Dynamics and metabolic profile of oral keratinocytes (NOK-si) and Candida albicans after interaction in co-culture.

Understanding the interaction between oral keratinocytes (NOK-si) and Candida albicans is fundamental for the development of prevention strategies and new therapies for oral candidiasis. This study evaluated the dynamics and metabolic profile of these cells growing in co-culture by means of cell metabolism, number of CFU ml-1, and production of enzymes, cytokines, and metabolites. The data were analyzed by ANOVAs and post hoc tests (α = 0.05). In co-cultures, there were significant decreases in the cell metabolism of NOK-si and C. albicans and increases in the CFU ml-1 values of C. albicans biofilm. There were also significant increases in the production of cytokines by NOK-si and proteinase by C. albicans biofilm after their interaction. The metabolic balance of the main metabolites, amino acids, and extracellular and intracellular metabolites was shifted in favor of the co-cultures, while aromatic alcohols were secreted in higher amounts by the biofilm of C. albicans. It was concluded that the interaction of cells in co-culture influenced their dynamics over time.

Long-Term Effect of Daily Chemical Disinfection on Surface Topography and Candida Albicans Biofilm Formation on Denture Base and Reline Acrylic Resins.

PURPOSE: This study investigated the effect of long-term daily chemical disinfection on the topographic and Candida albicans biofilm formation on a denture base resin and a reline acrylic resin. MATERIAL AND METHODS: Circular samples (14 × 1.2 mm) were fabricated from a denture base (Vipi Wave) and reline acrylic resins (Tokuyama Rebase Fast II). Samples were kept in 50 ml of distilled water (48 h at 37°C). Subsequently, the samples were immersed in five different solutions: 0.5% sodium hypochlorite; 3.8% sodium perborate; 2% chlorhexidine gluconate; apple vinegar containing 4% maleic acid; and distilled water (control group). The specimen was immersed in the solutions for 8 h daily and transferred to distilled water at 37°C for more 16 h. The surface topographic and Candida albicans (ATCC 90028) biofilm formation were evaluated at baseline (before chemical disinfection) and after 1, 3 and 6 months of immersion. The surface topographic was evaluated by arithmetical roughness average (Ra) and scanning electron microscope (SEM), while the biofilm formation was evaluated by colony-forming units (CFU/ml) method and Alamar Blue assay (cell metabolism). The results were evaluated by three-way analysis of variance (ANOVAs) and post-hoc tests (α = 0.05). RESULTS: The results showed statistically significant effects from the type of acrylic resin (p = 0.029) and time (p <0.001) on the roughness of the specimen. In general, the reline resin had higher roughness than the denture base resin. In addition, the roughness of the samples after 1, 3 and 6 months of immersion in the cleaning solutions was higher than at baseline. In relation to the microbiological assays, there were no statistically significant differences (p >0.055) in the CFU/ml values of the biofilms among the different resins, periods of time and cleaning solutions. Considering the metabolism of the cells within the biofilms, the results showed that, at baseline, it was statistically significantly higher (p <0.05) than after 1, 3 and 6 months of storage. The SEM images showed that all disinfectant solutions provided surface changes of both acrylic resins (base and reline) after 1, 3 and 6 months of immersion. CONCLUSIONS: The roughness of both acrylic resins was affected by the disinfection in all cleaning agents, increasing over time, and this effect was more evident in the reline acrylic resin group. This surface change was also observed in the SEM images. While the number of cells within the biofilms was not affected by immersion in the cleaning agents, their metabolism was lower after 1, 3 and 6 months of immersion.

Antimicrobial photodynamic therapy effectiveness against susceptible and methicillin-resistant Staphylococcus aureus biofilms.

