• Restorative Dentistry and Endodontics
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• 제21권2호
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• pp.470-488
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• 1996

The end products of the metabolism of the oral microorganism, organic acids, are an element that produces dental caries. Four organic acids in plaque fluid, lactic acid, acetic acid, succinic acid, propionic acid which take the important role in producing dental caries, were chosen to evaluate the effect of acid type and concentration. The subject, $100{\mu}m$ in thickness, were immersed in acid-buffer solution which has the different acid concentration of 10mM, 25mM, 50mM, 100mM and pH 4.3 and degree of saturation was $0.153{\pm}0.003$ kept in constant and were operated to produce artificial caries under different demineralization time (1, 2, 3 day) at x25. The results were obtained by observing under polarizing microscope at x25. 1. The subsurface lesion, specific finding of incipient enamel caries, showed positive birefringence. but surface zone and sound enamel showed negative birefringence. 2. The demineralization rate of enamel was increased as the acid concentration increased. 3. The subsurface lesion showed increasing depth in the order of lactic, acetic, propionic acid, succinic acid. 4. The concentration of organic acid in artificial caries system had an independent effect on demineralization rate in enamel under the constant pH and degree of saturation. The result of this study showed that not only pH and the acid strength but the concentration of organic acid had an independent effect on demineralization rate in early enamel caries. And through the further research on the factors influencing enamel demineralization, it will be necessary to develop an effective caries preventive therapy.

• 한국치위생학회지
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• 제5권2호
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• pp.131-145
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• 2005

There are normal inhabitants doing medically useful functions in the body. There are many kinds of bacteria performing specific functions in the oral cavity. Two strains of lactic add bacteria were isolated from normal inhabitants of children's oral cavity, which inhibited the production of volatile sulfur compounds by anaerobic bacteria. The authors identified the isolates by 16S rDNA partial sequencing. 1. Two isolates were Gram-positive bacilli and produced hydrogen peroxide. 2. When Streptococcus mutans was cultured in the media, the mean weight of formed artificial plaque on the orthodontic wires was $124.4{\pm}30.4$ mg, whereas being reduced to $5.2{\pm}2.0$ mg and $10.6{\pm}6.6$ mg in the media cultured with Streptococcus mutans and each isolate, respectively(p<0.05). 3. The number of viable cells of Streptococcus mutans was $3.4{\times}10^9$ per ml in the cultured solution, whereas those of Streptococcus mutans in the combined culture with each of isolates were $4.6{\times}10^8$ and $2.4{\times}10^8$ per ml. 4. The optical density was 1.286 in the supernatant of Fusoacterium nucleatum after vortexing for 30 minutes, whereas in the supernatant of combined Fusoacterium nucleatum and each isolate, they were reduced to 0.628 and 00497, which the percentages of coaggregation between them were 2904% and 57.8%, respectively. 5. The optical density of Fusoacterium nucleatum precipitate was 1.794 in the culture media containing cysteine and $FeSO_4$ being reduced to 1.144 and 0.915 in the coaggregated precipitates of Fusoacterium nucleatum and each isolate. 6. The similarity values of 16S rDNA sequence between each of isolates and Lactobacillus salivarius subsp. salicinius were 99.60% and 99.73%, respectively, meaning that isolates were Lactobacillus salivarius subsp. salicinius. These results indicated that two strains isolated from children's saliva, which inhibited the formation of plaque and the production of volatile sulfur compounds, were identified as Lactobacillus salivarius subsp. salicinius.

• 대한소아치과학회지
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• 제27권1호
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• pp.15-23
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• 2000

