Lívia Nordi Dovigo

Investigation of the photodynamic effects of curcumin against Candida albicans.

This study describes the association of curcumin with light emitting diode (LED) for the inactivation of Candida albicans. Suspensions of Candida were treated with nine curcumin concentrations and exposed to LED at different fluences. The protocol that showed the best outcomes for Candida inactivation was selected to evaluate the effect of the preirradiation time (PIT) on photodynamic therapy (PDT) effectiveness, the uptake of curcumin by C. albicans cells and the possible involvement of singlet oxygen in the photodynamic action. Curcumin‐mediated PDT was also assessed against biofilms. In addition to the microbiological experiments, similar protocols were tested on a macrophage cell line and the effect was evaluated by Methyltetrazolium assay (MTT) and SEM analysis. The optical properties of curcumin were investigated as a function of illumination fluence. When compared with the control group, a statistically significant reduction in C. albicans viability was observed after PDT (P < 0.05), for both planktonic and biofilm cultures. Photodynamic effect was greatly increased with the presence of curcumin in the surrounding media and the PIT of 20 min improved PDT effectiveness against biofilms. Although PDT was phototoxic to macrophages, the therapy was more effective in inactivating the yeast cell than the defense cell. The spectral changes showed a high photobleaching rate of curcumin.

Susceptibility of Candida albicans to photodynamic therapy in a murine model of oral candidosis

OBJECTIVE In vivo studies of antimicrobial PDT in animal models of oral candidosis are scarce and the association of porphyrin and LED light has not been evaluated for in vivo photoinactivation of Candida. In this study the effectiveness of photodynamic therapy (PDT) on the inactivation of Candida albicans in vivo was evaluated. STUDY DESIGN Seventy-one 6-week-old female Swiss mice were immunosuppressed, provided tetracycline to their drinking water, then orally swabbed with a suspension of C. albicans (10(7) CFU/mL). Four days after oral inoculation, PDT was performed on the dorsum of the tongue after topical administration of Photogem at 400, 500, or 1000 mg/L and followed 30 minutes later by illumination with LED light (305 J/cm(2)) at 455 or 630 nm (n = 5 each). After swabbing to recover yeast from the tongue, the number of surviving yeast cells was determined (CFU/mL) and analyzed by ANOVA and Holm-Sidak tests (P < .05). Animals were humanely killed, and the tongues surgically removed and processed for histological evaluation of presence of yeast and inflammatory reaction. RESULTS PDT resulted in a significant reduction in C. albicans recovered from the tongue (P < .001) when compared with mice from the positive control group. There was no difference between the concentrations of Photogem and LED light wavelengths used. Histological evaluation of the tongue revealed that PDT causes no significant adverse effects to the local mucosa. CONCLUSION PDT promoted significant reduction in the viability of C. albicans biofilm without harming the tongue tissue.

Fungicidal effect of photodynamic therapy against fluconazole-resistant Candida albicans and Candida glabrata

Although photodynamic therapy (PDT) has shown great promise for the inactivation of Candida species, its effectiveness against azole‐resistant pathogens remains poorly documented. This in vitro study describes the association of Photogem® (Photogem, Moscow, Russia) with LED (light emitting diode) light for the photoinactivation of fluconazole‐resistant (FR) and American Type Culture Collection (ATCC) strains of Candida albicans and Candida glabrata. Suspensions of each Candida strain were treated with five Photogem® concentrations and exposed to four LED light fluences (14, 24, 34 or 50 min of illumination). After incubation (48 h at 37 °C), colonies were counted (CFU ml−1). Single‐species biofilms were generated on cellulose membrane filters, treated with 25.0 mg l−1 of Photogem® and illuminated at 37.5 J cm−2. The biofilms were then disrupted and the viable yeast cells present were determined. Planktonic suspensions of FR strains were effectively killed after PDT. It was observed that the fungicidal effect of PDT was strain‐dependent. Significant decreases in biofilm viability were observed for three strains of C. albicans and for two strains of C. glabrata. The results of this investigation demonstrated that although PDT was effective against Candida species, fluconazole‐resistant strains showed reduced sensitivity to PDT. Moreover, single‐species biofilms were less susceptible to PDT than their planktonic counterparts.

Susceptibility of clinical isolates of Candida to photodynamic effects of curcumin

The resistance of Candida species to antifungals represents a major challenge for therapeutic and prophylactic strategies. This study evaluated photodynamic therapy (PDT) mediated by Curcumin (CUR) against clinical isolates of C. albicans, C. tropicalis, and C. glabrata, both in planktonic and biofilm forms.

