase 유전자를 표적으로 하는 중합효소연쇄반응법을 시행하고, 그 증폭물을 Hae Ⅲ로 소화하고, 이를 전기영동하여 제한효소절편길이에 따라 그 종을 식별하였다. 또한 생물형을 조사하기 위하여 생화학적 검사를 실시하였다. 그 결과 브라켓과 치아 경계부 및 브라켓으로부터 2 mm 이상 떨어진 평활면의 치면세균막에 존재하는 mutans streptococci 종은 서로 비슷한 검출 빈도를 보이나 두 곳에 존재하는 mutans streptococci 생물형은 서로 차이가 있는 것으로 나타났다. 향후 브라켓과 치아 경계부 및 치아 평활면의 치면세균막의 mutans streptococci 생물형의 차이와 브라켓 주위의 법랑질 탈회 및 치아우식증 발병과의 상관관계에 대한 연구가 필요하다. The aim of this study was to compare the species and biotypes of mutans streptococci isolated from dental plaques sampled from the interfaces between the bracket and tooth surface and smooth tooth surfaces in orthodontic patients. Dental plaque was collected from the interfaces between brackets and teeth (test group), and from smooth tooth surfaces distant from brackets by more than 2 mm (control group). The dental plaque collected by a sterilized curette was transferred into a vial of 1 X PBS. The sample in the vial was vigorously vortexed for1 min and plated on mitissalivarius bacitracin (MSB) agar plate using cotton tips. The agar plates were incubated at 37℃ in a candle jar for 2 days, and again incubated for 1 more day at anambient temperature. Individual colonies were cultured in TH broth at 37℃ CO₂ incubator. The PCR-RFLP based on dextranase gene was performed for the identification of mutans streptococci at the species-level. For biotyping of mutans streptococci, biochemical tests were performed. There was no significant difference of the species of mutans streptococci isolated from both test and control groups. However, the biotypes of the mutans streptococci isolated from test and control groups were different. These results may offer the basic data to verify the relationship between the mutans streptococci biotype and enamel decalcification or dental caries in orthodontic patients with fixed appliances.

Isolation and Characterization of the Mutans Streptococci from the Dental Plaques in Koreans

Yoo, So-Young,Park, Seon-Joo,Jeong, Dong-Ki,Kim, Kwang-Won,Lim, Sung-Hoon,Lee, Sang-Ho,Choe, Son-Jin,Chang, Young-Hyo,Park, In-Soon,Kook, Joong-Ki The Microbiological Society of Korea 2007 The journal of microbiology Vol.45 No.3

Mutans streptococci have been implicated as cariogenic bacteria in dental caries because they can produce high levels of dental caries-causing lactic acid and extracellular polysaccharide. The aim of this study was to isolate and characterize the mutans streptococci from the dental plaque obtained from Koreans. The dental plaque samples were collected from the anterior and molar teeth of both jaws in 155 subjects (aged 2 to 33.2 years, average age $13.7{\pm}4.7\;years$). The samples were diluted by 100-fold in $1{\times}\;PBS$ and plated on mitis-salivarius bacitracin (MSB) agar plates. The mutans streptococci grown on MSB plates were screened by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) targeting dextranase gene (dex). The mutans streptococci were identified at the species level using a 16S rDNA sequencing comparison method. The biochemical tests were carried out to biotype the mutans streptococci. Ninety-five strains of the mutans streptococci out of 358 colonies, which were derived from 141 subjects, were isolated. Of them, 77 strains and 18 strains were Streptococcus mutans and Streptococcus sobrinus, respectively. The biotyping data showed that 62, 1, 20, 10, and 2 strains were biotypes I, II, IV, V and variant, respectively. Of the two strains of variant biotype, one strains was similar to biotype IV except that it was positive to the arginine hydrolysis test. We considered this one strain a new biotype, and classified it as biotype VII. In conclusion, S. mutans and its biotype I was most frequently isolated in Korean dental plaque. The mutans streptococci strains isolated in this study might be useful for the study of the pathogenesis and the prevention of dental caries.