경조직 우식으로 발생하는 치아우식증은 다인성 질환으로서 치태내 세균중에서도 Streptococcus mutans가 주 원인균이며, 치면에 부착, 증식 및 산생성 과정을 거쳐 치아우식을 유발한다. Streptococcus salivarius는 사람의 구강에 정상적으로 존재하는 세균이다. 본 연구에서는 소아의 구강으로부터 분리된 Streptococcus salivarius 119의 특성과 Streptococcus mutans 및 Streptococcus oralis에 대한 영향을 연구하여 다음과 같은 결과를 얻었다. 1. Streptococcus matans를 일회용 큐벧에서 배양시 550nm에서의 흡광도가 0.327이었으나, Streptococcus mutans와 Streptococcus salivarius 119의 혼합 배양시에는 0.119로 감소되었다. 2. 비커 와이어 검사에서 Streptococcus mutans 배양시 형성된 인공치태 무게는 84.7mg이었으나, Streptococcus mutans와 Streptococcus salivarius 119 혼합 배양시에는 12.3mg으로 감소되었다. 3. BHI broth에서 배양된 Streptococcus salivarius 119 배양 상청액을 가한 비커 와이어 검사에서 형성된 인공치태 무게는 100.5mg인데 반해, 5% 자당이 함유된 BHI broth에서 배양된 Streptococcus salivarius 119 배양 상청액을 가한 비커 와이어 검사에서 20.4 mg이었다. 4. Streptococcus oralis와 Streptococcus salivarius 119 단독 배양시에는 각각 ml당 $4.8\times10^7,\;7.5\times10^8$이었으나, 혼합 배양시에는 Streptococcus oralis는 $4.2\times10^7$, Streptococcus salivarius 119는 $5.8\times10^7$로 감소하였다. 5. Streptococcus salivarius 119 배양 상청액을 thin layer chromatography를 실시한 결과, Streptococcus salivarius 119가 형성한 polymer는 글루캔이었다. 6. Streptococcus salivarius 119가 만드는 글루캔을 처리하여 thin layer chromatography를 실시한 결과, $1\rightarrow6$ 결합이 주된 결합인 수용성 글루캔이었다. 이상의 결과를 종합하면 구강에서 분리된 Streptococcus salivarius 119에 의한 Streptococcus mutans의 인공치태 형성 억제작용은 수용성 글루캔 형성에 의한 것으로 사료되었다.

• 대한치과보철학회지
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• 제44권1호
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• pp.124-134
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• 2006

Statement of problem: Streptococcus produces energy and forms extracellular polysaccharides by metabolizing sucrose. Insoluble glucan, a kind of extracellular polysaccharide, is the important material of dental plaque. Fructose affects the metabolism of sucrose. Purpose: The purpose of this study was to evaluate the effect of fructose on the metabolism of sucrose in Streptococcus mutans. Materials and methods: To determine the effect of fructose on the formation of artificial plaque by Streptococcus mutans Ingbritt, S. mutans and fructose were placed in beakers containing M17 broth and sucrose. The wires were hung on frameworks inserted into cork stoppers, and then immersed in each of the beakers. After the incubation with gentle shaking, each wire was weighed. To analyze the effect of fructose on the sucrose metabolism by S. mutans or glucosyltransferase, S. mutans and fructose were placed in M17 broth containing sucrose. After the incubation. the remaining sucrose and polymers were analysed by thin layer chromatography. Results: The following results were obtained; 1. When Streptococcus mutans was cultured in the media containing 3% sucrose for 8 hours, the mean weight of formed artificial plaque on the wires was $124.3{\pm}3.0mg$, whereas being reduced to $20.7{\pm}10.2mg$ in the media added with 3% sucrose and 4% fructose(p<0.05) 2. When the control containing glucose was added with sucrose, the optical density of Streptococcus mutans solution cultured for 24 hours was not increased compared with the control, while being increased by adding with fructose. 3. When Streptococcus mutans was incubated in the media added with sucrose and fructose for 8 hours, the number of viable cells was increased compared with the media added with sucrose. 4. The amount of remained sucrose was increased in Streptococcus mutans culture supernatant of media added with sucrose and fructose than with sucrose only. but the amount of produced insoluble glucan was decreased. 5. The amounts of remained sucrose and produced soluble glucan were increased in the culture of glucosyltransferase-contained media added with sucrose and fructose than with sucrose only, but the amount of produced insoluble glucan was decreased . Conclusion: These results indicated that the sucrose metabolism and the production of insoluble glucan were inhibited in Streptococcus mutans by adding fructose in the media containing sucrose.