BACKGROUND: Staphylococcus aureus have a great ability to become rapidly resistant to conventional antimicrobial therapies. This study evaluated the efficacy of antimicrobial photodynamic therapy (aPDT) mediated by Curcumin (Cur) and light-emitting diode (LED) in the inactivation of biofilms of methicillin susceptible and resistant S. aureus (MSSA and MRSA, respectively). METHODS: Biofilms were treated with Cur (20, 40 or 80 µM) and illuminated with LED source (455 ± 3 nm; 5.28 J/cm2) (aPDT groups), or treated either with Cur or LED only. Other samples were not exposed to Cur or LED (negative control). The biofilms viability after all experimental conditions were evaluated by counting the number of colonies (CFU/mL) and XTT assay. Additional samples were also evaluated by LIVE/DEAD® staining using confocal laser scanning microscopy (CLSM). Data were analyzed by ANOVAs followed by the Games-Howell post hoc test (α = 0.05). RESULTS: For both strains, all aPDT groups significantly reduced both CFU/mL and metabolic activity of biofilms compared to the negative control (p < 0.001). The results were enhanced when 80 µM of Cur was used. CLSM images showed that both bacteria biofilms submitted to aPDT had a large number of red-stained colonies, especially at aPDT80. In general, MRSA biofilms tended to be less susceptible to aPDT than MSSA biofilms. CONCLUSIONS: It can be concluded that aPDT mediated by Cur and LED was an efficient method to inactivate 48 -h biofilms of both S. aureus strains.

The influence of Cisplatin-based chemotherapy on the osseointegration of dental implants: An in vivo mechanical and histometrical study.

OBJECTIVES: To evaluate the effect of Cisplatin on bone repair and mineralization around implants and on the mechanical properties of bone tissue. MATERIALS AND METHODS: Forty-three Wistar rats were randomly divided into two groups: Cisplatin (CIS, medication) and control (CTL, placebo solution), administered once a week for 4 weeks. After 4 weeks, implants were installed in both tibiae metaphysis. After 30 and 60 days, the animals were sacrificed and their femurs and tibiae were removed. Femurs were subjected to mechanical tests and tibiae for removal torque, arrangement and distribution of collagen fibers, morphometrical analyses (bone tissue in contact with the implant surface [BIC] and areal fraction between implant threads occupied by bone tissue [BAFO]) and scanning electron microscopy to calcium and calcium/phosphorus analysis. Data were analyzed by ANOVA or MANOVA, and Tukey or Games-Howell post hoc tests, respectively (α = 0.05). RESULTS: The CTL specimens had significantly higher values (0.0001 ≤ p≤0.036) of strength (N), removal torque (N/cm2 ), %BIC, and %BAFO than CIS specimens, being their best results at day 60. No significant differences were found among the groups regarding the values of deformation, percentage of calcium, and calcium/phosphorus ratio. In CIS groups, there was a reduction in the organization of collagen at the bone/implant interface, resulting in a trabecular bone with thin trabeculae and birefringent collagen and irregular arrangement. CONCLUSIONS AND CLINICAL IMPLICATIONS: Cisplatin interfered negatively in the repair and mineralization around dental implants, as well as on the quality of the bone tissue, mainly in the period of 30 days after the implant placement.

Physical properties of artificial teeth after immersion in liquid disinfectant soaps.

PURPOSE: To evaluate the hardness, roughness and color stability of artificial teeth after immersion in liquid disinfectant soaps. METHODS: Artificial teeth (Vipi Dent Plus, ArtiPlus and Biolux) were divided into four groups (n=15), according to the type of immersion solution: distilled water/control group (DW); liquid disinfectant soap Dettol (SD); liquid disinfectant soap Protex (SP); and liquid disinfectant soap Lifebuoy (SL). The immersion cycles occurred every day, for 8 hours at room temperature in each disinfectant solution, following immersion in distilled water for 16 hours at 37°C. All solutions were changed daily. Properties were evaluated after 0, 7, 14, 21 and 28 days of immersion. The data were analyzed with a mixed three-way ANOVA followed by the Bonferroni post-hoc test (α= 0.05). RESULTS: Vipi teeth presented significant reduction (P< 0.05) in hardness and roughness prior to 7 days of immersion in all solutions, including control group. These values, in general, were maintained during the 28 days. Biolux teeth, in general, did not present significant changes in hardness prior to immersion in any of the time intervals. The roughness of these teeth increased after 21 and 28 days of immersion (P< 0.05) in all the solutions. ArtiPlus teeth maintained stable roughness and hardness during the assessment period, regardless of the type of soap used. Color alterations were considered clinically acceptable. The liquid soaps may be an alternative for the disinfection of partial or total removable dentures. CLINICAL SIGNIFICANCE: The liquid disinfectant soaps tested did not significantly alter the hardness, roughness and color stability of the artificial teeth tested and may be an alternative for the disinfection of partial or total removable dentures.

Effect of Chloroaluminium phthalocyanine in cationic nanoemulsion on photoinactivation of multispecies biofilm.