Isolation and Characterization of the Mutans Streptococci from the Dental Plaques in Koreans

유소영,Seon Joo Park,Dong Ki Jeong,김광원,임성훈,이상호,Son-Jin Choe,장영효,박인순,국중기 한국미생물학회 2007 The journal of microbiology Vol.45 No.3

Mutans streptococci have been implicated as cariogenic bacteria in dental caries because they can produce high levels of dental caries-causing lactic acid and extracellular polysaccharide. The aim of this study was to isolate and characterize the mutans streptococci from the dental plaque obtained from Koreans. The dental plaque samples were collected from the anterior and molar teeth of both jaws in 155 subjects (aged 2 to 33.2 years, average age 13.7±4.7 years). The samples were diluted by 100-fold in 1x PBS and plated on mitis-salivarius bacitracin (MSB) agar plates. The mutans streptococci grown on MSB plates were screened by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) targeting dextranase gene (dex). The mutans streptococci were identified at the species level using a 16S rDNA sequencing comparison method. The biochemical tests were carried out to biotype the mutans streptococci. Ninety-five strains of the mutans streptococci out of 358 colonies, which were derived from 141 subjects, were isolated. Of them, 77 strains and 18 strains were Streptococcus mutans and Streptococcus sobrinus, respectively. The biotyping data showed that 62, 1, 20, 10, and 2 strains were biotypes I, II, IV, V and variant, respectively. Of the two strains of variant biotype, one strains was similar to biotype IV except that it was positive to the arginine hydrolysis test. We considered this one strain a new biotype, and classified it as biotype VII. In conclusion, S. mutans and its biotype I was most frequently isolated in Korean dental plaque. The mutans streptococci strains isolated in this study might be useful for the study of the pathogenesis and the prevention of dental caries.

녹차 및 결명자 추출물의 교정용 브라켓과 치면 사이의 경계부에서 분리된 mutans streptococci에 대한 항균작용

임성훈,서정순,윤영주,김광원,유소영,김화숙,국중기,이병래,차종희,박재윤 대한치과교정학회 2003 대한치과교정학회지 Vol.33 No.5

본 연구는 치아우식증에 관련된 Mutans streptococci 표준균주 및 임상분리 균주의 성장억제를 유도할 수 있는 천연 생약추출물을 검색하기 위하여 실시되었다. 녹차추출물은 CHMC-2032를 사용하였으며, 결명자 추출물은 50% 에탄올을 이용하여 얻었다. 이들 추출물의 Mutans streptococci 표준균주 및 교정환자들에서 브라켓과 치면 사이의 경계부에서 분리된 각각 10 균주씩의 S. mutans 및 S. sobrinus에 대한 최소성장억제농도를 액체배지 희석법으로 구하였다. 그 결과 CHMC-2032의 S. mutans 및 S. sobrinus의 표준 균주, S. sobrinus의 대부분 임상분리 균주에 대한 최소성장억제농도는 5 mg/ml이었다. 그러나 결명자 추출물에 의한 S. mutans 및 S. sobrinus에 대한 세균 성장억제 효과는 미미하였다. 본 연구 결과 치아우식증의 예방적 측면에서 결명자차보다는 녹차를 마시는 것이 유리하며, CHMC-2032를 이용하여 구강양치용액을 제조하여 사용할 경우 교정환자를 포함하여 대부분의 사람에게서 치아우식증 예방 효과가 있으리고 추정된다. Mutans streptococci is the major causative factor in dental caries. Especially, orthodontic patients with fixed appliance are a risk group for dental caries. Because fixed appliances attached on teeth may change the environment of dental plaque, the enamel decalcification or dental caries around the bracket and band is a major side effect of orthodontic treatment. The aim of this study was to search plant extracts that have antimicrobial effect on mutans streptococci. Seed-extract of Casio tora were prepared with ethanol and CHMC-2032, the leaf-extracts from Camellia sinensis extract, was obtained extract, 2 type strains and 20 clinical isolates of mutans streptococci isolated from the interface between orthodontic brackets and tooth surfaces in the orthodontic patients were used in this study. The minimal inhibitory concentration of CHMC-2032 was 5 mg/ml on the S. mutans KCTC 3065, S. sobrinus KCTC 3088, and 8 clinical isolates of S. sobrinus. However, there was no antibacterial effect of seed-extract of C. tora on mutans streptococci. These data suggest that green tea may be more effective than the tea prepared from C. tora in the prevention of enamel decalcification or dental caries around brackets.

