Hydrogen peroxide enhances the efficacy of photodynamic therapy against Candida albicans biofilms.

The present study aimed to evaluate the effectiveness of hydrogen peroxide (H2O2) combined with antimicrobial photodynamic therapy (aPDT) on biofilms formed by Candida albicans strains which are either susceptible to or resistant to fluconazole. Biofilms were grown and treated with H2O2, followed by the application of Photodithazine® (P) and red light-emitting diode (LED) (L) either separately or combined (n = 12). After the treatment, biofilms were evaluated by estimating colony-forming unit ml-1, extracellular matrix components [water -soluble and -insoluble polysaccharides, proteins, extracellular DNA (eDNA)], biomass (total and insoluble dry-weight), and protein concentration. Biofilms formed by both strains presented a significant reduction in cell viability, biomass, extracellular matrix components (both types of polysaccharides, eDNA), and proteins (in the soluble and insoluble portion of biofilms) compared to the control. Microscopy images of the biofilms after treatments showed disarticulation of the matrix and scattered fungal cells. The application of H2O2 can disturb the organization of the extracellular matrix, and its association with aPDT potentiated the effect of the treatment.

DNase enhances photodynamic therapy against fluconazole-resistant Candida albicans biofilms.

OBJECTIVE: This study evaluated the effectiveness of DNase I combined with antimicrobial photodynamic therapy, mediated by Photodithazine® and light-emitting diode light, against biofilms formed by a fluconazole-resistant Candida albicans strain (ATCC 96901) and two clinical isolates (R14 and R70). MATERIALS AND METHODS: Biofilms were grown for 48 h and exposed to DNase for 5 min, followed by application of a photosensitizer (P) and light (L), either singly or combined (P+L+, P-L+, P+L-, P-L-, P-L-DNase, P+L+DNase, P+L-DNase, and P-L+DNase; n = 12). Biofilm analysis included quantification of extracellular matrix components (water-soluble and insoluble polysaccharides, proteins and extracellular DNA), and biomass (total and insoluble), as well as the enumeration of colony-forming units. The data were analyzed using three-way analysis of variance with Bonferroni's post hoc test. RESULTS: The DNase treatment combined with aPDT showed a reduction of 1.92, 1.65, and 1.29 log10 of cell viability compared with untreated controls for ATCC 96901, R14, and R70 strains, respectively. It also reduced extracellular matrix contents of water-soluble polysaccharides (36.3%) and extracellular DNA (72.3%), as well as insoluble biomass content (43.3%). CONCLUSION: The three strains showed similar behavior when treated with DNase, and the extracellular matrix components were affected, improving the effectiveness of antimicrobial photodynamic therapy.

Use of Photodynamic Therapy Associated with Antimicrobial Peptides for Bacterial Control: A Systematic Review and Meta-Analysis.

Considering the challenges related to antimicrobial resistance, other strategies for controlling infections have been suggested, such as antimicrobial photodynamic therapy (aPDT) and antimicrobial peptides (AMP). This study aims to perform a systematic review and meta-analysis to obtain evidence on the antimicrobial effectiveness of aPDT associated with AMP and establish in vitro knowledge on this topic for further study designs. The PubMed, Scopus, Web of Science, Science Direct, Scielo, and Cochrane Library databases were searched. Two independent and calibrated researchers (Kappa = 0.88) performed all the systematic steps according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). The odds ratio (OR) was used as the effect measure. The Peto method was used to perform the meta-analysis due to the sparse data. Twenty studies were included in the present review. The result was significant (OR = 0.14/p = 0.0235/I-squared = 0%), showing better outcomes of aPDT associated with peptides than those of aPDT alone for controlling the microbial load. Only 20% of the studies included evaluated this approach in a biofilm culture. Combined treatment with aPDT and AMP highly increased the ability of microbial reduction of Gram-positive and Gram-negative bacteria. However, additional blind studies are required to evaluate the efficacy of this therapy on microbial biofilms.

Lactobacillus casei reduces the extracellular matrix components of fluconazole-susceptible Candida albicans biofilms.