BACKGROUND: Photosensitizers in nanocarriers have been investigated for antimicrobial Photodynamic Therapy (aPDT). However, most studies are focused against microorganisms in planktonic or monospecies biofilm. Thus, this in vitro study evaluated the effect of aPDT using Chloroaluminium phthalocyanine (ClAlPc) in cationic nanoemulsion (NE) against Candida albicans, Candida glabrata and Streptococcus mutans grown as multispecies biofilm. METHODS: Standard suspensions of each microorganism were added into wells of a microtiter plate for biofilm growth for 48 h in a candle jar. The biofilms were incubated with ClAlPc in cationic NE at 31.8 µM for 30 min and illuminated with red light fluence of 39.3 J/cm2 (P+L+ group). Additional samples were treated only with photosensitizer (P+L-) or red light (P-L+) or neither (P-L-, control group). aPDT efficacy was assessed by colony quantification, biofilm's metabolic activity, total biomass, and confocal microscopy. Data were analyzed by ANOVA/Welch and post-hoc Tukey/Games-Howell tests (α = 0.05). RESULTS: aPDT (P+L+) reduced the colony count in 1.30 to 2.24 lg10 and the metabolic activity in 53.7% compared with the control group (P-L-). The total biomass showed no statistical difference among the groups. The confocal microscopy analyzes showed uptake of the PS in the biofilm, and dead cells were observed in the biofilm treated with aPDT. CONCLUSION: aPDT mediated by ClAlPc in cationic NE promoted photoinactivation of the multispecies biofilm, which was confirmed by colony quantification, metabolic activity, and confocal microscopy. However, the total biomass of the biofilm was not affected by the treatment.

Cytotoxic potential of denture base and reline acrylic resins after immersion in disinfectant solutions.

STATEMENT OF PROBLEM: The daily immersion of dentures in disinfectant solutions can cause the incorporation of toxic substances in the acrylic resins, and studies evaluating the cumulative effect of disinfectant solutions on cell culture are lacking. PURPOSE: The purpose of this in vitro study was to evaluate the cytotoxic potential of cell cultures of denture base and reline acrylic resins after immersion in disinfectant solutions. MATERIAL AND METHODS: Disk-shaped specimens (14×1.2 mm) were obtained and divided into groups (n=9) according to the disinfectant solutions (distilled water [control], 2% chlorhexidine digluconate, 3.8% sodium perborate, 0.5% sodium hypochlorite, and apple vinegar) and to the storage period (0, 1, 3, and 6 months). Solutions were changed daily. After the different storage periods, specimens were immersed in culture medium for 24 hours, and extracts were obtained. Human keratinocytes were cultivated, and the cellular metabolism was evaluated by using Alamar Blue. Data were submitted to 3-way analysis of variance and Games-Howell post hoc tests (α=.05). RESULTS: Both of the acrylic resins tested showed similar biocompatibility properties after immersion in different solutions (P=.400). Immersion in distilled water, 3.8% sodium perborate, and 0.5% sodium hypochlorite did not affect the cellular metabolism of the keratinocytes (P>.05), regardless of the immersion period and the type of acrylic resin (P>.05). Immersion in 2% chlorhexidine digluconate or apple vinegar resulted in high cytotoxicity over time, until the third month (P<.05), after which no changes were observed (P>.05). CONCLUSIONS: The type of acrylic resin (base or reline) had no significant effect on the viability of cells. Vinegar and chlorhexidine digluconate solutions increased in cytotoxic effect over time, and were strongly cytotoxic after 6 months of immersion. Sodium perborate and sodium hypochlorite were noncytotoxic in all periods of time tested.

Curcumin-mediated anti-microbial photodynamic therapy against Candida dubliniensis biofilms.

The purpose of this study was to evaluate the effectiveness of anti-microbial photodynamic therapy (aPDT) mediated by curcumin (Cur) associated with LED light against biofilms of Candida dubliniensis, and further, investigate cellular uptake and drug penetration through the biofilms under confocal laser scanning microscopy (CLSM). Four C. dubliniensis strains were tested: three clinical isolates from HIV-positive patients and one reference strain (CBS 7987). Biofilms were treated with three Cur concentrations (20.0, 30.0, and 40.0 µM). All samples were incubated in the dark for 20 min and exposed to a 5.28 J/cm2 of LED light fluence. Additional samples of each strain were treated either with Cur or LED light only. Control samples had neither Cur nor light. After aPDT, results were read using the XTT salt reduction method. The data were statistically analyzed by two-way ANOVA followed by Games-Howell post-hoc test (α = 0.05). Confocal laser scanning microscopy was used to verify both the uptake of Cur by yeast cells and its penetration through the biofilm. The results showed that aPDT promoted significant reduction on the metabolism of the biofilm-organized cells of C. dubliniensis. Further, while Cur was rapidly taken up by C. dubliniensis cells, a longer time interval was required to allow Cur penetration into biofilm cells. Based on these results, aPDT associating LED and Cur presents promising potential on fungal control of biofilms of C. dubliniensis.

