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 (107 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.

Daily biofilm control and oral health: Consensus on the epidemiological challenge-Latin American Advisory Panel

Our understanding of dental plaque biofilm has evolved since the nonspecific plaque hypothesis that considered plaque as a nonspecific mass of native microorganisms that, because of lack of oral hygiene, builds up in proportions great enough to overcome the host resistance threshold and affect the tooth structure and tooth supporting tissues. A great diversity of microorganisms-over 700 species-was detected in the oral cavity, and evidence shows that the investigation of specific microorganisms or associations of microorganisms as etiological agents for periodontal diseases and caries is not a simplistic approach. Although clinical evidence shows that oral mechanical hygiene is fundamental to prevent and control caries and periodontal disease, it is important to highlight that optimal control is not achieved by most individuals. Thus the complementary use of chemotherapeutic agents has been investigated as a way to overcome the deficiencies of mechanical oral hygiene habits, insofar as they reduce both plaque formation and gingival inflammation, and represent a valid strategy to change the biofilm and maintain dental and periodontal health. The role of the dental professional is to monitor patients and offer them the best recommendations to preserve oral health throughout their life. With this in mind, chemical control should be indicated as part of daily oral hygiene, together with mechanical procedures, for all individuals who present supragingival and/or subgingival biofilm, taking into account age, physical and/or psychological limitations, allergies, and other factors.

Effect of experimental photopolymerized coatings on the hydrophobicity of a denture base acrylic resin and on Candida albicans adhesion

Objective: This study investigated the effect of experimental photopolymerized coatings, containing zwitterionic or hydrophilic monomers, on the hydrophobicity of a denture base acrylic resin and on Candida albicans adhesion. Methods: Acrylic specimens were prepared with rough and smooth surfaces and were either left untreated (control) or coated with one of the following experimental coatings: 2-hydroxyethyl methacrylate (HE); 3-hydroxypropyl methacrylate (HP); and 2-trimethylammonium ethyl methacrylate chloride (T); and sulfobetaine methacrylate (S). The concentrations of these constituent monomers were 25%, 30% or 35%. Half of the specimens in each group (control and experimentals) were coated with saliva and the other half remained uncoated. The surface free energy of all specimens was measured, regardless of the experimental condition. C. albicans adhesion was evaluated for all specimens, both saliva conditioned and unconditioned. The adhesion test was performed by incubating specimens in C. albicans suspensions (1 × 10 7 cell/mL) at 37 °C for 90 min. The number of adhered yeasts were evaluated by XTT (2,3-bis[2-methoxy-4-nitro-5-sulfophenyl]-5-[{phenylamino} carbonyl]-2H-tetrazolium-hydroxide) method. Results: For rough surfaces, coatings S (30 or 35%) and HP (30%) resulted in lower absorbance values compared to control. These coatings exhibited more hydrophilic surfaces than the control group. Roughness increased the adhesion only in the control group, and saliva did not influence the adhesion. The photoelectron spectroscopy analysis (XPS) confirmed the chemical changes of the experimental specimens, particularly for HP and S coatings. Conclusions: S and HP coatings reduced significantly the adhesion of C. albicans to the acrylic resin and could be considered as a potential preventive treatment for denture stomatitis. © 2012 Elsevier Ltd.

In vitro antimicrobial activity of auxiliary chemical substances and natural extracts on Candida albicans and Enterococcus faecalis in root canals

