Isolation of Candida spp. from denture-related stomatitis in Pará, Brazil

ABSTRACT The aim of this study was to isolate and identify Candida species from the oral cavity of denture wearers with denture-related stomatitis who were attended at the University Federal of Pará (Belém City, Pará State, Brazil). A total of 36 denture wearers with denture-related stomatitis were included, and type I (50%), type II (33%) and type III (17%) stomatitis were observed. Candida spp. were isolated from 89% of the cases and included five different Candida species. C. albicans was the most frequently recovered species (78% of the cases), followed by C. famata and C. tropicalis. We observed a significant association between Candida species isolation and unsatisfactory denture condition (p = 0.0017). Our results demonstrated the highly frequency of Candida species isolation in denture wearers with denture-related stomatitis and showed the relationship between these species and poor denture maintenance.

Alkaline Peroxides Versus Sodium Hypochlorite for Removing Denture Biofilm: a Crossover Randomized Trial

Abstract This study evaluated the efficacy of cleanser solutions on denture biofilm removal by a crossover randomized clinical trial. Thirty two edentulous patients were instructed to brush their dentures (specific brush and liquid soap) three times a day (after breakfast, lunch and dinner) and to soak them (≥ 8 h) in: (C) control -water; (AP): alkaline peroxide; or (SH) 0.5% sodium hypochlorite. Each solution was used for 21 days (three cycles of 7 days). At the end of each cycle, the inner surfaces of maxillary dentures were disclosed (1% neutral red) and photographed (HX1 - Sony). Areas (total and stained biofilm) were measured (Image Tool software) and the percentage of biofilm calculated as the ratio between the area of the biofilm multiplied by 100 and total surface area of the internal base of the denture. Data were compared by means of generalized estimating equation (α=5%) and multiple comparisons (Bonferroni; α=1.67%). Immersion in SH reduced biofilm (%) (8.3 ± 13.3B) compared to C (18.2 ± 14.9A) and AP (18.2 ± 16.6A). The 0.5% sodium hypochlorite solution was the most efficacious for biofilm removal. Alkaline peroxides may not lead to further biofilm removal in patients with adequate denture maintenance habits.

Resumo Este estudo avaliou a eficácia de soluções higienizadoras na remoção do biofilme de dentadura por meio de ensaio clínico randomizado cruzado. Trinta e dois pacientes desdentados foram instruídos a escovar suas dentaduras (escova específica e sabão líquido) três vezes ao dia (após café da manhã, almoço e jantar) e imergi-las (≥ 8 horas) em: (C) controle - água; (PA): peróxido alcalino; ou (HS) hipoclorito de sódio a 0,5%. Cada solução foi usada por 21 dias (três ciclos alternados de 7 dias). Ao final de cada ciclo, a superfície interna da dentadura maxilar foi evidenciada (vermelho neutro 1%) e fotografada (HX1- Sony). As áreas (total e corada com biofilme) foram medidas (software Image Tool), e a porcentagem de biofilme calculada como a relação entre a área do biofilme multiplicado por 100 e área da superfície total da base interna da dentadura. Os dados foram comparados por meio de equações de estimação generalizadas (α=5%) e comparações múltiplas (Bonferroni - α=1,67%). A imersão em HS reduziu o biofilme (%) (8,3 ± 13,3B) em comparação com C (18,2 ± 14,9A) e PA (18,2 ± 16,6A). A solução de hipoclorito de sódio a 0,5% foi a mais eficaz na remoção do biofilme. Peróxidos alcalinos podem não levar a maior remoção do biofilme em pacientes com hábitos adequados de manutenção de dentadura.

Antimicrobial action of sodium hypochlorite and castor oil solutions for denture cleaning – in vitro evaluation

The objective of this in vitro study was to evaluate the antimicrobial action of sodium hypochlorite (0.25% and 0.50%) and 10% castor oil solutions against specific microorganisms, by counting Colony Forming Units (CFU) of clinically important bacteria and Candida species. Acrylic resin specimens (n = 320; Lucitone 550) were obtained from square metal matrices (10 x 10 x 2 mm), sterilized by microwave (650W, for 6 minutes) and contaminated by Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, Bacillus subtilis, Escherichia coli, Streptococcus mutans, Enterococcus faecalisand Candida glabrata. The specimens were immersed for 20 minutes in one of the following hygiene solutions (n = 10/each): A – 0.25% Sodium hypochlorite; B – 0.5% Sodium hypochlorite; C – 10% Castor oil solution; and D (Control) – saline. Adhered cells were suspended and inoculated into a selective solid medium (37ºC for 24 h). The Student’s t-test (α = 0.05) was performed to compare log10(CFU+1)/mL between Groups C and D. The results showed that sodium hypochlorite (0.25% and 0.5%) completely eliminated all detectable microorganisms. The castor oil solution eliminatedB. subtilisand reduced counts for other strains. Differences between C and D were significant (p < 0.05) for all species except for E. faecalis. Both sodium hypochlorite solutions (0.25% and 0.5%) were effective in eliminating all microorganisms evaluated, and may be useful as cleaning solutions for complete dentures. The castor oil solution provided moderate efficacy and performed differently on the tested species, with the strongest effect on B. subtilis and with non-significant action on E. faecalis.

