Effectiveness of experimental dentifrices based on essential oils on biofilm on complete dentures: an in vitro study.

Specific products containing natural resources can contribute to the innovation of complete denture hygiene. OBJECTIVE: To conduct an in vitro evaluation of experimental dentifrices containing essential oils of Bowdichia virgilioides Kunth (BvK), Copaifera officinalis (Co), Eucalyptus citriodora (Ec), Melaleuca alternifolia (Ma) and Pinus strobus (Ps) at 1%. METHODOLOGY: The variables evaluated were organoleptic and physicochemical characteristics, abrasiveness (mechanical brushing machine) simulating 2.5 years, and microbial load (Colony Forming Units - CFU/mL), metabolic activity (XTT assay) and cell viability (Live/Dead® BacLight™ kit) of the multispecies biofilm (Streptococcus mutans: Sm, Staphylococcus aureus: Sa, Candida albicans: Ca and Candida glabrata: Cg). Specimens of heat-polymerized acrylic resins (n=256) (n=96 specimens for abrasiveness, n=72 for microbial load count, n=72 for biofilm metabolic activity, n=16 for cell viability and total biofilm quantification) with formed biofilm were divided into eight groups for manual brushing (20 seconds) with a dental brush and distilled water (NC: negative control), Trihydral (PC: positive control), placebo (Pl), BvK, Co, Ec, Ma or Ps. After brushing, the specimens were washed with PBS and immersed in Letheen Broth medium, and the suspension was sown in solid specific medium. The organoleptic characteristics were presented by descriptive analysis. The values of density, pH, consistency and viscosity were presented in a table. The data were analyzed with the Wald test in a generalized linear model, followed by the Kruskal-Wallis test, Dunn's test (mass change) and the Bonferroni test (UFC and XTT). The Wald test in Generalized Estimating Equations and the Bonferroni test were used to analyze cell viability. RESULTS: All dentifrices showed stable organoleptic characteristics and adequate physicochemical properties. CN, Ec, Ps, Pl and PC showed low abrasiveness. There was a significant difference between the groups (p<0.001) for microbial load, metabolic activity and biofilm viability. CONCLUSIONS: It was concluded that the BvK, Ec and Ps dentifrices are useful for cleaning complete dentures, as they have antimicrobial activity against biofilm. The dentifrices containing Bowdichia virgilioides Kunth showed medium abrasiveness and should be used with caution.

Evaluation of surface roughness, wettability and adhesion of multispecies biofilm on 3D-printed resins for the base and teeth of complete dentures.

OBJECTIVE: This study evaluated the surface roughness, wettability and adhesion of multispecies biofilms (Candida albicans, Staphylococcus aureus and Streptococcus mutans) on 3D-printed resins for complete denture bases and teeth compared to conventional resins (heat-polymerized acrylic resin; artificial pre-fabricated teeth). METHODOLOGY: Circular specimens (n=39; 6.0 mm Ø × 2.0 mm) of each group were subjected to roughness (n=30), wettability (n=30) and biofilm adhesion (n=9) tests. Three roughness measurements were taken by laser confocal microscopy and a mean value was calculated. Wettability was evaluated by the contact angle of sessile drop method, considering the mean of the three evaluations per specimen. In parallel, microorganism adhesion to resin surfaces was evaluated using a multispecies biofilm model. Microbial load was evaluated by determining the number of Colony Forming Units (CFU/mL) and by scanning electron microscopy (SEM). Data were subjected to the Wald test in a generalized linear model with multiple comparisons and Bonferroni adjustment, as well as two-way ANOVA (α=5%). RESULTS: The roughness of the conventional base resin (0.01±0.04) was lower than that of the conventional tooth (0.14±0.04) (p=0.023) and 3D-printed base (0.18±0.08) (p<0.001). For wettability, conventional resin (84.20±5.57) showed a higher contact angle than the 3D-printed resin (60.58±6.18) (p<0.001). Higher microbial loads of S. mutans (p=0.023) and S. aureus (p=0.010) were observed on the surface of the conventional resin (S. mutans: 5.48±1.55; S. aureus: 7.01±0.57) compared to the 3D-printed resin (S. mutans: 4.11±1.96; S. aureus: 6.42±0.78). The adhesion of C. albicans was not affected by surface characteristics. The conventional base resin showed less roughness than the conventional dental resin and the printed base resin. CONCLUSION: The 3D-printed resins for base and tooth showed less hydrophobicity and less adhesion of S. mutans and S. aureus than conventional resins.