Objective: The aim of this study was to evaluate the antimicrobial activity of auxiliary chemical substances and natural extracts on Candida albicans and Enterococcus faecali's inoculated in root canals. Material and Methods: Seventy-two human tooth roots were contaminated with C. albicans and E. faecalis for 21 days. The groups were divided according to the auxiliary chemical substance into: G1) 2.5% sodium hypochlorite (NaOCl), G2) 2% chlorhexidine gel (CHX), G3) castor oil, G4) glycolic Aloe vera extract, g5) glycolic ginger extract, and G6) sterile saline (control). The samples of the root canal were collected at different intervals: confirmation collection, at 21 days after contamination; 1st collection, after instrumentation; and 2nd collection, seven days after instrumentation. Microbiological samples were grown in culture medium and incubated at 37°C for 48 hours. Results: The results were submitted to the Kruskal-Wallis and Dunn (5%) statistical tests. NaOCl and CHX completely eliminated the microorganisms of the root canals. Castor oil and ginger significantly reduced the number of CFU of the tested bacteria. Reduction of CFU/mL at the 1st and 2nd collections for groups G1, G2, G3 and G4 was greater in comparison to groups G5 and G6. Conclusion: It was concluded that 2.5% sodium hypochlorite and 2% chlorhexidine gel were more effective in eliminating C. albicans and E. faecalis, followed by the castor oil and glycolic ginger extract. The Aloe vera extract showed no antimicrobial activity.

Antimicrobial activity of the synthetic peptide Lys-a1 against oral streptococci

The peptide LYS-[TRP6]-Hy-A1 (Lys-a1) is a synthetic derivative of the peptide Hy-A1, initially isolated from the frog species Hypsiboas albopunctatus. According to previous research, it is a molecule with broad antimicrobial activity. The objective of this study was to evaluate the antimicrobial activity of the synthetic peptide Lys-a1 (KIFGAIWPLALGALKNLIK- NH2) on the planktonic and biofilm growth of oral bacteria. The methods used to evaluate antimicrobial activity include the following: determination of the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) in microtiter plates for growth in suspension and quantification of biomass by crystal violet staining and counting of colony forming units for biofilm growth. The microorganisms Streptococcus oralis, Streptococcus sanguinis, Streptococcus parasanguinis, Streptococcus salivarius, Streptococcus mutans and Streptococcus sobrinus were grown in Brain Heart Infusion broth at 37 °C under atmospheric pressure with 10% CO2. The peptide was solubilized in 0.1% acetic acid (v/v) at various concentrations (500-1.9 μg mL-1). Chlorhexidine gluconate 0.12% was used as the positive control, and BHI culture medium was used as the negative control. The tested peptide demonstrated a remarkable antimicrobial effect, inhibiting the planktonic and biofilm growth of all strains tested, even at low concentrations. Thus, the peptide Lys-a1 is an important source for potential antimicrobial agents, especially for the control and prevention of microbial biofilms, which is one of the most important factors in cariogenic processes. © 2012 Elsevier Inc.

Antifungal activity of silver nanoparticles in combination with nystatin and chlorhexidine digluconate against Candida albicans and Candida glabrata biofilms

Although silver nanoparticles (SN) have been investigated as an alternative to conventional antifungal drugs in the control of Candida-associated denture stomatitis, the antifungal activity of SN in combination with antifungal drugs against Candida biofilms remains unknown. Therefore, the aim of this study was to evaluate the antifungal efficacy of SN in combination with nystatin (NYT) or chlorhexidine digluconate (CHG) against Candida albicans and Candida glabrata biofilms. The drugs alone or combined with SN were applied on mature Candida biofilms (48 h), and after 24 h of treatment their antibiofilm activities were assessed by total biomass quantification (by crystal violet staining) and colony forming units enumeration. The structure of Candida biofilms was analysed by scanning electron microscopy (SEM) images. The data indicated that SN combined with either NYT or CHG demonstrated synergistic antibiofilm activity, and this activity was dependent on the species and on the drug concentrations used. SEM images showed that some drug combinations were able to disrupt Candida biofilms. The results of this study suggest that the combination of SN with NYT or CHG may have clinical implications in the treatment of denture stomatitis. However, further studies are needed before recommending the use of these drugs safely in clinical situations. © 2013 Blackwell Verlag GmbH.