• Journal of Periodontal and Implant Science
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• 제30권2호
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• pp.403-420
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• 2000

The treatment and prevention of periodontitis is focused on the reduction and the elimination of pathogenic bacteria, especially A. actinomycetemcomitans and black pigmented bacteria such as P. gingivalis. To prevent recurrent disease, the recolonization of these bacteria should be inhibited in the periodontal pocket. Since the replacement therapy was introduced in periodontics by Hillman et al, Jeong et al reported that hydrogen peroxide-producing Lactobacillus acidophilus V-20 completely inhibited P. gingivalis and A. actino - mycetemcomitans in vitro and mouth gargling with Lactobacillus acidophilus V-20 in periodontitis patients during the maintenance phase improved clinical condition and reduced the No. of P. gingivalis and A. actinomycetemcomitans at 4 weeks of treatment. Prior to replacement therapy with bacteria, dynamics of microbial colonization should be considered. This study was performed to evaluate the change in the viable cell number of Lactobacilli and P. gingivalis after oral administration of L. acidophilus V-20. In periodontal health, gargling increased the No. of Lactobacilli in saliva, buccal mucosa, supragingival plaque from the first week, which maintained for 2-3 weeks after gargling stop, and then returned to the undetectable baseline level at the ninth week. In the periodontal pocket of moderate periodontitis patients, daily irrigation for 1 week and weekly irrigation for subsequent 3 weeks decreased the viable cell number of P. gingivalis during the period of irrigation and increased the number of Lactobacilli, which was maintained from the second to the seventh week. L. acidophilus V-20 was isolated for the first 2 weeks of oral administration, and the 3 different strains of Lactobacilli were isolated continuously for remaining period and identified as L. ali - mentarius, L. casei subspecies casei and L. fructosus. The first two Lactobacilli strains completely inhibited P. gingivalis in vitro and all the isolated Lactobacilli strains reduced the artificial plaque formation by 55-63%. These results showed that mouth gargling or pocket irrigation with L. acidophilus V-20 increased the No. of intraoral Lactobacilli and caused to decrease in the No. of P. gingivalis. This suggests that the replacement therapy by these Lactobacilli might be useful in the maintenance care of periodontal patients.

• 한국미생물·생명공학회지
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• 제24권1호
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• pp.9-18
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• 1996

Streptococcus mutans has been implicated as primary causative agents of dental caries by insoluble glucan (IG) in human and experimental animals. An attempt was made to search for the ${\alpha}$-1,3 glucanase that degrades IG produced by S. mutans. ${\alpha}$-1,3 glucanase was detected in the culture supernatant of microorganisms, which are isolated from soils on agar medium containing IG as a sole carbon source. This Streptomyces sp. hydrolysed IG produced by immobilized S. mutans and was named as Y9373. This enzyme required ${\alpha}$-1,3 glucan (IG) as an inducer. The optimum conditions for enzyme production were studied. The enzyme was purified by 30~70% $(NH_4)_2SO_4$ precipitation, anion exchange chroma tography on DEAE-cellulose and gel filtration on Sepadex G-75. The purified enzyme has a specific activity of 7840.0 U/mg protein giving 32.1-fold purification and final yield of 0.53%. The molecular weight was estimated to be about 22.5 kDa by SDS-PAGE. The optimum pH and temperature for enzyme reaction were 6.5 and 37$^{\circ}C$, respectively and the enzyme was relatively stable at the temperature below 60$^{\circ}C$. The activity of purified enzyme was enhanced by adding $Co^{2+},\;Mn^{2+}\;and\;Mg^{2+}$ into the medium, whereas inhibited by adding $Hg^{2+},\;Zn^{2+}$ and SDS. The $K_m\;and\;V_{max}$ value of ${\alpha}$-1,3 glucanase for IG were estimated to be 2.50 mM and 0.0431 mM/min, respectively. The thin layer chromatographic analysis of hydrolysates from IG with ${\alpha}$-1,3 glucanase showed that glucose was the main product of reaction. This enzyme activity was about 14 times higher than marketing dextranase as preventive agent against artificial dental caries by S. mutans in TH medium including 5% sucrose after 30 minutes.