Encapsulation of curcumin in polymeric nanoparticles for antimicrobial Photodynamic Therapy.

Curcumin (CUR) has been used as photosensitizer in antimicrobial Photodynamic Therapy (aPDT). However its poor water solubility, instability, and scarce bioavalibility hinder its in vivo application. The aim of this study was to synthesize curcumin in polymeric nanoparticles (NP) and to evaluate their antimicrobial photodynamic effect and cytoxicity. CUR in anionic and cationic NP was synthesized using polylactic acid and dextran sulfate by the nanoprecipitation method. For cationic NP, cetyltrimethylammonium bromide was added. CUR-NP were characterized by physicochemical properties, photodegradation, encapsulation efficiency and release of curcumin from nanoparticles. CUR-NP was compared with free CUR in 10% dimethyl sulfoxide (DMSO) as a photosensitizer for aPDT against planktonic and biofilms (mono-, dual- and triple-species) cultures of Streptococcus mutans, Candida albicans and Methicillin-Resistant Staphylococcus aureus. The cytotoxicity effect of formulations was evaluated on keratinocytes. Data were analysed by parametric (ANOVA) and non-parametric (Kruskal-Wallis) tests (α = 0.05). CUR-NP showed alteration in the physicochemical properties along time, photodegradation similar to free curcumin, encapsulation efficiency up to 67%, and 96% of release after 48h. After aPDT planktonic cultures showed reductions from 0.78 log10 to complete eradication, while biofilms showed no antimicrobial effect or reductions up to 4.44 log10. Anionic CUR-NP showed reduced photoinactivation of biofilms. Cationic CUR-NP showed microbicidal effect even in absence of light. Anionic formulations showed no cytotoxic effect compared with free CUR and cationic CUR-NP and NP. The synthesized formulations improved the water solubility of CUR, showed higher antimicrobial photodynamic effect for planktonic cultures than for biofilms, and the encapsulation of CUR in anionic NP reduced the cytotoxicity of 10% DMSO used for free CUR.

Reliability of the agar based method to assess the production of degradative enzymes in clinical isolates of Candida albicans.

The aim of this study was to establish a reproducible protocol using the methodology of hyaline zones around the colonies on specific agar plates for phospholipase and proteinase production. This was an in vitro double-blind experiment, in which the dependent variables were the enzymatic activity measurements (Pz) for the production of phospholipase (Pz-ph) and the production of secreted aspartyl proteinases (Pz-sap). Three independent variables give rise to different measurement protocols. All measurements were carried out at two different moments by four examiners (E1, E2, E3, and E4). The minimum sample size was 30 Candida albicans clinical isolates. Specific agar plates for phospholipase and SAPs production were prepared according the literature. The intra-and inter-examiner reproducibility for each protocol was estimated using the Intraclass Correlation Coefficient (ICC) and its confidence interval (95% CI). Based on the results obtained for both phospholipase and SAPs, there appears to be no consensus on the protocol chosen for each particular examiner. Measuring the colonies in triplicate may be the main factor associated with the increase in measurement accuracy and should therefore take precedence over measuring only one colony. When only one examiner is responsible for taking measurements, a standard protocol should be put in place and the statistical calibration of this researcher should be done prior to data collection. However, if two or more researchers are involved in the assessment of agar plates, our results suggest that the protocols using software to undertake plate reading is preferred.

Dynamics of biofilm formation and the interaction between Candida albicans and methicillin-susceptible (MSSA) and -resistant Staphylococcus aureus (MRSA).