Recent mouse and rat methods for the study of experimental oral candidiasis

The Candida genus expresses virulence factors that, when combined with immunosuppression and other risk factors, can cause different manifestations of oral candidiasis. The treatment of mucosal infections caused by Candida and the elucidation of the disease process have proven challenging. Therefore, the study of experimentally induced oral candidiasis in rats and mice is useful to clarify the etiopathology of this condition, improve diagnosis, and search for new therapeutic options because the disease process in these animals is similar to that of human candidiasis lesions. Here, we describe and discuss new studies involving rat and mouse models of oral candidiasis with respect to methods for inducing experimental infection, methods for evaluating the development of experimental candidiasis, and new treatment strategies for oral candidiasis. © 2013 Landes Bioscience.

Effect of diamond-like carbon thin film coated acrylic resin on candida albicans biofilm formation

Purpose: The purpose of this study was to evaluate the effect of diamond-like carbon thin films doped and undoped with silver nanoparticles coating poly(methyl methacrylate) (PMMA) on Candida albicans biofilm formation. The control of biofilm formation is important to prevent oral diseases in denture users. Materials and Methods: Forty-five PMMA disks were obtained, finished, cleaned in an ultrasonic bath, and divided into three groups: Gc, no surface coating (control group); Gdlc, coated with diamond-like carbon film; and Gag, coated with diamond-like carbon film doped with silver nanoparticles. The films were deposited using a reactive magnetron sputtering system (physical vapor deposition process). The specimens were characterized by optical profilometry, atomic force microscopy, and Rutherford backscattering spectroscopy analyses that determined differences in chemical composition and morphological structure. Following sterilization of the specimens by γ-ray irradiation, C. albicans (ATCC 18804) biofilms were formed by immersion in 2 ml of Sabouraud dextrose broth inoculated with a standardized fungal suspension. After 24 hours, the number of colony forming units (cfu) per specimen was counted. Data concerning biofilm formation were analyzed using ANOVA and the Tukey test (p < 0.05). Results: C. albicans biofilm formation was significantly influenced by the films (p < 0.00001), reducing the number of cfu, while not affecting the roughness parameters (p > 0.05). The Tukey test showed no significant difference between Gdlc and Gag. Films deposited were extremely thin (∼50 nm). The silver particles presented a diameter between 60 and 120 nm and regular distribution throughout the film surface (to Gag). Conclusion: Diamond-like carbon films, doped or undoped with silver nanoparticles, coating the base of PMMA-based dentures could be an alternative procedure for preventing candidosis in denture users. © 2013 by the American College of Prosthodontists.

Cytotoxicity of Brazilian plant extracts against oral microorganisms of interest to dentistry

Background: With the emergence of strains resistant to conventional antibiotics, it is important to carry studies using alternative methods to control these microorganisms causing important infections, such as the use of products of plant origin that has demonstrated effective antimicrobial activity besides biocompatibility. Therefore, this study aimed to evaluate the antimicrobial activity of plant extracts of Equisetum arvense L., Glycyrrhiza glabra L., Punica granatum L. and Stryphnodendron barbatimam Mart. against Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus mutans, Candida albicans, Candida tropicalis, and Candida glabrata, and to analyze the cytotoxicity of these extracts in cultured murine macrophages (RAW 264.7).Methods: Antimicrobial activity of plant extracts was evaluated by microdilution method based on Clinical and Laboratory Standards Institute (CLSI), M7-A6 and M27-A2 standards. The cytotoxicity of concentrations that eliminated the microorganisms was evaluated by MTT colorimetric method and by quantification of proinflammatory cytokines (IL-1β and TNF-α) using ELISA.Results: In determining the minimum microbicidal concentration, E. arvense L., P. granatum L., and S. barbatimam Mart. extracts at a concentration of 50 mg/mL and G. glabra L. extract at a concentration of 100 mg/mL, were effective against all microorganisms tested. Regarding cell viability, values were 48% for E. arvense L., 76% for P. granatum L, 86% for S. barbatimam Mart. and 79% for G. glabra L. at the same concentrations. About cytokine production after stimulation with the most effective concentrations of the extracts, there was a significant increase of IL-1β in macrophage cultures treated with S. barbatimam Mart. (3.98 pg/mL) and P. granatum L. (7.72 pg/mL) compared to control (2.20 pg/mL) and a significant decrease of TNF-α was observed in cultures treated with G. glabra L. (4.92 pg/mL), S. barbatimam Mart. (0.85 pg/mL), E. arvense L. (0.83 pg/mL), and P. granatum L. (0.00 pg/mL) when compared to control (41.96 pg/mL).Conclusions: All plant extracts were effective against the microorganisms tested. The G. glabra L. extract exhibited least cytotoxicity and the E. arvense L. extract was the most cytotoxic. © 2013 de Oliveira et al.; licensee BioMed Central Ltd.

