Antônio Pedro Ricomini Filho

Efficacy of denture cleansers on Candida spp. biofilm formed on polyamide and polymethyl methacrylate resins

STATEMENT OF PROBLEM Although new materials have emerged as options to fabricate removable dental prostheses, the development of Candida biofilms on these materials and the effectiveness of methods to control these pathogenic biofilms are poorly understood. PURPOSE The purpose of this study was to evaluate the efficacy of denture cleansers on Candida single- and dual-species biofilms formed on polyamide resin. MATERIAL AND METHODS Polymethyl methacrylate (PMMA) resin (Acron MC) and polyamide resin (Flexite M.P.) specimens (n=116) were prepared, and their surface roughness was standardized (0.34 ±0.02 μm). Surface free energy (SFE) was measured for some specimens (n=20 per resin), while the remainder were randomly divided by lottery into 24 groups (n=8) for biofilm assay. C. albicans and/or C. glabrata biofilm was formed for 72 hours, and then specimens were treated with an enzymatic cleanser solution (Polident 3 Minutes), a cleanser solution (Corega Tabs), or 0.5% sodium hypochlorite (NaOCl) solution. Water served as the negative control. Remaining adherent microorganisms were removed from the treated specimens by ultrasonic waves, and colony-forming units (CFU) of each microorganism were calculated. SFE data were analyzed by 1-way ANOVA, and Candida species data were analyzed by 3-way ANOVA followed by the Tukey-Kramer test (P=.05). RESULTS All tested biofilms displayed significantly higher growth on polyamide resin (P<.001), which presented the lowest SFE. Denture cleansers significantly decreased Candida levels; however, the 0.5% NaOCl solution was the only effective cleanser. C. glabrata revealed significantly higher CFU counts under all experimental conditions (P<.001). CONCLUSIONS The highest Candida spp. biofilm growth was shown to occur on polyamide resin when compared with PMMA. Denture cleansers were able to remove Candida spp. biofilm formed on both denture base resins.

Preload loss and bacterial penetration on different implant-abutment connection systems

Preload loss can favor the occurrence of implant-abutment interface misfit, and bacterial colonization at this interface may lead to implant failure. The aim of this study was to evaluate the preload loss and bacterial penetration through the implant-abutment interface of conical and external hexagon connection systems subjected to thermal cycling and mechanical fatigue (TM). Four different implant-abutment connection systems were evaluated (n=6): external hexagon with universal post, Morse taper with universal post, Morse taper with universal post through bolt, and locking taper with standard abutment. The assemblies (implant-abutment) were subjected to a thermal cycling regimen (1,000 cycles of 5 degrees C and 55 degrees C) and to mechanical fatigue (1.0 million cycles, 1.0 Hz, 120 N). The assemblies were immersed in Tryptic Soy + Yeast Extract broth containing Streptococcus sanguinis and incubated at 37 degrees C and 10% CO(2) for 72 h. Detorque values were recorded. The bacterial penetration was assessed and the abutments were observed by scanning electron microscopy. The preload data were analyzed statistically by two-way ANOVA and Tukeys test at 5% significance level. All screw abutment systems showed significantly higher (p<0.05) detorque values when subjected to TM and all conical systems presented bacterial penetration. The results show no relationship between the preload loss and the bacterial penetration.

Fluoride concentration in the top-selling Brazilian toothpastes purchased at different regions

