Cryptocarya moschata extract decreases single and mixed biofilms on acrylic resins.

OBJECTIVE: This study proposed to assess the effect of Cryptocarya moschata extract on single and mixed biofilms formed on denture base and reline acrylic resin. MATERIALS AND METHODS: Single and mixed biofilms of Candida albicans and Streptococcus mutans were formed on the samples and treated with C. moschata extract; Nystatin solution at 100,000 IU/mL or Penicillin antibiotic solution at 100,000 IU/mL; or PBS solution. Antimicrobial activity was analyzed by counting colony-forming units, metabolism assay, assessment of protein components of the biofilm matrix, and of cell viability using confocal laser scanning microscopy (CLSM). Data were submitted to ANOVA and Tukey's post-test (α = 0.05). RESULTS: Cryptocarya moschata extract reduced cell viability of C. albicans and S. mutans single and mixed biofilms formed on samples. For all types of biofilms in the C. moschata group, there was a log reduction of the biofilm, proven by the Alamar Blue assay. Analyzing the extracellular matrix protein components, groups treated with the extract exhibited a lower level of fluorescence compared to the PBS groups. Reduction in thickness biofilm and viable cells was perceptible in the C. moschata group when assessing through CLSM. CONCLUSION: Cryptocarya moschata extract reduced the single and mixed biofilms of C. albicans and S. mutans on acrylic resins.

Candida species as potential risk factors for oral squamous cell carcinoma: Systematic review and meta-analysis.

Oral squamous cell carcinoma (OSCC) is considered a multifactorial disease and has been associated with microbial infections, although the association with Candida spp. is still controversial. This systematic review focused on clinical trials which evaluated the relation between oral Candida spp colonization and OSCC. PubMed; Scopus; Embase; Web of Science and Scientific Direct were assessed. Independent reviewers conducted the diagram steps. For data extraction the PRISMA protocol was followed. The quality analysis of case-control studies was performed based on the Newcastle-Ottawa scale. Meta-analysis was performed to evaluate the frequency of Candida spp and the levels of microbial acetaldehyde production (MAP) being odds ratio (OR) the effect-measure applied. Eight and six studies were included in the qualitative analysis and meta-analysis, respectively. It was noted that there was a significantly higher frequency of Candida species (p = 0.0003/OR = 9.50) in patients diagnosed with OSCC than healthy patients, especially Candida krusei (p = 0.0167/OR=4.62). Candida spp., from oral cancer patients demonstrated significantly greater biofilm, biofilm metabolic activity, phospholipase, proteinase activity and a higher production of MAP (p = 0.0111/OR = 2.67). Candida species may have a potential role in OSCC development. Further studies should be conducted to elucidate the mechanism of action of Candida spp and others risk factors in the development of OSCC.

Do denture cleansers influence the surface roughness and adhesion and biofilm formation of Candida albicans on acrylic resin? Systematic review and meta-analysis.

PURPOSE: To evaluate the influence of denture cleansers on the surface roughness, Candida albicans adhesion, and biofilm formation on denture base acrylic resins. STUDY SELECTION: Electronic databases and gray literature were searched using an individual search strategy. In vitro studies that evaluated the effects of immersion in denture cleansers on the surface roughness (µm) and antimicrobial activity (CFU/mL) on samples of heat-polymerized denture base acrylic resins were included. RESULTS: After screening, 17 studies were included, and a qualitative synthesis was performed. After assessing the risk of bias, only nine studies were included in the meta-analysis. The meta-analysis results showed that the evaluated solutions (0.5% sodium hypochlorite, 1% sodium hypochlorite, alkaline peroxide, and natural substances) did not influence the roughness of the acrylic resin. However, in the qualitative analysis, it was not possible to confirm an association between roughness and C. albicans adhesion and biofilm formation on the acrylic resin samples. CONCLUSION: Denture cleansers did not affect the surface roughness of denture base acrylic resins.

In vivo antifungal activity and biocompatibility of Cryptocarya moschata.

The objective of this study was evaluate, in vivo model, the antifungal activity of Cryptocarya moschata extract against Candida albicans and its biocompatibility. The animals (N = 50) were divided into groups (n = 5): CI/CG: candidiasis was induced and treated with C. moschata extract (0.045 g/mL); CI/NG: candidiasis was induced and treated with nystatin; CI/NT: candidiasis was induced and no treated; CI/CG-2: candidiasis was induced and treated with C. moschata extract (0.045 g/mL), reapplied after 24 h; CI/NG-2: candidiasis was induced and treated with nystatin, reapplied after 24 h; NCI/NT: candidiasis was not induced and no treated; NCI/CG: candidiasis was not induced and treated with C. moschata extract (0.045 g/mL); NCI/NG: candidiasis was not induced treated with nystatin; NCI/CG-2: candidiasis was not induced and treated with C. moschata extract (0.045 g/mL), reapplied after 24 h; NCI/NG-2: candidiasis was not induced and treated with nystatin, reapplied after 24 h. The fungi present in the lingual dorsum of mice were collected and analyzed by the count of colony-forming units. In addition, histological analysis was performed. Histologically, there was no cell damage in the mice's tongue, and there was a decrease in Candida biofilm, similar to the use of nystatin. It was concluded that the C. moschata extract was effective against C. albicans and was biocompatible.