Fluconazole-sensitive (CaS) and -resistant (CaR) C. albicans were grown as single-species and dual-species biofilms with Lactobacillus casei (Lc) and Lactobacillus rhamnosus (Lr). Single-species Lc and Lr were also evaluated. Biofilm analysis included viable plate counts, the extracellular matrix components, biomass, and structural organization. Lc reduced the viability of CaS, water-soluble polysaccharides, and eDNA in CaS + Lc biofilm. Lc biofilm presented more eDNA than CaS. The total biomass of CaS + Lc biofilm was higher than the single-species biofilms. The viability of Lc and Lr was reduced by CaR dual-species biofilms. The total and insoluble biomass in CaS + Lr was higher than in single-species CaS biofilms. Lc hindered the growth of CaS, and their association hampered matrix components linked to the structural integrity of the biofilm. These findings allow understanding of how the implementation of probiotics influences the growth of C. albicans biofilms and thereby helps with the development of novel approaches to control these biofilms.

Photodithazine-mediated antimicrobial photodynamic therapy against fluconazole-resistant Candida albicans in vivo.

This study evaluated the efficacy of antimicrobial Photodynamic Therapy (aPDT) against fluconazole-resistant Candida albicans in a murine model of oral candidosis. Mice were inoculated with two clinical isolates (R10, R15) and one reference strain (ATCC) of resistant C. albicans to produce oral candidosis. After inoculation, aPDT mediated by Photodithazine® (PDZ) and LED light was performed. The use of PDZ or light only was also investigated. Additional animals were treated with Nystatin (NYS). Untreated or healthy mice were also evaluated. Microbiological evaluation was performed by recovering C. albicans from the tongue via colony-forming units. Animals were killed 24 hours after treatments, and the tongues were removed for histological analysis. Data were analyzed by one-way ANOVA and Tukey test (P < .05). The results demonstrated that all strains showed the same behavior after aPDT and NYS treatment. A significant reduction in C. albicans viability was achieved after both treatments for R15 and ATCC. No significant reduction was verified for C. albicans R10 submitted to aPDT or NYS. The histological analysis revealed that aPDT did not cause side effects on tissues. aPDT was effective for inactivation of two fluconazole-resistant C. albicans of the three strains evaluated.

Antimicrobial sonodynamic and photodynamic therapies against Candida albicans.

Candida albicans biofilms exhibit unique characteristics and are highly resistant to antifungal agents. Antimicrobial photodynamic therapy (aPDT) is an alternative treatment limited to treating superficial infections due to the poor light penetration. In this manuscript, the antifungal properties of sonodynamic therapy (SDT) were assessed. SDT uses ultrasound instead of light, enabling the treatment of deeper infections. Planktonic cells and biofilms of C. albicans were treated with aPDT or SDT, in addition to combined aPDT/SDT, with cell survival determined using colony forming units. The total biomass and structural integrity of the biofilms were also investigated. The results demonstrated that while individual aPDT or SDT eradicated suspensions, they had little impact on biofilms. However, combined aPDT/SDT significantly reduced the viability and total biomass of biofilms. Microscopic images revealed that biofilms treated with aPDT/SDT were thinner and comprised mainly of dead cells. These results highlight the potential of combined aPDT/SDT for the inactivation of C. albicans biofilms.

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.

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.

Antimicrobial Photodynamic Therapy Mediated by Curcumin-Loaded Polymeric Nanoparticles in a Murine Model of Oral Candidiasis.