Polymicrobial biofilms are an understudied and a clinically relevant problem. This study evaluates the interaction between C. albicans, and methicillin- susceptible (MSSA) and resistant (MRSA) S. aureus growing in single- and dual-species biofilms. Single and dual species adhesion (90 min) and biofilms (12, 24, and 48 h) were evaluated by complementary methods: counting colony-forming units (CFU mL-1), XTT-reduction, and crystal violet staining (CV). The secretion of hydrolytic enzymes by the 48 h biofilms was also evaluated using fluorimetric kits. Scanning electron microscopy (SEM) was used to assess biofilm structure. The results from quantification assays were compared using two-way ANOVAs with Tukey post-hoc tests, while data from enzymatic activities were analyzed by one-way Welch-ANOVA followed by Games-Howell post hoc test (α = 0.05). C. albicans, MSSA and MRSA were able to adhere and to form biofilm in both single or mixed cultures. In general, all microorganisms in both growth conditions showed a gradual increase in the number of cells and metabolic activity over time, reaching peak values between 12 h and 48 h (ρ<0.05). C. albicans single- and dual-biofilms had significantly higher total biomass values (ρ<0.05) than single biofilms of bacteria. Except for single MRSA biofilms, all microorganisms in both growth conditions secreted proteinase and phospholipase-C. SEM images revealed extensive adherence of bacteria to hyphal elements of C. albicans. C. albicans, MSSA, and MRSA can co-exist in biofilms without antagonism and in an apparent synergistic effect, with bacteria cells preferentially associated to C. albicans hyphal forms.

In vitro evaluation of the enzymatic activity profile of non-albicans Candida species isolated from patients with oral candidiasis with or without diabetes.

OBJECTIVE: To evaluate the expression of phospholipase (PL) and secreted aspartyl proteinase (SAP) by Candida glabrata and C tropicalis obtained from the denture biofilms of healthy participants (16 isolates), patients with oral candidiasis with diabetes (10 isolates), and patients with oral candidiasis without diabetes (25 isolates). STUDY DESIGN: After incubation, the supernatants and pellets of the isolates were used for the enzymatic assays and quantification of colony-forming units (CFU), respectively. Colorimetric tests were used with phosphatidylcholine as a substrate for PL and azocasein as a substrate for SAP, and the absorbances of the samples were measured. Enzymatic rates were calculated, and values were normalized by CFU. Results were analyzed with factorial analyses of variance (α = .05). RESULTS: C tropicalis and C glabrata were proteolytic and phospholipolytic. The clinical sources of isolates had no significant effect on the enzymatic activities (P > .05). C tropicalis had significantly higher enzymatic activity for both PL and SAP (P < .001) than did C glabrata. CONCLUSIONS: C tropicalis isolates produced significantly higher amounts of both enzymes than did the C glabrata isolates.

Enzymatic activity profile of a Brazilian culture collection of Candida albicans isolated from diabetics and non-diabetics with oral candidiasis.

The secretion of hydrolytic enzymes is a fundamental virulence factor of Candida albicans to develop disease. The objective of this study was to characterise the virulence of 148 clinical isolates of C. albicans from oral candidiasis by assessing the expression of phospholipase (PL) and secreted aspartyl proteinase (SAP). Isolates were obtained from healthy subjects (HS) and diabetics (DOC) and non-diabetics with oral candidiasis (NDOC). An aliquot (5 µl) of each cell suspension was inoculated on PL and SAP agar plates and incubated. Enzymes secretion was detected by the formation of an opaque halo around the colonies and enzymatic activity (PZ) was determined by the ratio between colony diameter and colony diameter plus the halo zone. Statistical comparisons were made by a one-way anova followed by Tukey's post hoc test (α = 0.05). The clinical sources of C. albicans had significant effect (P < 0.001) on the PZ values of both enzymes. For PL, clinical isolates from NDOC and DOC had highest enzymatic activity than those from HS (P < 0.05), with no significant differences between them (P = 0.506). For SAP, C. albicans from NDOC showed the lower enzymatic activity (P < 0.001). There were no significant differences between isolates from HS and DOC (P = 0.7051). C. albicans isolates from NDOC and DOC patients showed an increased production of PL.

Susceptibility profile of a Brazilian yeast stock collection of Candida species isolated from subjects with Candida-associated denture stomatitis with or without diabetes.