To be relevant in terms of public health, widely-used toothpastes should have at least 1,000 ppm of soluble fluoride (F) concentration. Thus, the concentrations of total fluoride (TF) and total soluble fluoride (TSF) in the top-selling Brazilian toothpastes were evaluated. Samples (n=3) from toothpastes Colgate Anti-cáries(®), Colgate Total 12 Clean Mint(®), Colgate Tripla Ação Menta Original(®), Colgate Tripla Ação Menta Suave(®) and Sorriso Dentes Brancos(®) were obtained from each of the five regions of the country. The concentrations of TF and TSF were analyzed with ion-specific electrode calibrated with F standards and the results were expressed in ppm (µg F/g). All toothpastes showed TF concentration lower than 1,500 ppm F (1,388.2 ± 25.8 to 1,483.2 ± 98.2). The TSF values were higher than 1,000 ppm F and ranged from 1,035.5 ± 61.5 to 1,221.8 ± 35.2 for calcium carbonate/monofluorophosphate-based toothpastes and from 1,455.6 ± 12.5 to 1,543.0 ± 147.3 for silica/sodium fluoride-based toothpaste. Top-selling Brazilian toothpastes presented available fluoride concentration to control caries regardless of the region where they are purchased.

Polytetrafluorethylene added to acrylic resins: mechanical properties

The addition of different polymers, such as polytetrafluorethylene (PTFE), to denture base resins could be an option to modify acrylic resin mechanical properties. This study evaluated the surface hardness, impact and flexural strength, flexural modulus and peak load of 2 acrylic resins, one subjected to a long and another subjected to a short polymerization cycle, which were prepared with or without the addition of 2% PTFE. Four groups were formed according to the polymerization cycle and addition or not of PTFE. Forty specimens were prepared for each test (10 per group) with the following dimensions: hardness (30 mm diameter x 5 mm thick), impact strength (50 x 6 x 4 mm) and flexural strength (64 x 10 x 3.3 mm) test. The results of the flexural strength test allowed calculating flexural modulus and peak of load values. All tests were performed in accordance with the ISO 1567:1999 standard. Data were analyzed statistically by ANOVA and Tukeys test with the level of significance set at 5%. No statistically significant differences (p>0.05) were found for surface hardness. Flexural strength, impact strength and peak load were significantly higher (p<0.05) for resins without added PTFE. The flexural modulus of the acrylic resin with incorporated 2% PTFE polymerized by long cycle was significantly higher (p<0.05) than that of the other resins. Within the limits of this study, it may be concluded that the addition of PTFE did not improve the mechanical properties of the evaluated acrylic resins.

Effect of daily use of an enzymatic denture cleanser on Candida albicans biofilms formed on polyamide and poly(methyl methacrylate) resins: an in vitro study.

STATEMENT OF PROBLEM Candida biofilms on denture surfaces are substantially reduced after a single immersion in denture cleanser. However, whether this effect is maintained when dentures are immersed in cleanser daily is unclear. PURPOSE The purpose of this study was to evaluate the effect of the daily use of enzymatic cleanser on Candida albicans biofilms on denture base materials. MATERIAL AND METHODS The surfaces of polyamide and poly(methyl methacrylate) resin specimens (n=54) were standardized and divided into 12 groups (n=9 per group), according to study factors (material type, treatment type, and periods of treatment). Candida albicans biofilms were allowed to form over 72 hours, after which the specimens were treated with enzymatic cleanser once daily for 1, 4, or 7 days. Thereafter, residual biofilm was ultrasonically removed and analyzed for viable cells (colony forming units/mm(2)) and enzymatic activity (phospholipase, aspartyl-protease, and hemolysin). Factors that interfered with the response variables were analyzed by 3-way ANOVA with the Holm-Sidak multiple comparison method (α=.05). RESULTS Polyamide resin presented more viable cells of Candida albicans (P<.001) for both the evaluated treatment types and periods. Although enzymatic cleansing significantly (P<.001) reduced viable cells, daily use did not maintain this reduction (P<.001). Phospholipase activity significantly increased with time (P<.001) for both materials and treatments. However, poly(methyl methacrylate) based resin (P<.001) and enzymatic cleansing treatment (P<.001) contributed to lower phospholipase activity. Aspartyl-protease and hemolysin activities were not influenced by study factors (P>.05). CONCLUSIONS Although daily use of an enzymatic cleanser reduced the number of viable cells and phospholipase activity, this treatment was not effective against residual biofilm over time.