• Journal of Periodontal and Implant Science
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• 제27권1호
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• pp.61-78
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• 1997

PDGF-BB has been recognized as a highly potential growth factor for guided tissue regeneration in periodontal defect. This study carried out histologic and histometric evaluation of $200ng/cm^2$ PDGF-BB loaded bioresorbable membrane made from polyglycolic and polylactic acid. It was tested for its biocompatibility, ability to prevent epithelial downgrowth and amount of periodontal regeneration. Without membrane and PDGF-BB unloaded bioresorbable membrane were used as control. Healthy six beagle dogs were used. Each dog was anesthetized and buccal flaps were reflected in the mandibular and maxillary premolar areas. Buccal alveolar bone between the mesiobuccal and distobuccal line angles was surgically removed on the lower 2nd and 4th premolar in mandible, 2nd premolar in maxilla, to a level 4mm apical to the cementoenamel junction with creating a Class II buccal furcation defect for available space. Care was taken not to remove the root cementum layer and rubber impression materials were placed over each surgically created defect. Flaps were repositioned and sutured. Reconstructive surgery was performed 1 month after defect preparation. PDGF-BB loaded membranes and controls were randomly placed on maxillary 2nd premolars and mandibular 2nd and 4th premolars. Plaque control regimen was instituted with daily brushing with a 0.1% chlorhexidine digluconate during experimental periods. The animals were sacrificed 2 and 5 weeks after surgery and undecalcified specimens were prepared for histologic evaluation. The degree of coronal regrowth of new bone, new cementum and the amonut of new bone areas formed on the defected area of the PDGF-BB loaded membrnae turned superior to without membrane and drug unloaded membrane. Experimental membrane could prevent the epithelial downgrowth irrespective of drug loaded or not and showed good biocompatiblity, These results implicated that PDGF-BB loaded bioresorbable membrane could be highly useful tool for guided tissue regeneration of periodontal defects.

• 대한소아치과학회지
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• 제27권2호
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• pp.185-191
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• 2000

Streptococcus mutans가 치태를 형성할 때, 유산구균 1370 (Streptococcus lactis 1370)이 생산하는 수용성 글루캔이 영향을 미친다. 본 연구에서는 여러 인자에 의한 수용성 글루캔 형성 정도를 유산구균 1370배양 상청액의 흡광도로 측정하여 다음과 같은 결과를 얻었다. 1. 5% 자당이 첨가된 M17 broth에서 유산구균 1370 배양 상청액의 흡광도는 높고 Streptococcus mutans 배양 상청액의 흡광도는 낮았으나, 통계학적인 유의성은 없었다. 2. M17 broth에 10% 자당을 첨가하여 유산구균 1370을 배양할시 배양 상청액의 흡광도는 첨가하지 않을 때보다 높았다. (p<0.05), 배지 pH가 5에서보다 7에서 배양 상청액의 흡광도가 더 높았다(p<0.05). 3. $32^{\circ}C,\;37^{\circ}C,\;42^{\circ}C$중 $37^{\circ}C$에서 배양시 배양 상청액의 흡광도가 가장 높았으나 통계학적인 유의성이 없었고, 호기성 배양시보다 혐기성 배양시 배양 상청액의 흡광도가 더 높았다(p<0.05). 4. 배지의 $CaCl_2$ 농도가 1.0mM에서 (p<0.05), KCl 농도가 2.5mM에서 (p<0.05) 배양 상청 액의 흡광도가 높았다. 5. M17 broth에 5% 자당을 첨가한 배지에 유산구균 1370을 접종하여 배양한 배양 상청액과 배지를 1:3으로 가한 비커와이어 검사에서 Streptococcus mutans에 의하여 형성된 인공치태 무게는 5.6mg으로 5% 자당을 첨가하지 않을 때의 103.0mg에 비교하여 현저히 감소하였다 (p<0.05). 이상의 결과를 요약하면 유산구균 1370에 의한 수용성 글루캔의 형성은 자당이 함유된 배지나 세균 증식이 잘 되는 조건에서 증가하였으며, 수용성 글루캔이 Streptococcus mutans에 의한 인공치태 형성을 억제하였다.