OBJECTIVE: This study investigated the susceptibility of 198 clinical isolates of Candida species against caspofungin, amphotericin B, itraconazole, and fluconazole. STUDY DESIGN: Suspensions of the microorganisms were spread on Roswell Park Memorial Institute (RPMI) agar plates. Etest strips were placed on the plates, and the minimal inhibitory concentration (MIC) was read after incubation (48 h at 37 °C). Data were analyzed by a factorial analysis of variance and a 2 × 2 post hoc test (α = .05). RESULTS: C glabrata showed the highest MIC values (P < .001) against caspofungin, itraconazole, and fluconazole. For amphotericin B, the MIC values of C tropicalis and C glabrata (P = .0521) were higher than those of C albicans (P < .001). Itraconazole was the least effective antifungal; 93.3% of the C glabrata isolates, 3.3% of the C albicans, and 1.3% of the C tropicalis were resistant. All microorganisms were susceptible to caspofungin and amphotericin B. CONCLUSIONS: Caspofungin and amphotericin B should be recommended as an effective alternative for the management of oral Candida infections when treatment with topical or other systemic drugs has definitely failed.

Eradication of a Mature Methicillin-Resistant Staphylococcus aureus (MRSA) Biofilm From Acrylic Surfaces

Methicillin-resistant Staphylococcus aureus (MRSA) can grow as structured biofilm in different surfaces, including oral mucosa and denture surfaces. Such biofilms can be released into the oral fluids and aspirated, causing systemic infections such as aspiration pneumonia. This study evaluated the efficacy of two disinfectant solutions and microwave irradiation in disinfecting acrylic specimens contaminated with MRSA biofilm. Thirty-six acrylic specimens were made, sterilized and contaminated with MRSA (107 cfu/mL). After incubation (37 °C/48 h), the specimens were divided into 4 groups: not disinfected (positive control); soaking in 1% sodium hypochlorite for 10 min; soaking in 2% chlorhexidine gluconate for 10 min; and irradiating by microwave for 3 min at 650 W. The viability of cells was evaluated by XTT reduction method. All specimens from the positive control group showed biofilm formation after 48 h incubation. The mean absorbance value of the control specimens was 1.58 (OD at 492 nm). No evidence of biofilm formation was observed on specimens after the disinfection methods. Disinfection by soaking in 1% sodium hypochlorite and 2% chlorhexidine gluconate and irradiating by microwaves resulted in 100% reduction of MRSA biofilm metabolism. The use of chemical solutions and microwave irradiation was shown to be effective for eradicating mature MRSA biofilms on acrylic resin specimens.

Staphylococcus aureus resistente à meticilina (MRSA, do inglês methicillin-resistant Staphylococcus aureus) pode crescer como biofilme estruturado em diferentes superfícies, incluindo mucosa bucal e superfícies de próteses. Estes biofilmes podem se dispersar nos fluidos orais e ser aspirados, causando infecções sistêmicas, como a pneumonia aspirativa. Este estudo avaliou a eficácia de duas soluções desinfetantes e irradiação por microondas na desinfecção de corpos-de-prova acrílicos contaminados com biofilme de MRSA. Trinta e seis espécimes de resina acrílica foram fabricados, esterilizados e contaminados com MRSA (107 ufc/mL). Após a incubação (37 °C/48 h), os espécimes foram divididos em quatro grupos: não desinfetados (controle positivo); imersos em hipoclorito de sódio 1% por 10 min; imersos em gluconato de clorexidina 2% por 10 min e irradiados por microondas durante 3 min a 650 W. A viabilidade das células foi avaliada pelo método de redução de XTT. Todos os espécimes do grupo controle apresentaram formação de biofilme após 48 h de incubação. O valor médio de absorbância destes espécimes foi de 1.58 (OD a 492 nm). Nenhuma evidência de formação de biofilme foi observada em todas as amostras desinfetadas. A desinfecção em hipoclorito de sódio 1%, gluconato de clorexidina 2% e irradiação em microondas resultou em 100% de redução do metabolismo do biofilme de MRSA. O uso de soluções químicas e irradiação em microondas mostrou-se eficaz na eliminação do biofilme maduro de MRSA sobre corpos-de-prova de resina acrílica.

Eradication of a mature methicillin-resistant Staphylococcus aureus (MRSA) biofilm from acrylic surfaces.