불화규산의 mutans streptococci에 대한 항균작용 및 인체돌연변이 유발가능성에 관한 연구

박은아,박근영,박병래,김유영,윤현경,최기운,이진용 대한구강보건학회 2001 大韓口腔保健學會誌 Vol.25 No.4

Antibacterial effect of F^- at lower concentrations present in fluoridated water and local application of F^- on cariogenic bacteria is controversial. The present study was performed to examine antibacterial effect of fluorosilicic acid at various concentrations and long-term effect of the fluoride at lower concentrations on representative strains of mutans streptococci: S. mutans NCTC 10449 and S. sobrinus 6715. For the purpose, physico-chemical characteristics of the bacteria treated with fluorosilicic acid were observed. Minimum inhibitory concentration (MIC) of F^- for S. mutans and S. sobrinus was determined to be 50 and 100 ppm, respectively, as measured by optical density. The addition of F^- at MIC to mutans streptococci in the exponential phase decreased the cell viability by more than 80%. F^- in combination with xylitol and polyphosphate slightly enhanced their antibacterial activity. The experimental strains exposed to F^- (0∼15 ppm) for 2 years on blood agar plates containing fourosilicic acid demonstrated to obtain resistance to some extent to F^-. F^- exposure decreased the inhibition of the bacterial growth by F^- as observed after the incubation with F^- at the range of MIC, the concentration determined 2 years ago. The decrease in the inhibition of growth and survival by F^- was more prominent with the strains long-term exposed to higher concentrations of F^-. The pH of the F^- -exposed strains when cultured in broth containing fluorosilicic acid was significantly lower than that of the strains which had not been exposed to F^-. Difference in pH between 4- and 24-h incubation was more prominent with the strains long-term exposed to F^-, suggest that F^- exposure conferred F^- resistance to the mutans streptococci. The higher concentrations of F^- the strains exposed to, the higher synthesis of soluble glucan was measured in S. mutans, resulting in higher binding of the bacterium to glass surface. However, those were decreased in S. sobrinus long-term exposed to higher concentrations of F^-. Regardless whether the bacteria were exposed to F^- or not, no significant difference in binding to saliva-coated hydroxyapatite and protein profiles. Transmission electron micrographs revealed that loss of the cell wall and vacuolization in the cytoplasm were distinct in the bacteria exposed to higher concentrations of F^-. It was shown in Ames test that F^- up to 100 ppm appeared to be negative for mutagenicity. Overall, the results suggest that combination of fluorosilicic acid with other anticariogenic reagents may be benefical for a better caries prevention. Use of fluoride for long-term period of time even at lower concentrations may contribute to the efficacy of fluoride in caries prevention, inhibiting the bacterial function and metabolism.

시각적 염색 방법을 이용한 마우스워시의 구강균에 대한 항균효과 확인

박태훈 ( Taehun Park ),조정훈 ( Jeong Hun Cho ),성영은 ( Youngeun Sung ),조준철 ( Jun-cheol Cho ),신계호 ( Kyeho Shin ) 대한화장품학회 2014 대한화장품학회지 Vol.40 No.2

충치는 사람의 구강질환 중 가장 흔한 질환으로 Streptococcus mutans (S. mutans)균이 초기 충치를 형성하는데 매우 중요한 역할을 담당한다. Porphyromonas gingivalis (P. gingivalis)는 대표적인 구취 유발균으로 구취 형성에 중요한 휘발성 황화합물을 생성하는데 관여한다. 치주질환은 치은결체조직과 치조골의 파괴를 유발하여 치아의 상실을 초래할 수 있는 만성 염증성 질환으로 Prevotella intermedia (P. intermedia)가 원인균이다. 이번 연구에서는 cetylpyridinium chloride (CPC), sodium fluoride (NaF), 녹차 추출액, 솔잎 추출액을 유효성분으로 하는 마우스워시 제품을 사용하여 S. mutans 균을 포함, 구강질환 균으로 널리 알려진 P. gingivalis, P. intermedia 대해 항균 효과를 확인하고자 하였다. 그 결과 시험군의 경우 S. mutans, P. gingivalis에 대해 30 s 내에 4.00 Log, 4.68 Log의 사멸력을 확인하였고, P. intermedia의 경우 30 s 2.40 Log, 60 s 2.70 Log 사멸력을 확인하였다. 또한 Dentocult SM Strip mutans (SM Strip) 염색방법을 적용하여 S. mutans 균의 감소여부를 시각적 자료로 쉽게 확인할 수 있었다. 이와 같은 결과를 통해 CPC, NaF, 녹차 추출액, 솔잎 추출액을 포함한 마우스워시 제품은 구강균 사멸을 통해 충치 및 구취와 같은 구강질환 예방에 효과가 있을 것으로 기대한다. Dental caries are one of the most common oral diseases and Streptococcus mutans (S. mutans) plays an important role in the initiation and progression of dental caries. Oral malodor is primarily the result of microbial metabolism such as Porphyromonas gingivalis (P. gingivalis) that produce volatile sulfur compounds (VSCs), causing oral malodor. Prevotella intermedia (P. intermedia) is known as typical periodontopathic bacteria, and periodontal disease is chronic inflammatory disease that leads to damage of gingival connective tissue and alveolar bone, eventually loss of teeth. In this study, we investigated antimicrobial effect of mouthwash containing cetylpyridinium chloride (CPC), sodium fluoride (NaF), green tea water extract and pine needles water extract against cariogenic and periodontopathic bacteria sucn as S. mutans, P. gingivalis and P. intermedia. As a result, the reduction ratios of S. mutans and P. gingivalis were 4.00 Log and 4.68 Log reduction for 30 s, and P. intermedia were 2.40 Log reduction for 30 s and 2.70 Log reduction for 60 s. Dentocult SM Strip mutans (SM Strip) provides easy detection of visual data showing a significant inhibition on S. mutans. In conclusion, we expected that mouthwash containing CPC, NaF, green tea water extract and pine needles water extract could help preventing the dental disease like dental caries and oral malodor.