Comparison of the effect of rose bengal- and eosin Y-mediated photodynamic inactivation on planktonic cells and biofilms of Candida albicans

Candida albicans is an opportunistic yeast that can cause oral candidosis through the formation of a biofilm, an important virulence factor that compromises the action of antifungal agents. The objective of this study was to compare the effect of rose bengal (RB)- and eosin Y (EY)-mediated photodynamic inactivation (PDI) using a green light-emitting diode (LED; 532 ± 10 nm) on planktonic cells and biofilms of C. albicans (ATCC 18804). Planktonic cultures were treated with photosensitizers at concentrations ranging from 0.78 to 400 μM, and biofilms were treated with 200 μM of photosensitizers. The number of colony-forming unit per milliliter (CFU/mL) was compared by analysis of variance and Tukey's test (P ≤ 0.05). After treatment, one biofilm specimen of the control and PDI groups were examined by scanning electron microscopy. The photosensitizers (6.25, 25, 50, 200, and 400 μM of EY, and 6.25 μM of RB or higher) significantly reduced the number of CFU/mL in the PDI groups when compared to the control group. With respect to biofilm formation, RB- and EY-mediated PDI promoted reductions of 0.22 log10 and 0.45 log10, respectively. Scanning electron microscopy showed that the two photosensitizers reduced fungal structures. In conclusion, EY- and RB-mediated PDI using LED irradiation significantly reduced C. albicans planktonic cells and biofilms. © 2013 Springer-Verlag London.

Photodynamic inactivation of clinical isolates of Candida using Photodithazine

This study evaluated the photodynamic inactivation (PDI) mediated by Photodithazine® (PDZ) against 15 clinical isolates of Candida albicans, Candida glabrata and Candida tropicalis. Each isolate, in planktonic and biofilm form, was exposed to PDI by assessing a range of PDZ concentrations and light emitting diode fluences. Cell survival of the planktonic suspensions was determined by colony forming units (CFU ml-1). The antifungal effects of PDI against biofilms were evaluated by CFU ml-1 and metabolic assay. Data were analyzed by non-parametric tests (α = 0.05). Regardless of the species, PDI promoted a significant viability reduction of planktonic yeasts. The highest reduction in cell viability of the biofilms was equivalent to 0.9 log10 (CFU ml-1) for C. albicans, while 1.4 and 1.5 log10 reductions were obtained for C. tropicalis and C. glabrata, respectively. PDI reduced the metabolic activity of biofilms by 62.1, 76.0, and 76.9% for C. albicans, C. tropicalis, and C. glabrata, respectively. PDZ-mediated PDI promoted significant reduction in the viability of Candida isolates. © 2013 Taylor & Francis.

Efeito fotodinâmico da curcumina em micelas de cetrimida sobre cepas de candida susceptíveis e resistentes a fluconazol

Pós-graduação em Biociências e Biotecnologia Aplicadas à Farmácia - FCFAR

Efeitos da terapia fotodinâmica sobre cepas de Candida isoladas de pacientes submetidos à antibioticoterapia prolongada

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Prevalência e susceptibilidade aos antifúngicos de isolados de leveduras do gênero candida da cavidade bucal de pacientes com hanseníase

Pós-graduação em Biopatologia Bucal - ICT

Cordia verbenacea DC: perfil morfo-anatômico, histoquímico, farmacognóstico e avaliação da atividade anti-candida do extrato hidroetanólico e suas frações

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)