Methicillin-resistant Staphylococcus aureus (MRSA) can grow as structured biofilm in different surfaces, including oral mucosa and denture surfaces. Such biofilms can be released into the oral fluids and aspirated, causing systemic infections such as aspiration pneumonia. This study evaluated the efficacy of two disinfectant solutions and microwave irradiation in disinfecting acrylic specimens contaminated with MRSA biofilm. Thirty-six acrylic specimens were made, sterilized and contaminated with MRSA (107 cfu/mL). After incubation (37 °C/48 h), the specimens were divided into 4 groups: not disinfected (positive control); soaking in 1% sodium hypochlorite for 10 min; soaking in 2% chlorhexidine gluconate for 10 min; and irradiating by microwave for 3 min at 650 W. The viability of cells was evaluated by XTT reduction method. All specimens from the positive control group showed biofilm formation after 48 h incubation. The mean absorbance value of the control specimens was 1.58 (OD at 492 nm). No evidence of biofilm formation was observed on specimens after the disinfection methods. Disinfection by soaking in 1% sodium hypochlorite and 2% chlorhexidine gluconate and irradiating by microwaves resulted in 100% reduction of MRSA biofilm metabolism. The use of chemical solutions and microwave irradiation was shown to be effective for eradicating mature MRSA biofilms on acrylic resin specimens.

Effectiveness of mechanical brushing with different denture cleansing agents in reducing in vitro Candida albicans biofilm viability

The adhesion of Candida albicans to surfaces is the prerequisite for occurrence of denture stomatitis, a common disease diagnosed among denture wearers. A routine of denture cleansing is essential to prevent biofilm formation and the onset of this infection. The aim of this study was to investigate the effectiveness of combining brushing and cleansing agents in killing C. albicans biofilm. Disks of acrylic resin were made, sterilized, and inoculated with C. albicans (10(7) cfu/mL). After incubation (37°C/48 h), specimens were randomly assigned to 10 experimental groups (n=9): 5 subjected to brushing with distilled water or cleansing agents - dentifrice slurry, 2% chlorhexidine gluconate (CHX), 1% sodium hypochlorite (NaOCl), and Polident fresh cleanse® (combined method) - and 4 exposed to the cleansing agents without brushing (immersion). Non-cleansed specimens were used as positive controls. The viability of cells was evaluated by XTT reduction method. Results were analyzed by Mann-Whitney and Kruskal-Wallis tests (α=0.05). The combined method was significantly more effective (p<0.0001) in reducing biofilm viability than the immersion. Brushing with CHX and NaOCl resulted in 100% removal of the biofilm. Immersion in the agents reduced significantly (p<0.0001) the biofilm viability, with CHX being the most effective (p<0.0001). The use of the combined method of brushing with cleansing agents is an effective method to reduce C. albicans biofilm, being CHX and NaOCl the most effective solutions.

A adesão de Candida albicans às superfícies é o primeiro passo para o desenvolvimento da estomatite protética, uma infecção frequente diagnosticada entre os usuários de próteses. Uma adequada higienização é essencial para prevenir a formação de biofilme microbiano e o início desta infecção. O objetivo deste estudo foi avaliar a efetividade da escovação com diferentes soluções na eliminação de biofilme de C. albicans. Para isso, discos de resina acrílica foram confeccionados, esterilizados e inoculados com uma suspensão de 10(7) células/mL de C. albicans. Após incubação (37°C/48 h), os espécimes foram aleatoriamente divididos em 10 grupos experimentais (n=9): 5 submetidos à escovação com água ou agentes de limpeza (água destilada, dentifrício, digluconato de clorexidina (CHX) a 2%, hipoclorito de sódio (NaOCl) a 1% e Polident fresh cleanse®) e 4 apenas imersos nos agentes de limpeza. Espécimes não submetidos à higienização foram utilizados como controle positivo. A viabilidade celular foi verificada pelo teste de redução do XTT. Os resultados obtidos foram analisados pelos testes de Mann-Whitney e Kruskal-Wallis (α=0,05). A escovação com todos os agentes de limpeza apresentou redução significativamente superior (p<0,0001) na viabilidade do biofilme quando comparada à exposição dos espécimes às soluções. Escovação com CHX a 2% e NaOCl a 1% resultaram em 100% de inativação do biofilme. A exposição aos agentes de limpeza resultou em redução significativa (p<0,0001) na viabilidade celular, com CHX a 2% sendo o mais efetivo (p<0,0001). A utilização de agentes de limpeza em associação ao método de escovação provou ser efetivo para reduzir biofilme C. albicans, sendo as soluções de CHX e NaOCl as mais efetivas.

Comparison of denture microwave disinfection and conventional antifungal therapy in the treatment of denture stomatitis: a randomized clinical study.