Antimicrobial photodynamic therapy (aPDT) has been proposed as an alternative method for oral candidiasis (OC), while nanocarriers have been used to improve the water solubility of curcumin (CUR). The aim of this study is to encapsulate CUR in polymeric nanoparticles (NPs) and to evaluate its photodynamic effects on a murine model of OC. Anionic and cationic CUR-NP is synthesized using poly-lactic acid and dextran sulfate and then characterized. Female mice are immunosuppressed and inoculated with Candida albicans (Ca) to induce OC. aPDT is performed by applying CUR-NP or free CUR on the dorsum of the tongue, followed by blue light irradiation for five consecutive days. Nystatin is used as positive control. Afterward, Ca are recovered and cultivated. Animals are euthanized for histological, immunohistochemical, and DNA damage evaluation. Encapsulation in NP improves the water solubility of CUR. Nystatin shows the highest reduction of Ca, followed by aPDT mediated by free CUR, which results in immunolabelling of cytokeratins closer to those observed for healthy animals. Anionic CUR-NP does not show antifungal effect, and cationic CUR-NP reduces Ca even in the absence of light. DNA damage is associated with Ca infection. Consecutive aPDT application is a safe treatment for OC.

Virulence factors of fluconazole-susceptible and fluconazole-resistant Candida albicans after antimicrobial photodynamic therapy.

This study evaluated the effects of antimicrobial photodynamic therapy (aPDT) mediated by Photodithazine® (PDZ) and LED light on the virulence factors of fluconazole-susceptible (CaS) and fluconazole-resistant (CaR) Candida albicans. Standardized suspensions of strains were prepared (107), and after 48 h of biofilm formation, these strains were incubated with PDZ (100 mg/L) for 20 min and exposed to LED light (660 nm, 37.5 J/cm2). Additional samples were treated with PDZ or light only, and the control consisted of biofilms that received no treatment. After aPDT, the cells were recovered and the virulence factors were evaluated. To analyze the capacity of adhesion, cells were recovered after aPDT and submitted to the adhesion process in the bottom of a 96-well plate. After this, metabolic activity tests (XTT assay) and cell viability (colony forming units per milliliter, CFU/mL) were applied. To evaluate the biofilm-forming ability after aPDT, the cells recovered were submitted to biofilm formation procedures, and the biofilm formed was evaluated by XTT, CFU/mL, and total biomass (crystal violet) tests. Lastly, the capacity for synthesizing protease and phospholipase enzymes after aPDT was evaluated by fluorimetric tests. Data were analyzed by two- or three-way ANOVA tests (p ≤ 0.05). It was verified that aPDT reduced the viability of both strains, fluconazole-susceptible and fluconazole-resistant C. albicans. It was also observed that the CaR strain had lower susceptibility to the aPDT when compared with the CaS strain. However, regarding the virulence factors evaluated, it was demonstrated that aPDT did not alter the adherence and biofilm formation ability and enzymatic production.

Occlusal Pressure Analysis of Complete Dentures after Microwave Disinfection: A Clinical Study.

PURPOSE: This clinical study evaluated the effect of microwave disinfection protocols on the occlusal pressure pattern of dentures. MATERIALS AND METHODS: Dentures were constructed for 40 patients and evaluated as follows (n = 20). Group 1: Patients had the maxillary dentures submitted to microwave disinfection, once a week, for 4 weeks. Group 2: Patients had the maxillary dentures submitted to microwave disinfection, three times a week, for 4 weeks. Occlusal contacts were recorded on five occasions: 30 days after denture insertion and before first disinfection (baseline or control group); 1 week after disinfection; 2 weeks after disinfection; 3 weeks after disinfection; 4 weeks after disinfection. Occlusal contacts were analyzed by T-Scan III. Intergroup analysis was performed using the Mann-Whitney test and intragroup analysis using the Friedman test with significance of 5%. RESULTS: The results showed no significant difference between groups during the periods. The data on parameters loss of denture adaptation or complaints showed that patients used their dentures regularly for eating and expressed comfort and satisfaction in all experimental periods. The evaluation of functional occlusion revealed that the distribution of the occlusal contacts remained unaltered after disinfection. CONCLUSION: Microwave disinfection protocols as studied in this report did not influence occlusal contacts of the complete dentures.

Cytotoxicity of antimicrobial photodynamic inactivation on epithelial cells when co-cultured with Candida albicans.