Biofilm extracellular polysaccharides degradation during starvation and enamel demineralization

This study was conducted to evaluate if extracellular polysaccharides (EPS) are used by Streptococcus mutans (Sm) biofilm during night starvation, contributing to enamel demineralization increasing occurred during daily sugar exposure. Sm biofilms were formed during 5 days on bovine enamel slabs of known surface hardness (SH). The biofilms were exposed to sucrose 10% or glucose + fructose 10.5% (carbohydrates that differ on EPS formation), 8x/day but were maintained in starvation during the night. Biofilm samples were harvested during two moments, on the end of the 4th day and in the morning of the 5th day, conditions of sugar abundance and starvation, respectively. The slabs were also collected to evaluate the percentage of surface hardness loss (%SHL). The biofilms were analyzed for EPS soluble and insoluble and intracellular polysaccharides (IPS), viable bacteria (CFU), biofilm architecture and biomass. pH, calcium and acid concentration were determined in the culture medium. The data were analyzed by two-way ANOVA followed by Tukey’s test or Students t-test. The effect of the factor carbohydrate treatment for polysaccharide analysis was significant (p < 0.05) but not the harvest moment (p > 0.05). Larger amounts of soluble and insoluble EPS and IPS were formed in the sucrose group when compared to glucose + fructose group (p < 0.05), but they were not metabolized during starvation time (S-EPS, p = 0.93; I-EPS, p = 0.11; and IPS = 0.96). Greater enamel %SHL was also found for the sucrose group (p < 0.05) but the demineralization did not increase during starvation (p = 0.09). In conclusion, the findings suggest that EPS metabolization by S. mutans during night starvation do not contribute to increase enamel demineralization occurred during the daily abundance of sugar.

Fluoride Penetration and Clearance Are Higher in Exopolysaccharide-Containing Bacterial Pellets

Extracellular polysaccharides (EPS) could increase the penetration of fluoride through dental biofilm, reducing its cariogenicity. We measured the concentration of fluoride in EPS-containing (EPS+) or not-containing (EPS–) Streptococcus mutans bacterial pellets resembling test biofilms, before and up to 60 min after a 0.05% NaF rinse in situ. Fluoride penetration and clearance were higher in EPS+ bacterial pellets. The data suggest that EPS enhances fluoride penetration, but also accelerates fluoride clearance from dental biofilms.

Starch Combined with Sucrose Provokes Greater Root Dentine Demineralization than Sucrose Alone

Since there is no consensus about whether starch increases the cariogenic potential of sucrose, we used a validated 3-species biofilm model to evaluate if starch combined with sucrose provokes higher root dentine demineralization than sucrose alone. Biofilms (n = 18) composed by Streptococcus mutans (the most cariogenic bacteria), Actinomces naeslundii (which has amylolytic activity), and Streptococcus gordonii (which binds salivary amylase) were formed on root dentine slabs under exposure 8 ×/day to one of the following treatments: 0.9% NaCl, 1% starch, 10% sucrose, or a combination of 1% starch and 10% sucrose. Before each treatment, biofilms were pretreated with human whole saliva for 1 min. The pH of the culture medium was measured daily as an indicator of biofilm acidogenicity. After 96 h of growth, the biofilms were collected, and the biomass, bacteria viability, and polysaccharides were analyzed. Dentine demineralization was assessed by surface hardness loss (% SHL). Biofilm bioarchitecture was analyzed using confocal laser scanning microscopy. Treatment with a starch and sucrose combination provoked higher (p = 0.01) dentine demineralization than sucrose alone (% SHL = 53.2 ± 7.0 vs. 43.2 ± 8.7). This was supported by lower pH values (p = 0.007) of the culture medium after daily exposure to the starch and sucrose combination compared with sucrose (4.89 ± 0.29 vs. 5.19 ± 0.32). Microbiological and biochemical findings did not differ between biofilms treated with the combination of starch and sucrose and sucrose alone (p > 0.05). Our findings give support to the hypothesis that a starch and sucrose combination is more cariogenic for root dentine than sucrose alone.