OBJECTIVE: The aim of this study was to compare the effectiveness of denture microwave disinfection and antifungal therapy on treatment of denture stomatitis. STUDY DESIGN: Sixty denture wearers with denture stomatitis (3 groups; n = 20 each), were treated with nystatin or denture microwave disinfection (1 or 3 times/wk) for 14 days. Mycologic samples from palates and dentures were quantified and identified with the use of Chromagar, and clinical photographs of palates were taken. Microbiologic and clinical data were analyzed with the use of a series of statistical tests (α = .05). RESULTS: Both treatments similarly reduced clinical signs of denture stomatitis and growth on palates and dentures at days 14 and 30 (P > .05). At sequential appointments, the predominant species (P < .01) isolated was C. albicans (range 98%-53%), followed by C. glabrata (range 22%-12%) and C. tropicalis (range 25%-7%). CONCLUSIONS: Microwave disinfection, at once per week for 2 treatments, was as effective as topical antifungal therapy for treating denture stomatitis.

Microwave denture disinfection versus nystatin in treating patients with well-controlled type 2 diabetes and denture stomatitis: a randomized clinical trial.

PURPOSE: The aim of this randomized clinical trial was to compare the effectiveness of microwave denture disinfection and nystatin in the treatment of well-controlled type 2 diabetic patients with denture stomatitis in terms of microbiologic and clinical outcomes. MATERIALS AND METHOD: Diabetic patients wearing maxillary complete dentures with denture stomatitis (n = 40) were divided into two groups: NYS (patients treated with topical nystatin 4 times/day for 14 days) and MW (patients who had their dentures microwaved [650 W for 3 minutes] 3 times/week for 14 days). Mycologic samples were taken from the palates and dentures of the patients for quantification and identification of Candida, and standardized photographs of the palates were taken for clinical analysis. Evaluations were repeated at baseline, the end of treatment (day 14), and throughout follow-up (days 30, 60, and 90). Microbiologic data were evaluated by analysis of variance using a random effects statistical model, Tukey post hoc test, and chi-square test (α = .05). Clinical results were analyzed using Mann-Whitney and Fisher exact tests (α = .05). RESULTS: Both treatments were considered successful in reducing the clinical signs of denture stomatitis and significantly reduced the values of colony-forming units/mL from the palates and dentures at days 14 and 30. In addition, 40% of treated patients were cured by the end of treatment. No significant differences in the microbiologic and clinical outcomes were revealed between the two groups (P > .05). C albicans was the most predominant species isolated (P < .01), followed by C tropicalis and C glabrata. CONCLUSION: Denture microwave disinfection was as effective as nystatin for the treatment of diabetic patients with denture stomatitis.

Effectiveness of two disinfectant solutions and microwave irradiation in disinfecting complete dentures contaminated with methicillin-resistant Staphylococcus aureus.

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) biofilm on dentures can be aspirated, thus causing infections such as aspiration pneumonia. The authors evaluated the efficacy of two disinfectant solutions and microwave irradiation in disinfecting complete dentures contaminated with MRSA. METHODS: The authors contaminated 36 simulated complete dentures with MRSA and divided them into four equal groups: a positive control group consisting of dentures that were not disinfected; a group that soaked in 1 percent sodium hypochlorite for 10 minutes; a group that soaked in 2 percent chlorhexidine gluconate for 10 minutes; and a group that underwent microwave irradiation at 650 watts for three minutes. The authors quantified colony counts and evaluated the long-term effectiveness of disinfection. RESULTS: All dentures from the control group showed substantial microbial growth on the plates (6.24 log(10) colony-forming units per milliliter). The authors observed no evidence of microbial growth on plates of any disinfected dentures. After seven days' incubation, the authors observed broth turbidity in all beakers containing the dentures disinfected with 1 percent sodium hypochlorite. CONCLUSIONS: Soaking in chlorhexidine gluconate solution and microwave irradiation resulted in complete disinfection of all dentures contaminated with MRSA in both the short and the long term. Soaking in sodium hypochlorite solution was effective only as a short-term disinfectant. CLINICAL IMPLICATIONS: Microwave irradiation and 2 percent chlorhexidine gluconate may have a disinfective application in dental offices and institutions in which denture wearers are treated, thus improving the longevity and quality of life of patients and reducing the burden of disease caused by MRSA.