A.A. Del Bel Cury

In situ relationship between sucrose exposure and the composition of dental plaque

The aim of this study was to analyze the composition of dental plaque according to sucrose exposure. Twelve adult volunteers took part in this crossover study done in four phases of 28 days each. For each phase of the study, an acrylic resin appliance containing four human dental enamel blocks was constructed for each volunteer. A 20% sucrose solution was dripped onto the enamel blocks from 0 to 8 times/day. The volunteers were randomly assigned to the treatments. During the experimental period all the subjects used fluoride-free dentifrice, refrained from brushing the enamel blocks and drank water fluoridated at 0.70 ppm F. After each phase the concentrations of fluoride (F), calcium (Ca), phosphorus (P) and total carbohydrate were determined in dental plaque. Statistical analyses showed that frequent sucrose exposure significantly (p < 0.05) reduced the F, Ca and P concentrations in dental plaque, but increased the alkali-soluble carbohydrate concentration. The results suggest that the cariogenicity of dental plaque formed in the presence of sucrose cannot be attributed only to its higher porosity, but the lower inorganic concentration may also be important.

In situ Effect of Frequent Sucrose Exposure on Enamel Demineralization and on Plaque Composition after APF Application and F Dentifrice Use

Since the effect of the combination of methods of fluoride use on enamel demineralization and on plaque composition is not clearly established, this study examined the effect of the combination of acidulated phosphate fluoride (APF) application and F dentifrice on enamel demineralization and on plaque composition. In this crossover study, 16 volunteers, wearing a palatal appliance containing bovine enamel blocks, were subjected to 4 treatment groups: non-fluoridated dentifrice (PD), FD, APF+PD, and APF+FD. The APF was applied to the enamel before the 14-day experimental period. During the experimental period, test dentifrices were applied 3×/day, and a 20% sucrose solution was applied 4× and 8×/day by being dripped on the blocks. Although APF application was able either to increase F concentration in plaque or to reduce the % of mutans streptococci, its combination with F dentifrice use neither reduced enamel mineral loss nor changed any other measured plaque variable with respect to the FD group alone.

Effect of Frequency of Sucrose Exposure on Dental Biofilm Composition and Enamel Demineralization in the Presence of Fluoride

It has been suggested that enamel would resist higher frequencies of sucrose exposure if fluoride from water or dentifrice is being used. However, the effect of increasing frequencies of sugar on dental biofilm composition is not well known. Ten volunteers living in a fluoridated area wore palatal appliances bearing human enamel slabs during 14 days. The slabs were exposed to 20% sucrose solution 0 (control), 2, 4, 6, 8 or 10 times/day and the volunteers used fluoride dentifrice 3 times/day. Enamel demineralization was significantly greater than control for sucrose frequencies higher than 6 times/day. However, biofilm mass, total microbiota, total streptococci, lactobacilli counts and insoluble extracellular polysaccharide concentration increased, while Ca, Pi and F concentration in whole biofilm decreased significantly, with frequencies of sucrose exposure lower than 6 times/day. The findings confirm that fluoride can reduce enamel demineralization if sucrose consumption is not higher than 6 times/day, but changes in the biochemical and microbiological composition of the biofilm are observed with lower frequencies of sucrose use.

Effect of sucrose concentration on dental biofilm formed in situ and on enamel demineralization

The relationship between sucrose concentration and cariogenic potential was studied in situ. Adult volunteers wore intraoral palatal appliances containing human dental enamel blocks, which were extraorally submitted 8 times a day for 14 days, to the treatments: deionized distilled water and sucrose solutions from 1 to 40%. The biofilm formed was analyzed with respect to acidogenicity and biochemical composition; enamel demineralization was evaluated by microhardness. The results showed that 1% sucrose is less cariogenic than 5% or higher concentrations, although sucrose solution at 40% was still able to increase the concentration of insoluble polysaccharide in the biofilm formed. The findings suggest that the threshold of sucrose solution concentration for the formation of a cariogenic biofilm is 5%, which provided the same cariogenic potential as that observed for 10 and 20% sucrose solution.

Effect of Starch and Sucrose on Dental Biofilm Formation and on Root Dentine Demineralization

The cariogenicity of starch alone or in combination with sucrose is controversial and the effect on dentine demineralization and on the dental biofilm formed has not been explored under controlled conditions. A crossover, single-blind study was conducted in four steps of 14 days each, during which 11 volunteers wore palatal appliance containing 10 slabs of root dentine to which the following treatments were applied extraorally: 2% starch gel-like solution (starch group); 10% sucrose solution (sucrose group); a solution containing 2% starch and 10% sucrose (starch + sucrose group), or 2% starch solution followed by 10% sucrose solution (starch → sucrose group). On the 14th day of each phase the biofilms were collected for biochemical and microbiological analyses, and dentine demineralization was assessed by hardness. A higher demineralization was found in dentine exposed to sucrose and starch sucrose combinations than to starch alone (p < 0.01), but the sucrose-containing groups did not differ significantly from each other (p > 0.05). The concentrations of soluble and insoluble extracellular polysaccharides (EPS), and the proportion of insoluble EPS, were lower in the biofilm formed in presence of starch (p < 0.01) than in those formed in the presence of sucrose or sucrose/starch combinations; however, no significant difference was observed among the groups containing sucrose (p > 0.05). RNA was successfully isolated and purified from in situ biofilms and only biofilms formed in response to sucrose and starch/sucrose combinations showed detectable levels of gtfB and gtfC mRNA. The findings suggest that the combination of starch with sucrose may not be more cariogenic to dentine than sucrose alone.