Denture disinfection by microwave irradiation: A randomized clinical study

OBJECTIVE This study evaluated the clinical effectiveness of two exposure times of microwave irradiation on the disinfection of complete dentures. METHODS Biofilm samples were collected from dentures of 30 patients, who were randomly divided into two experimental groups of 15 subjects each: Group 1-patients had their maxillary denture microwaved for 3 min (650W); Group 2-patients had their maxillary denture microwaved for 2 min (650W). Denture biofilm samples were taken with swabs, before (left side surfaces) and after (right side surfaces) microwave irradiation. All microbial material was plated on selective media for Candida spp., Staphylococcus spp., mutans streptococci and a non-selective media. After incubation (48 h/37 degrees C), the number of colony-forming units (cfu/mL) was counted. Microorganisms which grew on selective media were identified using biochemical methods. The data were statistically analyzed by Kruskal-Wallis test, followed by Dunns post-test (alpha=0.05). RESULTS Microwave irradiation for 3 min (Group 1) resulted in sterilization of all dentures evaluated. After microwave irradiation for 2 min (Group 2), a significant decrease in Candida spp. (P=0.0062), Staphylococcus spp. (P=0.0178), mutans streptococci (P=0.0047) and non-identified species (P<0.0001) was achieved in comparison with the cfu/mL obtained before irradiation. The colonies grown after 2 min of microwave irradiation were identified as Candida albicans, non-aureus Staphylococci and Streptococcus mutans. CONCLUSION Microwave irradiation for 3 min may be a potential treatment to prevent cross-contamination.

Curcumin-mediated photodynamic inactivation of Candida albicans in a murine model of oral candidiasis

In vitro investigations of curcumin-mediated photodynamic therapy (PDT) are encouraging, but there is a lack of reliable in vivo evidence of its efficacy. This study describes the photoinactivation of Candida albicans in a murine model of oral candidiasis, using curcumin as a photosensitizer. Forty immunosuppressed mice were orally inoculated with C. albicans and after five days, they received topical curcumin (20, 40 and 80 μM) and illumination with LED light. The use of curcumin or light alone were also investigated. Positive control animals did not receive any treatment and negative control animals were not inoculated with C. albicans. The number of surviving yeast cells was determined and analyzed by ANOVA and Tukeys post-hoc test (α = 0.05). Histological evaluation of the presence of yeast and inflammatory reaction was also conducted. All exposures to curcumin with LED light caused a significant reduction in C. albicans viability after PDT, but the use of 80 μM curcumin associated with light was able to induce the highest log10 reduction in colony counts (4 logs). It was concluded that curcumin-mediated PDT proved to be effective for in vivo inactivation of C. albicans without harming the host tissue of mice.

Effect of different pre-irradiation times on curcumin-mediated photodynamic therapy against planktonic cultures and biofilms of Candida spp

OBJECTIVES The aim of this study was to evaluate the effects of pre-irradiation time (PIT) on curcumin (Cur)-mediated photodynamic therapy (PDT) against planktonic and biofilm cultures of reference strains of Candida albicans, Candida glabrata and Candida dubliniensis. MATERIALS AND METHODS Suspensions and biofilms of Candida species were maintained in contact with different concentrations of Cur for time intervals of 1, 5, 10 and 20min before irradiation and LED (light emitting diode) activation. Additional samples were treated only with Cur, without illumination, or only with light, without Cur. Control samples received neither light nor Cur. After PDT, suspensions were plated on Sabouraud Dextrose Agar, while biofilm results were obtained using the XTT-salt reduction method. Confocal Laser Scanning Microscopy (CLSM) observations were performed to supply a better understanding of Cur penetration through the biofilms after 5 and 20min of contact with the cultures. RESULTS Different PITs showed no statistical differences in Cur-mediated PDT of Candida spp. cell suspensions. There was complete inactivation of the three Candida species with the association of 20.0μM Cur after 5, 10 and 20min of PIT. Biofilm cultures showed significant reduction in cell viability after PDT. In general, the three Candida species evaluated in this study suffered higher reductions in cell viability with the association of 40.0μM Cur and 20min of PIT. Additionally, CLSM observations showed different intensities of fluorescence emissions after 5 and 20min of incubation. CONCLUSION Photoinactivation of planktonic cultures was not PIT-dependent. PIT-dependence of the biofilm cultures differed among the species evaluated. Also, CLSM observations confirmed the need of higher time intervals for the Cur to penetrate biofilm structures.