This study assessed the cytotoxicity of antimicrobial Photodynamic Inactivation (aPDI), mediated by curcumin, using human keratinocytes co-cultured with Candida albicans. Cells and microorganisms were grown separately for 24 hours and then kept in contact for an additional 24 hours. After this period, aPDI was applied. The conditions tested were: P+L+ (experimental group aPDI); P-L+ (light emitting diode [LED] group); P+L- (curcumin group); and P-L- (cells in co-culture without curcumin nor LED). In addition, keratinocytes and C. albicans were grown separately, were not placed in the co-culture and did not receive aPDI (control group). Cell proliferation was assessed using Alamar Blue, MTT, XTT and CFU tests. Qualitative and quantitative analyses were performed. Analysis of variance (ANOVA) was applied to the survival percentages of cells compared to the control group (considered as 100% viability), complemented by multiple comparisons using Tukey's test. A 5% significance level was adopted. The results of this study showed no interference in the metabolism of the cells in co-culture, since no differences were observed between the control group (cultured cells by themselves) and the P-L- group (co-culture cells without aPDI). The aPDI group reached the highest reduction (p = 0.009), which was equivalent to 1.7 log10 when compared to the control group. The P+L-, P-L+, P-L- and control groups were not statistically different (ρ > 0.05). aPDI inhibited the growth of keratinocytes and C. albicans in all tests, so the therapy was considered slightly (inhibition between 25 and 50% compared to the control group) to moderately (inhibition between 50 and 75% compared to the control group) cytotoxic.

Photodynamic inactivation of a multispecies biofilm using curcumin and LED light.

This study evaluated the potential of curcumin-mediated antimicrobial photodynamic inactivation (API) on multispecies biofilms of Candida albicans, Candida glabrata, and Streptococcus mutans of different ages. Acrylic samples (n = 480) were made with standardized rough surfaces and incubated with bacteria and yeast for 24 or 48 h. API was performed with curcumin (80, 100, 120 µM) and LED light. Additional acrylic samples were treated with curcumin or LED light only. Positive control samples received neither light nor curcumin. After API, colony counts were quantified (CFU/mL), cell metabolism was determined by means of XTT assay, and the total biofilm biomass was evaluated using Crystal Violet (CV) staining assay and images were obtained by confocal laser scanning microscopy (CLSM). The data were analyzed by nonparametric two-way ANOVA and post hoc Tukey tests (α < 0.05). For 24-h biofilm, API resulted in statistically significant difference (ρ < 0.001) of viability of C. albicans compared with control (P-L-) for all Cur concentrations. For 48-h biofilm, API resulted in statistically significant difference (ρ < 0.001) compared with control only when Cur at 120 µM was used. API promoted statistically significant difference (ρ ≤ 0.001) in the viability of S. mutans and C. glabrata for all Cur concentrations in the two biofilm ages. In addition, API produced a statistically significant difference (ρ < 0.001) of metabolic activity and of total biomass (ρ < 0.001) of multispecies biofilms compared with control for all Cur concentrations. It can be concluded that both 24- and 48-h biofilms were susceptible to API mediated by Cur; however, 24-h biofilm was more sensitive than the 48-h biofilm.

Effect of mechanical toothbrushing combined with different denture cleansers in reducing the viability of a multispecies biofilm on acrylic resins.

PURPOSE: To investigate the efficacy of immersion and brushing with different cleansing agents in reducing the viability of multispecies biofilm on acrylic resins. METHODS: Lucitone 550 (L) and Tokuyama Rebase Fast II (T) specimens (10 x 2 mm) were prepared, sterilized, and inoculated with a suspension of Candida albicans, Candida glabrata, and Streptococcus mutans. Specimens were incubated for 48 hours at 37 degrees C for biofilm formation. Then, they were divided into groups (n = 12) and subjected to brushing or immersion for 10 seconds in distilled water (W), 0.2% peracetic acid-Sterilife (Ac), 1% chlorhexidine digluconate (CHX), 1:1 water/dentifrice solution (D), 1% sodiumhypochlorite (NaOCl), and sodium perborate/Corega Tabs (Pb). Viable microorganisms were evaluated by the XTT assay and colony counts (cfu/mL). Data were performed by ANOVA and Tukey test with 5% significance level. RESULTS: The multispecies biofilm on L and T were killed by brushing or immersion in Ac, CHX, and NaOCl for only 10 seconds.