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

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

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

Comparative study of Candida by conventional and CHROMagar method in non-denture and denture wearers by oral rinse technique.

Introduction: Candidal species colonizes the oral cavities of healthy individuals without dentures and also of denture wearers. Soft liners and tissue conditioning materials have been found to support the growth of Candida albicans which may predispose to lesions. The most important and common candidal species are C. albicans, C. tropicalis, and C. glabrata. C albicans is usually isolated from both the fitting surface of the denture and the denture-bearing mucosa of the affected patients. The aim of this study was to isolate, quantify, and speciate candidal species in non-denture wearers (controls) and denture wearers (study group) by the oral rinse technique. Isolation was done using Sabouraud dextrose agar (SDA). Speciation was done using conventional methods like the germ tube test, carbohydrate fermentation test, urease test, as well as the CHROMagar method. Aims and Objective: 1) To assess the prevalence of Candida in non-denture wearers and in denture wearers by oral rinse technique, with isolation on SDA; 2) to speciate and quantify Candida in non-denture wearers and denture wearers by using conventional methods (germ tube test, carbohydrate fermentation test, urease test) and the CHROMagar method; 3) to assess the influence of smoking and diabetes on candidal species among the denture wearers; and 4) to assess the sensitivity and specificity of SDA and CHRO Magar Materials and Methods: Salivary samples for Candida evaluation were collected from the subjects in sterile sample containers, using the oral rinse technique. Results: C glabrata was the most commonly found species among denture wearers and non-denture wearers both by conventional and CHROMagar methods. In males, C. albicans was the predominant species, whereas C. glabrata was the predominant species in females. Candidal colonization was higher in denture wearers compared to non-denture wearers, especially among females. The CHROMagar method was more rapid compared to conventional methods. In the present study, CHROMagar Candida showed 100% specificity and 100% sensitivity when compared to SDA and conventional methods.

Multi-species biofilm of Candida albicans and non-Candida albicans Candida species on acrylic substrate

OBJECTIVE: In polymicrobial biofilms bacteria extensively interact with Candida species, but the interaction among the different species of the Candida is yet to be completely evaluated. In the present study, the difference in biofilm formation ability of clinical isolates of four species of Candida in both single-species and multi-species combinations on the surface of dental acrylic resin strips was evaluated. MATERIAL AND METHODS: The species of Candida, isolated from multiple species oral candidiasis of the neutropenic patients, were used for the experiment. Organisms were cultured on Sabouraud dextrose broth with 8 percent glucose (SDB). Biofilm production on the acrylic resins strips was determined by crystal violet assay. Student's t-test and ANOVA were used to compare in vitro biofilm formation for the individual species of Candida and its different multi-species combinations. RESULTS: In the present study, differences between the mean values of the biofilm-forming ability of individual species (C. glabrata>C. krusei>C. tropicalis>C. albicans) and in its multi-species' combinations (the highest for C. albicans with C. glabrata and the lowest for all the four species combination) were reported. CONCLUSIONS: The findings of this study showed that biofilm-forming ability was found greater for non-Candida albicans Candida species (NCAC) than for C. albicans species with intra-species variation. Presence of C. albicans in multi-species biofilms increased, whereas; C. tropicalis decreased the biofilm production with all other NCAC species.

Biochemically assessed pathological activity of oral candida in denture and non denture wearers

To assess the Candida activity through recording pH changes in Candida selective medium inoculated with standardized dilution salivary samples obtained from dentures wearers and non denture wearers. A Cross Sectional, Comparative study during March-September 2007 was conducted at Khyber College of Dentistry Peshawar and Zoology Department, University of Peshawar [Pakistan]. Using a structured proforma, data were collected from 100 subjects [50 DWs and 50 NDWs]. Stimulated salivary samples were collected from each subject. One ml of standard dilution saliva was mixed with 4 ml sterile liquid Candida-selective medium. The extent of Candida activity was then determined, biochemically, through change in pH of the medium 24 hrs after inoculation and incubation at 37°C. The influence of age, gender, socioeconomic status, smoking habits, medication-use, oral-hygiene maintenance method, denture-plaque level, and level of salivary function was also investigated. Pathological Candida activity was seen both in DWs and NDWs. However, it was significantly higher in DWs [p value = 0.00] with mean pH decrease of 1.9 as compared to 0.8 in NDWs. Except age, the effect of gender, socioeconomic status, medications, oral hygiene maintenance on the oral Candida activity was insignificant both within and between groups. Not-with-standing the varying effect of other variables, denture-wearing significantly enhanced pathogenic activity of oral Candida. The importance of meticulous oral and denture hygiene maintenance and rest to denture-supporting tissues is re-emphasized