Antimicrobial efficacy and biocompatibility of extracts from Cryptocarya species.

This study evaluated the efficacy of Cryptocarya spp extracts on biofilm of Candida albicans and its biocompatibility. Mature biofilm of C. albicans was formed on denture base acrylic resin samples and the fungicidal effect of the extracts was evaluated by Alamar Blue® assay, counting colony-forming units (CFU/mL) and confocal laser scanning microscopy (CLSM). Cytotoxicity of extracts from Cryptocarya species was evaluated by AlamarBlue® assay, using normal oral keratinocytes (NOK) cells. In additional, Analysis of plant extracts by ultra-high-performance liquid chromatography-diode array detector-tandem mass spectrometry (UPLC-DAD-MS) was performed. The results showed significant reduction in the cellular metabolism and in the number of CFU/mL of C. albicans (p<0.05). The concentration of 0.045 g/mL completely inhibited the number of CFU/mL. Regarding cytotoxicity, all extracts decreased cell viability compared to the control group. CLSM analysis showed predominance of live cells, but with a great difference between the groups. Antimicrobial activity of extracts from Cryptocarya on C. albicans biofilm was confirmed. However, all extracts showed toxicity on NOK cells.

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.

Statherin-derived peptides as antifungal strategy against Candida albicans.

OBJECTIVE: The aim of this in vitro study was to evaluate the effect of statherin and its naturally occurring peptides (DR9-2, DR9, GE-12, IT-32, GQ-19, IP-18) on Candida albicans metabolism and biofilm development. DESIGN: After the killing assay, a peptide pellicle was formed on the bottom of a polystyrene plate at the IC50 of each peptide. Over the peptide pellicle, Candida albicans biofilm (48 h) was grown. The peptides antimicrobial activity after the peptides treatment was evaluated by alamarBlue, total biofilm biomass and colony forming units (CFU) counting. RESULTS: The pellicle with statherin and the peptides (DR9-2, DR9, GE-12, IP-18, GQ-19) was able to reduce he viability of Candida albicans compared to the negative control. They also decreased cell proliferation by 20 % and total biomass. IT-32 showed the highest reduction in cell proliferation and biomass, which was similar to the positive control, histatin 5. CONCLUSIONS: These results suggest that the naturally occuring peptides from statherin are able to decrease Candida albicans colonization and biofilm proliferation.

Effectiveness of Disinfectant Liquid Soaps in the Reduction of Candida spp Present in Complete Dentures: A Crossover Randomized Clinical Trial.

PURPOSE: To evaluate the effectiveness of liquid disinfectant soaps for the reduction of microorganisms present on maxillary complete dentures. MATERIALS AND METHODS: The selected patients (N = 28) were randomly divided into four groups (n = 7), and each group underwent all four disinfection treatments in a different order. The disinfection treatments evaluated were: 0.5% sodium hypochlorite (positive control); Dettol liquid soap; Lifebuoy liquid soap; and phosphate-buffered saline solution (negative control). The patients were instructed to immerse their maxillary dentures in the disinfectant solution for 8 hours (overnight) for 7 days, with the solutions in a randomized sequence with a washout period of 1 week between solutions. Biofilm samples of the dental prostheses were obtained before and after each treatment with a sterile swab, and the microbiologic material was diluted and plated in selective media for Candida spp. Colony-forming unit count (CFU/mL) was performed in each group. One-way ANOVA with Welch correction was used for analysis, with Games-Howell post hoc test with a significance level of .05. RESULTS: A 3-log reduction in microorganisms was considered effective compared to baseline. The highest incidence observed was for Candida albicans, which presented with a frequency between 66% and 92%, followed by C tropicalis, with a frequency between 7% and 33%, in all experimental groups. Sodium hypochlorite was able to reduce more than 3 log10 of microorganisms for all patients, showing high antifungal effectiveness for both C albicans and C tropicalis species. Regarding the experimental groups, both liquid soaps (Dettol and Lifebuoy) were effective in reducing the two types of microorganisms. CONCLUSION: Liquid soaps were effective at reducing biofilm and may be an alternative for disinfection of removable partial dentures or complete dentures.

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.