Genotoxic effect of photodynamic therapy mediated by curcumin on Candida albicans.

Photodynamic therapy (PDT) is a promising method for localized and specific inactivation of fungi and bacteria. A nontoxic light-sensitive compound is taken up by cells, which are then exposed selectively to light, which activates toxicity of the compound. We investigated the potential of sublethal PDT using light-sensitive curcumin (CUR) in combination with blue (455 nm) light to promote reactive oxygen species (ROS) formation in the form of singlet oxygen and DNA damage of Candida albicans. Surprisingly, CUR-mediated PDT but also light alone caused significantly longer comet tails, an indication of DNA damage of C. albicans when compared with the negative control. The intracellular ROS production was also significantly higher for the group treated only with light. However, PDT compared to blue light alone significantly slowed DNA repair. Comet tails decreased during 30 min visualized as a 90% reduction in length in the absence of light for cells treated with light alone, while comet tails of cells treated with PDT only diminished in size about 45%. These results indicate that complex mechanisms may result in PDT in a way that should be considered when choosing the photosensitive compound and other aspects of the treatment design.

Photodynamic inactivation of a multispecies biofilm using Photodithazine(®) and LED light after one and three successive applications.

In this investigation, the effectiveness of successive applications of antimicrobial photodynamic inactivation (API) mediated by Photodithazine(®) (PDZ) and LED light was evaluated against a multispecies biofilm formed by Candida albicans, Candida glabrata, and Streptococcus mutans on denture base acrylic resin. Standard cell suspensions (bacteria and yeast) were inoculated on acrylic resin samples, and the biofilm was grown for 48 h (37 °C/75 rpm). API was performed by the administration of PDZ (175 and 200 mg/L) and exposure to 37.5 J/cm(2) of LED light (660 nm). Additional samples were treated with PDZ or LED light only. Untreated control samples were not submitted to light or PDZ. The conditions described were applied once or in three consecutive applications for all groups. Cell viability was determined by colony counts (CFU/mL), metabolic activity, total biomass, and confocal laser scanning microscopy (CLSM). Data were analyzed by a nonparametric two-way ANOVA and Tukey tests (α = 0.05). The results obtained demonstrated a significant effect (p < 0.05) of number of applications and treatment groups for CFU/mL, and S. mutans showed the highest susceptibility to API. The metabolic activity of the multispecies biofilm was significantly reduced (p < 0.05) after API for both numbers of applications, which were also significantly different (p < 0.05) between them. The total biomass of the biofilm was significantly different (p < 0.05) only between groups submitted to one and three API applications. CLSM showed a visual increase of dead cells after API. API-mediated PDZ was effective in reducing the cell viability of multispecies biofilm. Three consecutive applications of API were more effective for reducing the cell viability and the total biomass of multispecies biofilm.

Susceptibility of multispecies biofilm to photodynamic therapy using Photodithazine®.

This in vitro study evaluated the effect of photodynamic therapy (PDT) on the multispecies biofilm of Candida albicans, Candida glabrata, and Streptococcus mutans. Standardized fungal and bacterial suspensions were cultivated appropriately for each species and inoculated in 96-well microtiter plates for mix-biofilm formation. After 48 h of incubation, the biofilms were submitted to PDT (P + L+) using Photodithazine® (PDZ) at 100, 150, 175, 200, or 250 mg/mL for 20 min and 37.5 J/cm(2) of light-emitting diode (LED) (660 nm). Additional samples were treated only with PDZ (P + L-) or LED (P-L+), or neither (control, P-L-). Afterwards, the biofilms were evaluated by quantification of colonies (CFU/mL), metabolic activity (XTT reduction assay), total biomass (crystal violet staining), and confocal scanning laser microscopy (CSLM). Data were analyzed by one-way ANOVA and Tukey tests (p < 0.05). Compared with the control, PDT promoted a significant reduction in colonies viability of the three species evaluated with 175 and 200 mg/mL of PDZ. PDT also significantly reduced the metabolic activity of the biofilms compared with the control, despite the PDZ concentration. However, no significant difference was found in the total biomass of samples submitted or not to PDT. For all analysis, no significant difference was verified among P-L-, P + L-, and P-L+. CSLM showed a visual increase of dead cells after PDT. PDT-mediated PDZ was effective in reducing the cell viability of multispecies biofilm.

