An in vitro Microbial Model for Studying Secondary Caries Formation

Fontana, Margherita; Dunipace, Ann J.; Gregory, Richard L.; Noblitt, Timothy W.; Li, Y.; Park, K. K.; Stookey, George K.

doi:10.1159/000262146

Cited by 55 publications

(66 citation statements)

References 13 publications

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“…Thus, the inner lesion is the result of bacteria, fluids and hydrogen ions diffusing into the gap between restoration and cavity wall. Wall lesions were also observed in later examinations [4][5][6][7][8][9][10].…”

Section: Introductionmentioning

confidence: 95%

Isolated development of inner (wall) caries like lesions in a bacterial-based in vitro model

et al. 2009

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The study conducted in a bacterial-based in vitro caries model aimed to determine whether typical inner secondary caries lesions can be detected at cavity walls of restorations with selected gap widths when the development of outer lesions is inhibited. Sixty bovine tooth specimens were randomly assigned to the following groups: test group 50 (TG50; gap, 50 μm), test group 100 (TG100; gap, 100 μm), test group 250 (TG250; gap, 250 μm) and a control group (CG; gap, 250 μm). The outer tooth surface of the test group specimens was covered with an acidresistant varnish to inhibit the development of an outer caries lesion. After incubation in the caries model, the area of demineralization at the cavity wall was determined by confocal laser scanning microscopy. All test group specimens demonstrated only wall lesions. The CG specimens developed outer and wall lesions. The TG250 specimens showed significantly less wall lesion area compared to the CG (p<0.05). In the test groups, a statistically significant increase (p<0.05) in lesion area could be detected in enamel between TG50 and TG250 and in dentine between TG50 and TG100. In conclusion, the inner wall lesions of secondary caries can develop without the presence of outer lesions and therefore can be regarded as an entity on their own. The extent of independently developed wall lesions increased with gap width in the present setting.

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Section: Introductionmentioning

confidence: 95%

Isolated development of inner (wall) caries like lesions in a bacterial-based in vitro model

et al. 2009

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“…For each study, four groups of 12 human teeth specimens per group were treated for a 4-day test period in an in vitro microbial artificial-mouth caries model (1). The groups differed from each other in the presence (A1, A2, and A3) or absence (B1, negative control) of S. mutans (serotype c) and in the concentration of antibody used (Tables 1 and 2).…”

Section: In Vitro Microbial Caries Model Experiments (I) General Expmentioning

confidence: 99%

An In Vitro Microbial-Caries Model Used to Study the Efficacy of Antibodies toStreptococcus mutansSurface Proteins in Preventing Dental Caries

Fontana¹,

Buller²,

Dunipace³

et al. 2000

Clin Diagn Lab Immunol

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The first step for a pathogenic bacterium to initiate infection is via attachment (i.e., through surface determinants) to a suitable receptor. An in vitro microbial artificial-mouth model was used to test the efficacy of polyclonal antibodies to Streptococcus mutans cell surface proteins (CsAb) and a cell surface 59-kDa protein (59Ab) in preventing S. mutans colonization and carious lesion formation. In study 1, groups of 12 human teeth specimens were inoculated with S. mutans, which were incubated with different concentrations of CsAb (A1 [positive control], sterile saline, no antibody; A2, 0.007 mg of antibody protein/ml; and A3, 0.7 mg of antibody protein/ml) for 1 h at 37°C. The negative control group (B1) was not infected and was incubated with Trypticase soy broth (TSB) without dextrose supplemented with 5% sucrose (TSBS). In study 2, the same study design was used except that 59Ab was used instead of CsAb, normal rabbit serum was used in the positive control group (A1), and TSB supplemented with 1% glucose was used as the nutrient to control for sucrose-dependent colonization. All groups were exposed for 4 days to circulating cycles of TSBS and TSB (study 1 and study 2, respectively; 30 min each, three times per day) and a mineral washing solution (21 h per day). Prior to each nutrient cycle, 1 ml of the appropriate CsAb or 59Ab solution was administered to each group and allowed to mix for 30 min before cycling was resumed. Data obtained by confocal laser scanning microscopy demonstrated the presence of a significantly smaller (P < 0.05) lesion area and a smaller total lesion fluorescence in group A3 than in group A1 for both studies. In study 1, group A2 had significantly smaller values than A1 for lesion depth and area. There were no significant differences between groups A2 and A3 for lesion area or between groups A1 and A2 for total lesion fluorescence. In study 2, there were no significant differences among groups A1 and A2 for lesion depth or between groups A2 and A3 for all of the parameters studied. In both studies, there were no significant differences between S. mutans plaque CFU numbers among any of the groups. These studies demonstrated the efficacy of CsAb and 59Ab in reducing primary caries development in this model, although the underlying mechanism remains unclear.

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“…However, for fluoride to exert its cariostatic properties, it needs to diffuse through the biofilm to reach the enamel surface with the thickness of the biofilm being a rate-limiting parameter [5]. While biofilms have been successfully incorporated into laboratory models by various investigators [6][7][8] , studying the effect of thickness remains a challenge. Apart from certain devices used in situ that enable the growth of a biofilm to a confined thickness [9], under laboratory conditions, the biofilm is typically allowed to grow without controlling its thickness, although constant depth film fermenters can partially overcome this aspect [10].…”

Section: Introductionmentioning

confidence: 99%

Artificial biofilm thickness and salivary flow effects on fluoride efficacy – A model development study

Lippert¹,

Hara²,

Churchley³

2017

Res Rev Insights

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This laboratory model development study investigated the interaction between artificial biofilm thickness and salivary flow rate on fluoride-mediated prevention of enamel caries lesion formation. This 5-day pH cycling study on sound bovine enamel specimens utilized a continuous flow model and followed a 4 (agarose biofilm thickness-'no biofilm'/1/2/3mm)×2 (remineralizing solution flow rate-0.05/0.5ml/min)×2 (fluoride-0/383ppm as sodium fluoride) factorial design. Vickers surface microhardness change was the outcome measure. Data were analyzed with three-way ANOVA. The three-way interaction gel thickness×flow rate×fluoride concentration was significant (p=0.0006). 383ppm fluoride caused less softening than 0ppm regardless of gel thickness or flow rate. 0.5ml/min flow rate caused less softening than 0.05ml/min for 'no biofilm' and 1mm biofilm thickness regardless of fluoride concentration, for 2 and 3mm with 0ppm F but not for 383ppm F. For 0.05ml/min, softening was reduced as gel thickness increased from 'no biofilm'-1-2mm, but not from 2-3mm. For 0.5ml/min, 'no biofilm' caused more softening than 1, 2, and 3mm, but 1, 2, and 3mm were not different from each other for both 0 and 383ppm F. The present findings suggest that the efficacy of fluoride in preventing enamel demineralization is affected by both biofilm thickness and salivary flow rate, with both thicker biofilms and higher flow rate resulting in less demineralization.

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An in vitro Microbial Model for Studying Secondary Caries Formation

Cited by 55 publications

References 13 publications

Isolated development of inner (wall) caries like lesions in a bacterial-based in vitro model

Isolated development of inner (wall) caries like lesions in a bacterial-based in vitro model

An In Vitro Microbial-Caries Model Used to Study the Efficacy of Antibodies toStreptococcus mutansSurface Proteins in Preventing Dental Caries

Artificial biofilm thickness and salivary flow effects on fluoride efficacy – A model development study

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