Effect of the aging of titanium and zirconia abutment surfaces on the viability, adhesion, and proliferation of cells and the adhesion of microorganisms.

STATEMENT OF PROBLEM: The longevity of dental implants depends on the maintenance of peri-implant tissue and absence of inflammation. How the physical-chemical properties intrinsic to each material over time can affect adhesion, given constant cell turnover and biofilm development, remains unclear. PURPOSE: The purpose of this in vitro study was to evaluate the influence of aging on the viability, adhesion, and proliferation of normal oral keratinocytes (Nok-si) and on the multispecies biofilm formation of Fusobacterium nucleatum (F. nucleatum), Porphyromonas gingivalis (P. gingivalis), and Streptococcus sanguinis (S. sanguinis). MATERIAL AND METHODS: Zirconia (ZrO2) and titanium (Ti) disks were analyzed by surface roughness, water contact angle, and X-ray diffraction before and after aging in an autoclave. The Nok-si cell viability was evaluated by using a 3-(4.5-dimethylthiazole-2-yl)2.5-diphenyl tetrazolium bromide assay (MTT), morphology by scanning electron microscopy (SEM), and proliferation and adhesion by using a confocal microscope. Multispecies biofilms were analyzed quantitatively by colony-forming units per milliliter (CFU/mL) and qualitatively by SEM. RESULTS: For Ti, the aging process affected the roughness and wettability. However, for ZrO2, the aging did not affect roughness but did affect wettability and the ratio of the tetragonal to monoclinic phase (P<.05). A significant difference was found in the bacterial growth for Ti (nonaged and aged) in relation to the control, and no differences were found in Ti before and after aging; however, ZrO2 had increased growth of microorganisms after aging. For ZrO2, a statistically significant difference was found between aged ZrO2 and the control (P<.001). CONCLUSIONS: The results indicate that, after the aging, Ti showed better cell adhesion and proliferation and lower biofilm adhesion than zirconia.

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.

Properties of an acrylic resin after immersion in antiseptic soaps: Low-cost, easy-access procedure for the prevention of denture stomatitis.

Denture stomatitis triggered by Candida species requires better preventive measures. This study evaluated the physical and biological properties of a denture base acrylic resin after immersion in antiseptic soaps. Acrylic resin specimens were prepared and stored in distinct solutions for 0, 7, 14, 21, and 28 days. The solutions were as follows: DW: distilled water at 37°C (control group); DS: cycles of daily immersion in Dettol soap for 8 hours at room temperature, followed by immersion in distilled water for 16 hours at 37°C; PS: cycles of daily immersion in Protex soap, as described for the previous group; LS: cycles of daily immersion in Lifebuoy soap, as described for the DS group. The parameters evaluated at each time point were the following: biofilm formation capacity by Candida albicans and reduction of preformed fungal biofilms, cytotoxicity, surface roughness, hardness, and color change. For the fungal adhesion phase, the type of soap had a statistically significant effect (p = 0.0292), but after 24 hours, no differences were found between solutions or between storage times. Regarding the efficacy of biofilm reduction, there was a significant difference when the groups were compared to each other (p = 0.014). Dettol and Lifebuoy eliminated the preformed biofilm on the specimens. Moreover, all the soaps were classified as non-cytotoxic (on HaCaT cell line) because there was no difference in cell viability between the different groups, except after 21 days, when a decrease in cell viability occurred, regardless of the type of soap. Regarding the roughness, there was no statistically significant difference (p > 0.05) between the groups. Lifebuoy decreased resin hardness regardless of storage time (p = 0.003). After 21 and 28 days of storage, there was an increase in hardness value, regardless of the type of soap. The specimens' color, according to the National Bureau of Standards values, ranged from 0.27 to 0.58 (i.e., imperceptible or mild color changes). In general, the disinfectant soaps were not able to prevent biofilm formation, but all the soaps were effective in reducing the preformed biofilm. In addition, all soaps were non-cytotoxic and did not change surface roughness, hardness (except Lifebuoy), and color (except Lifebuoy). Therefore, immersion in two antiseptic soaps (Protex and Dettol) may be a cheap and easy procedure for preventing denture stomatitis.

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.

Effect of surface characteristics of soft liners and tissue conditioners and saliva on the adhesion and biofilm formation.