Microwave disinfection of complete dentures contaminated in vitro with selected bacteria.

PURPOSE This study evaluated the effectiveness of microwave irradiation for disinfection of simulated complete dentures. MATERIALS AND METHODS Seventy dentures were fabricated in a standardized procedure, subjected to ethylene oxide sterilization, individually inoculated (10(7) cfu/mL) with Staphylococcus aureus (n = 20), Pseudomona aeruginosa (n = 20), and Bacillus subtilis (n = 30) and incubated for 24 hours at 37 degrees C. After that, 40 dentures were selected for microwaving. For each microorganism, 10 dentures were submitted to microwave irradiation at 650 W for 3 minutes. In addition, 10 dentures contaminated with B. subtilis were irradiated for 5 minutes. Thirty non-microwaved dentures (n = 10 for each bacteria) were used as positive controls. Replicate aliquots (25 microL) of suspensions were plated at dilutions of 10(-3) to 10(-6) on plates of selective media appropriate for each organism. After incubation (37 degrees C for 48 hours), colonies were counted (cfu/mL). TSB beakers with the microwaved dentures were incubated at 37 degrees C for a further 7 days to verify long-term disinfection. The data were statistically analyzed by the Kruskal-Wallis test (alpha= 0.05). RESULTS No evidence of growth was observed at 48 hours for S. aureus and P. aeruginosa on plates, and no turbidity was visible in the TSB beakers of these specimens after 7 days of incubation. Dentures contaminated with B. subtilis and irradiated for 3 minutes produced microbial growth on six plates and turbidity on all TSB beakers. Microwaving for 5 minutes resulted in survival of B. subtilis in two plates and two beakers. CONCLUSION Microwave irradiation for 3 minutes at 650 W produced sterilization of complete dentures contaminated with S. aureus and P. aeruginosa. Dentures contaminated with B. subtilis were disinfected by microwave irradiation after 3 and 5 minutes at 650 W.

Effectiveness of Photodynamic Therapy for the Inactivation of Candida spp. on Dentures: In Vitro Study

OBJECTIVE This in vitro study evaluated the effectiveness of photodynamic therapy (PDT) for the inactivation of different species of Candida on maxillary complete dentures. BACKGROUND DATA The treatment of denture stomatitis requires the inactivation of Candida spp. on dentures. PDT has been reported as an effective method for Candida inactivation. METHODS Reference strains of C. albicans, C. glabrata, C. tropicalis, C. dubliniensis and C. krusei were tested. Thirty-four dentures were fabricated in a standardized procedure and subjected to ethylene oxide sterilization. The dentures were individually inoculated with one of the strains and incubated at 37°C for 24 h. Dentures submitted to PDT (P+L+) were individually sprayed with 50 mg/L of Photogem(®) (PS) and, after 30 min, illuminated by LED light for 26 min (37.5 J/cm(2)). Additional dentures were treated only with PS (P+L-) or light (P-L+) or neither (P-L-). Samples of serial dilutions were spread on Sabouraud dextrose agar and incubated at 37°C for 48 h. The colonies were counted and the values of log (cfu/mL) were analyzed by Kruskall-Wallis and Dunn tests (p<0.05). RESULTS For all species of Candida, PDT resulted in significant reduction (p<0.05) of cfu/mL values from dentures when compared with P-L- (reductions from 1.73 to 3.99 log(10)). Significant differences (p<0.05), but lower reductions, were also observed for P+L- and P-L+when compared with P-L- for some species of Candida. CONCLUSIONS PDT was an effective method for reducing Candida spp. on dentures.

Prevalence of Candida spp. associated with bacteria species on complete dentures

OBJECTIVE This study evaluated the prevalence of Candida spp., Streptococcus mutans and Staphylococcus aureus isolated from complete upper dentures. BACKGROUND Factors that may increase the number of microorganisms in the oral cavity include denture-wearing and deficient denture hygiene. MATERIALS AND METHODS Ninety denture wearers participated in this study. Collection of biological material from each denture was performed on all denture surfaces. Conventional biochemical methods were used to identify the microorganisms. Descriptive statistical analysis was performed on percentage data. RESULTS Candida spp. were the most frequent microorganisms found on dentures (65.5%), while Strep. mutans and Staph. aureus were present in 53.3 and 34.4%, respectively. CONCLUSION Candida species were the most frequent microorganisms on dentures, followed by Strep. mutans and Staph. aureus.