혼합치열기 어린이에서 mutans streptococci에 대한 자일리톨 껌의 향균효과에 관한 연구

박재홍 大韓小兒齒科學會 2007 大韓小兒齒科學會誌 Vol.34 No.4

Xylitol 껌의 mutans streptococci(MS)에 대한 항균효과를 평가하기 위하여 6세∼11세 사이의 혼합치열기 어린이 62명을 선정하였다. 4개월 동안 1개의 xylitol 껌을 하루 3회 씹는군,2개의 xylitol 껌을 하루 3회 씹는 군과 xylitol 껌을 씹지 않는 대조군으로 나누어 하루 3번 식후 최소 5분간 껌을 씹게 한 후 1개월 간격으로 자극성타액을 채취하여 선택배지 (mitis salivarius kanamicin bacitracin agar)에 배양한 후 MS 군락 수의 감소를 평가하였다. 1회 1개의 xylitol 껌을 씹는 군에서는 MS의 감소를 보이지 않았고,1회 2개의 xylitol 껌을 씹는 군에서 4개윌째 MS의 유의한 감소를 보였다(P < 0.05). 따라서 혼합치열기 어린이의 경우 하루 3회,총 10g 정도의 xylitol을 4개월 이상 꾸준히 사용하였을 경우 치아우식증에 대한 예방효과가 있을 것으로 사료된다. The purpose of this study was to investigate the effect of xylitol chewing gums on mutans streptococci(MS) counts in saliva. Sixty two children 6 to 11 years old were randomly assigned into one of three groups, Stimulated saliva specimens were plated in duplicate on conventional selective culture (mitis salivarius kanamycin bacitracin agar) for mutans streptococci. Polymerase chain reaction (PCR) amplication was performed to identify MS. After the 4-month period a significant decrease of the MS counts occurred in the group B (two gum 3 times a day: P < 0.05) but not in group A (one gum 3 times a day) and control group (didn t receive xylitol gum). According to qualitative evaluations, xylitol use did reduce the levels of MS in mixed dentition children. It has been suggested that a daily intake of 2 tablet for 3 times a day (about lOg) is needed in order to obtain a clinical anticariogenic effect.

Whole-genomic DNA probe를 이용한 구강 Streptococcus 균종의 식별

이영호,이장희,김각균,최선진 대한구강생물학회 1992 International Journal of Oral Biology Vol.16 No.1

DNA hybridization with whole-genomic DNA probe and restriction pattern analysis were done in search for a useful method for identification of the species of mutans group streptocci. Dot blots of genomic DNA of five strains (serotypes a, b, c and g) of mutans streptococci and on strain of Streptococcus sanguis were hybridized with ^32P-labeled whole-genomic DNA of Streptococcus mutans strain 10449 (serotype c) as a probe. Also, genomic DNAs of each strain were digested with two restriction enzymes (EcoRI and HindIII) and the restriction patterns were compared on 0.7% agarose gel after electrophoresis. Whole-genomic DNA probe of Strep. mutans strain 10449 was able to detect 3ng of homologous DNA per dot and was also able to detect the same amount of genomic DNA of a different strain of the same serotype with about the same intensity as shown on the autoradiogram. Genomic DNAs of differnet serotypes of mutans streptococci and of Strep. sanguis strain MPC1 were hybridized weakly with the probe at 50-100ng per dot. Genomic DNAs of six bacteria of two differnet genera (Bacteroides gingivalis and intermedius) were not hybridized with the probe at any concentration used in experiment. Restriction-digested patterns of genomic DNAs were all different from strain to strain, indicating genomic structures between each serotypes of mutans streptococci.