PURPOSE: To investigate the influence of surface characteristics and saliva on the adhesion and biofilm formation of Candida glabrata and methicillin-resistant Staphylococcus aureus (MRSA) to soft liners and tissue conditioners. METHODS: For each material (Ufi Gel P - UG; Sofreliner S - SS; Trusoft - TR; Coe Comfort - CC; Softone - ST), specimens were prepared and roughness (Ra), hydrophobicity (water contact angles-WCA) and surface free energy (SFE) were measured. Surface morphology was also analyzed using scanning electron microscopy (SEM). Specimens were incubated in C. glabrata or MRSA suspensions for 90 minutes (adhesion) or 48 hours (biofilm). The absorbance (AB) was measured by XTT assay. Experiments were performed using specimens that were either uncoated or had been coated with saliva. Data were analyzed using one- or two-way ANOVAs, followed by Tukey's test (α= 0.05). RESULTS: TR exhibited the highest Ra and UG the lowest. SEM images also showed that UG and SS had smooth surfaces, while TR presented several irregularities and pores. In the absence of saliva, UG and SS presented higher WCA and lower SFE than the other materials. XTT results showed that, in the C. glabrata adhesion assay, the AB value was higher for TR followed by UG > CC> SS> ST. For the biofilm formation of C. glabrata, AB values were in the following order TR > CC = UG > ST = SS. In the adhesion assay, AB values obtained for MRSA were TR > UG = CC > ST > SS and for the biofilm formation were TR > ST > CC > UG > SS. Saliva decreased the WCA and increased the SFE for all materials. In general, the presence of saliva decreased the adhesion and biofilm formation of both microorganisms to the acrylic-based material (TR) and tissue conditioners (CC and ST), and increased for the silicone-based soft liners (UH and SS). Surface characteristics and the influence of saliva varied among materials. Roughness seemed to favor C. glabrata and MRSA adhesion and biofilm formation. CLINICAL SIGNIFICANCE: The presence of microorganisms on denture liners can irritate the oral tissues and contribute to systemic diseases. Colonization with more tolerant microorganisms such as C. glabrata and MRSA may expose patients to a greater risk of infection, mainly in immunocompromised hosts, such as aged individuals after treatment of oral cancer. For this, it is important to investigate the surface characteristics of soft liners and tissue conditioners, as well as saliva, and their influence on the adhesion and biofilm formation of C. glabrata and methicillin-resistant Staphylococcus aureus.

pH-responsive poly(aspartic acid) hydrogel-coated magnetite nanoparticles for biomedical applications.

A novel multifunctional nanosystem formed by magnetite nanoparticles coated with pH-responsive poly(aspartic acid) hydrogel was developed. Magnetite nanoparticles (Fe3O4) have been intensively investigated for biomedical applications due to their magnetic properties and dimensions similar to the biostructures. Poly(aspartic acid) is a water-soluble, biodegradable and biocompatible polymer, which features makes it a potential candidate for biomedical applications. The nanoparticles surface modification was carried out by crosslinking polysuccinimide on the magnetite nanoparticles surface and hydrolyzing the succinimide units in mild alkaline medium to obtain the magnetic poly(aspartic acid) hydrogel. The surface modification in each step was confirmed by DRIFTS, TEM and zeta potential measurements. The hydrodynamic diameter of the nanosystems decreases as the pH value decreases. The nanosystems showed high colloidal stability in water and no cytotoxicity was detected, which make these nanosystems suitable for biomedical applications.

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.

Photoinactivation of single and mixed biofilms of Candida albicans and non-albicans Candida species using Photodythazine® [corrected].

This study evaluated the effectiveness of antimicrobial photodynamic therapy (aPDT) mediated by Photodithazine® (PDZ) formulated in hydrogel, in the inactivation of mono and duo-species biofilms of Candida albicans, Candida glabrata and Candida tropicalis. Standardized suspensions of each strain were prepared and after biofilm formation, mono-species were treated with 150 and 175mg/L of PDZ for 20min (pre-irradiation time), and exposed to LED light at a dose of 37.5J/cm2 (660nm). The duo-species biofilms (C. albicans+C. glabrata and C. albicans+C. tropicalis) were treated with 150mg/L of PDZ and light. Additional samples were treated with PDZ or light only, and the control did not receive any treatment. Next, microbiological evaluation was performed by spreading the cells on Sabouraud Dextrose Agar and CHROMagar Candida for colony forming units (CFU/mL). Moreover, the total biomass of biofilm was verified using the crystal violet staining assay (CV). The data were submitted to ANOVA and Tukey post-hoc (α=0.05). The use of PDZ 150mg/L promoted a reduction of 1.0, 1.2, 1.5 log10 in the viability of C. glabrata, C. albicans and C. tropicalis, respectively. The same concentration reduced in 1.0 log10 the viability of each species grown as duo-species biofilms. The crystal violet assay showed that the use of 150mg/L reduced 24.4%, 39.2% and 43.7% of the total biomass of C. albicans, C. tropicalis and C. glabrata, respectively. aPDT did not reduce the total biomass to the duo-species biofilms. Thus, PDZ-mediated aPDT was more effective in the inactivation of mono-species biofilms of Candida spp. compared with duo-species biofilm.

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.