Effect of Microleakage and Fluoride on Enamel-Dentine Demineralization around Restorations

There is no consensus about an association between microleakage and secondary caries, especially considering the presence of fluoride (F) at the tooth/restoration interface. Thus, a randomized, double-blind, crossover study was carried out to evaluate in situ the effect of microleakage on caries around enamel-dentine restorations in the presence of F from dental materials or dentifrice, either alone or in combination. In 4 phases of 14 days each, 14 volunteers wore palatal devices containing dental slabs restored with composite resin (CR) or resin-modified glass ionomer cement (GI). Restorations were made without leakage (L–), following the recommended adhesive procedures, or with leakage (L+), in the absence of adhesive procedures. Plaque-like biofilm (PLB) was left to accumulate on the restored slabs, which were exposed extraorally to a 20% sucrose solution 10×/day. The volunteers used a non-F (NF) or an F (FD) dentifrice 3×/day, depending on the experimental phase. No differences were found between L+ or L– restorations (p > 0.05). Higher demineralization in both enamel and dentine around CR restorations was observed under NF (p < 0.05). F concentration was higher in the fluid of PLB exposed to FD or formed onto GI restoration (p < 0.05). These results suggest that while microleakage does not affect caries development, GI or FD may maintain increased F levels in the PLB, thereby decreasing caries progression.

Effect of sucrose containing iron (II) on dental biofilm and enamel demineralization in situ

Since the effect of iron (Fe) on the cariogenicity of sucrose in humans is unexplored, this study assessed in situ the effect of Fe co-crystallized with sucrose (Fe-sucrose) topically applied in vitro on the acidogenicity, biochemical and microbiological composition of the dental biofilm formed in vivo and on the demineralization of the enamel. During two phases of 14 days each, 16 volunteers wore palatal appliances containing blocks of human enamel, which were submitted to four groups of separate treatments: (1) water; (2) 20% sucrose; (3) 20% (w/v) sucrose plus 18 µg Fe/ml, and (4) 20% (w/v) sucrose plus 70 µg Fe/ml. The solutions were dripped onto the blocks 8 times per day. The biofilms formed on the blocks were analyzed with respect to acidogenicity, biochemical and microbiological composition. Mineral loss was determined on enamel by surface and cross-sectional microhardness. Lower demineralization was found in the blocks subjected to Fe-sucrose (70 µg Fe/ml) than in those treated with sucrose (p < 0.05). This concentration of Fe also reduced significantly the populations of mutans streptococci in the biofilm formed on the blocks. In conclusion, our data suggest that Fe may reduce in situ the cariogenic potential of sucrose and the effect seems to be related to the reduction in the populations of mutans streptococci in the dental biofilm formed.

Ca, Pi, and F in the Fluid of Biofilm Formed under Sucrose

Calcium (Ca), inorganic phosphorus (Pi), and fluoride (F) concentrations are low in the whole plaque biofilm formed under exposure to sucrose. It was hypothesized that this would be reflected in the biofilm fluid, where these low values should greatly influence the de/remineralization process. Dental biofilms were formed in situ over enamel blocks mounted in palatal appliances and exposed 8 times/day to distilled water, glucose+fructose, or sucrose solutions for 14 days. While Ca, Pi, and F concentrations in the whole biofilms were significantly lower in the glucose+fructose and sucrose groups, no effect on biofilm fluid was observed, even after a cariogenic challenge. An increase in whole biofilm mineral ions was observed 24 hrs after the carbohydrate treatments were suspended, but this effect was also not observed in the fluid. These results suggest that there is a homeostatic mechanism that maintains biofilm fluid mineral ion concentration, regardless of its total concentration in the whole biofilm.

Effect of Sucrose on the Selection of Mutans Streptococci and Lactobacilli in Dental Biofilm Formedin situ

Microorganisms are selected in dental biofilm by the acidic environment created by sugar fermentation, but the effect of extracellular polysaccharide (EPS) on the counts of cariogenic bacteria is not clear. Dental biofilm was formed in situ for 13 days under exposure 8 times a day to distilled-deionized water, glucose + fructose or sucrose solutions. Mutans streptococci (MS) counts were not different among the groups, but lactobacilli (LB) were significantly higher in glucose + fructose and sucrose groups, without significant difference between them, irrespective of the higher insoluble EPS concentration in the sucrose biofilm matrix. The data suggest that exposure to sugar is more relevant for the predominance of LB in dental biofilm than for MS and that insoluble EPS does not change the counts of these microorganisms in the biofilm.

Effect of a calcium carbonate-based dentifrice on enamel demineralization in situ

Since the effect of calcium carbonate (CaCO3) based dentifrice on enamel demineralization is not clearly established, it was evaluated using the IEDT model described by Zero’s group in 1992. This study had a crossover design and 10 volunteers were submitted to 3 treatment groups: a negative control, brushing without dentifrice; an active control, brushing with silica-based dentifrice (SiO2/MFP group), and the experimental group, brushing with CaCO3-based dentifrice (CaCO3/MFP). Both dentifrices contained 1,500 µg F/g (w/w) as sodium monofluorophosphate (MFP). Enamel surface microhardness was determined in the dental blocks and the percentage change in relation to baseline was calculated. Fluoride uptake in enamel and its concentration in ‘test plaque’ were determined. The results showed that the dentifrice containing CaCO3/MFP was more effective than SiO2/MFP in reducing enamel demineralization (p < 0.05). A higher concentration of fluoride ion was found in ‘test plaque’ treated with CaCO3/MFP than in the negative control (p < 0.05). The results suggest that CaCO3 abrasive may enhance the effect of fluoride present in dentifrice on dental caries control.