• 대한소아치과학회지
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• 제35권1호
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• pp.83-91
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• 2008

치아 우식증은 구강 내 세균, 식이(음식물), 치아 및 타액 등의 숙주 요인이 복합적으로 작용하여 발생한다. 이 중 치아의 탈회에 대한 저항성을 높이거나, 구강 내 세균의 산생성능을 낮춤으로써 치아우식을 예방할 수 있는 대표적인 약제로 불소와 클로르헥시딘이 있다. 이 약제의 구강 내 적용방법으로 치아에 대한 부착력이 뛰어나고 환자의 협조의존도가 비교적 적은 바니쉬 형태가 최근 들어 널리 이용되고 있다. 본 연구는 불소와 클로르헥시딘 바니쉬의 항우식 효과를 생체 내에서 비교 평가하기 위하여, 구강 내 가철성 장치에 우치 시편을 식립하고 불소 바니쉬와 클로르헥시딘 바니쉬를 각각 도포하였다. 치태축적을 유도하여 법랑질 탈회를 통한 우식 유발 환경을 만들고 바니쉬 제제가 구강 내에서 우치 법랑질의 우식 예방에 미치는 효과를 평가하였다. 전자 현미분석 장치와 편광현미경을 사용하여 법랑질 표면의 Ca, P에 대한 정량적 변화를 분석하고 조직학적 관찰을 시행하여 다음과 같은 결론을 얻었다. 1. 인공 우식병소에 대한 편광현미경 관찰 결과, 불소 및 클로르헥시딘 바니쉬 를 도포한 군에서 대조군에 비해 법랑질 병소가 경미하게 나타남을 확인할 수 있었다. 2. 우식을 유발한 경우 Ca와 P의 감소량은 대조군에서 가장 높게 나타났고(P<0.05), 불소 바니쉬군에서는 Ca와 P의 유의한 감소가 나타나지 않았으며 클로르헥시딘 바니쉬군에서는 P만 유의한 감소를 보였다(P<0.05). 이상의 결과로 보아 불소 및 클로르헥시딘 바니쉬 제제의 사용이 항우식 효과를 나타낸 것으로 판단되나, 불소 바니쉬가 좀 더 우수한 효과를 나타낸 것으로 보인다.

• Restorative Dentistry and Endodontics
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• 제25권1호
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• pp.1-16
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• 2000

It is difficult to treat the endodontic apical perforation successfully. In this study, we hypothesized that the application of PDGF-BB and IGF-I into periapical perforation site may accelerate periapical healing and lead to bone deposition. And the specificity of osteonectin in periapical healing was investigated. The experiments were performed on the upper and lower 51 premolar teeth of 4 beagle dogs. The pulp chamber of each tooth was opened and the dental plaque was inserted into the canal for developing the periapical lesion for 5 weeks. Then, the roots were artificially perforated at the apex with the number 4 profile of .06 taper. In each step, standard periapical radiographs were taken to compare the size of lesion each other. The radiographs were scanned and analyzed by image analysis system. The mean and standard deviation of periradicular radiolucency ratios were calculated in each group. ANOVA was used for comparison. 51 premolars were grouped into 3 groups; control group, calcium hydroxide-treated group and calcium hydroxide plus growth factors-treated group. In the control group, the apical perforations were not sealed and obturated with gutta-percha and ZOE sealer by lateral condensation technique. In the experimental groups, the apical perforation were sealed with calcium hydroxide and with/without $4{\mu}g$ of PDGF-BB & IGF-I in cellulose gel and obturated by lateral condensation technique. Fluorescent bone markers were used to measure new bone formation. Following 2, 4, 12 weeks after experiment the dogs were sacrificed and histologic sections were prepared. Each tooth block including periapical lesion was sectioned mesiodistally. One half of the sections were decalcified with 6% nitric acid and processed by standard paraffin embedding technique. The sections were stained by hematoxylin and eosin, and immunostained for osteonectin. Histomorphometrical measurement of neoformed bone was performed using a light microscope. And the other half of the sections were prepared by undecalcified preparation, and confocal laser scanning microscopic investigations were done.