Antimicrobial photodynamic therapy against pathogenic bacterial suspensions and biofilms using chloro-aluminum phthalocyanine encapsulated in nanoemulsions.

Antimicrobial photodynamic therapy represents an alternative method of killing resistant pathogens. Efforts have been made to develop delivery systems for hydrophobic drugs to improve the photokilling. This study evaluated the photodynamic effect of chloro-aluminum phthalocyanine (ClAlPc) encapsulated in nanoemulsions (NE) on methicillin-susceptible and methicillin-resistant Staphylococcus aureus suspensions and biofilms. Suspensions and biofilms were treated with different delivery systems containing ClAlPc. After the pre-incubation period, the drug was washed-out and irradiation was performed with LED source (660 ± 3 nm). Negative control samples were not exposed to ClAlPc or light. For the suspensions, colonies were counted (colony-forming units per milliliter (CFU/mL)). The metabolic activity of S. aureus suspensions and biofilms were evaluated by the XTT assay. The efficiency was dependent on the delivery system, superficial load and light dose. Cationic NE-ClAlPc and free-ClAlPc caused photokilling of the both strains of S. aureus. For biofilms, cationic NE-ClAlPc reduced cell metabolism by 80 and 73% of susceptible and resistant strains, respectively. Although anionic NE-ClAlPc caused a significant CFU/ml reduction for MSSA and MRSA, it was not capable of reducing MRSA biofilm metabolism. This therapy may represent an alternative treatment for eradicating resistant strains.

Effects of short-term immersion and brushing with different denture cleansers on the roughness, hardness, and color of two types of acrylic resin.

PURPOSE: To investigate the cumulative effects of brushing (B) or immersion (I), using different cleansing agents, on the surface roughness, hardness and color stability of a heat-polymerized denture resin, Lucitone 550 (L), and a hard chairside reline resin, Tokuyama Rebase Fast II (T). METHODS: A total of 316 specimens (10 x 2 mm) were fabricated. The specimens (n = 9) were divided into brushing or immersion groups according to the following agents: dentifrice/distilled water (D), 1% sodium hypochlorite (NaOCl), Corega Tabs (Pb), 1% chlorhexidine gluconate (Chx), and 0.2% peracetic acid (Ac). Brushing and immersion were tested independently. Assays were performed after 1, 3, 21, 45 and 90 brushing cycles or immersion of 10 seconds each. Data were evaluated statistically by repeated measures ANOVA. Tukey's honestly significant difference (HSD) post-hoc test was used to determine differences between means (α = 0.05). RESULTS: For L there was no statistically significant difference in roughness, except a significant decrease in roughness by brushing with D. T showed a significant effect on the roughness after 90 immersions with Ac. Hardness values decreased for L when specimens were immersed or brushed in NaOCl and Pb. The hardness of T decreased with increases in the repetitions (immersion or brushing), regardless of the cleaning method. Values of color stability for L resin showed significant color change after brushing with and immersion in Ac and Pb. Brushing with D exhibited a higher incidence of color change. For T there were no significant differences between cleaning agents and repetitions in immersion. A color change was noted after three brushings with the Ac, Chx, and D. Brushing with dentifrice decreased roughness of L. Immersion in or brushing with NaOCl and Pb decreased the hardness of L. For T, hardness decreased with increases in immersions or brushing. Color changes after the immersion in or brushing with cleaning agents were clinically acceptable according to National Bureau of Standards parameters for both resins.

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.