Relation between the risk factors for the severity of denture stomatitis and quality of life of complete edentulous individuals: a cross-sectional study.

OBJECTIVE: To assess the association between risk factors for developing denture stomatitis (DS) and oral health-related quality of life (OHRQoL) in complete denture wearers. METHODOLOGY: Participants of both sexes, wearing complete dentures, were classified using the modified Newton classification for the absence or the severity of DS and allocated to groups Normal or zero, IA, IB, II, and III. Lifestyle, oral and denture history, and medication use were assessed using specific questionnaires; clinical parameters such as anatomical characteristics of support were evaluated with the Kapur classification; salivary flow (SF) was calculated by the volume of unstimulated saliva per minute; and microbial load was determined by counting colony forming units (CFU) of target microorganisms present in the biofilm collected from dentures and palate. OHIP-EDENT assessed the OHRQoL. Kendall's tau_b and Spearman tests were applied with a significance level of 5%. RESULTS: 184 patients (143 female and 41 male) aged 65.5 ± 6.8 years were evaluated. Positive correlations were found for sex (women; p=0.013, r=0.16), individuals who started to consume alcoholic beverages as a young adult (18-27 years) (p=0.008, r=0.22), CFU of Candida spp. (p<0.001, r=0.27 denture; p<0.001, r=0.31 palate); Candida albicans (p=0.004, r=0.22 denture; p=0.003, r=0.25 palate), and Candida glabrata (p=0.004, r=0.22 denture; p=0.001, r=0.27 palate). Moreover, negative correlations with DS were found for CFU of Staphylococcus spp. (p=0.004, r=-0.20 palate) and enterobacteria (p=0.002, r=-0.24 palate), as well as a negative correlation between SF (p=0.009, r=-0.193) and DS. The CFU of Staphylococcus spp. and enterobacteria on the palate significantly correlated with OHRQoL. CONCLUSION: Being female, consuming alcoholic beverages as a young adult, CFU of Candida spp., Candida albicans, Candida glabrata, and salivary flow may be the most significant risk factors for DS. The microbial load of Staphylococcus spp. and enterobacteria seems to influence the quality of life for complete denture wearers.

Effect of Hygiene Protocols on the Mechanical and Physical Properties of Two 3D-Printed Denture Resins Characterized by Extrinsic Pigmentation as Well as the Mixed Biofilm Formed on the Surface.

To assess the effect of hygiene protocols and time on the physical-mechanical properties and colony-forming units (CFU) of Candida albicans, Staphylococcus aureus, and Streptococcus mutans on 3D-printed denture resins (SmartPrint and Yller) with extrinsic pigmentation compared to conventional resin (CR). The protocols were evaluated: brushing (B), brushing and immersion in water (W), 0.25% sodium hypochlorite (SH), and 0.15% triclosan (T), simulating 0, 1, 3, and 5 years. The data were analyzed by ANOVA with repeated measurements, ANOVA (Three-way) and Tukey's post-test, generalized linear model with Bonferroni adjustment, and ANOVA (Two-way) and Tukey's post-test (α = 0.05). The protocols influenced color (p = 0.036) and Knoop hardness (p < 0.001). Surface roughness was influenced by protocols/resin (p < 0.001) and time/resin (p = 0.001), and flexural strength by time/protocols (p = 0.014). C. albicans showed interactions with all factors (p = 0.033). Staphylococcus aureus was affected by protocols (p < 0.001). Streptococcus mutans exhibited no count for SH and T (p < 0.001). Yller resin showed more color changes. The 3D-printed resins displayed lower microhardness, increased roughness, and decreased flexural strength compared to CR with all protocols in a simulated period of 5 years. The indication of printed resins should be restricted to less than 3 years.

Relation between the risk factors for the severity of denture stomatitis and quality of life of complete edentulous individuals: a cross-sectional study

Abstract Objective To assess the association between risk factors for developing denture stomatitis (DS) and oral health-related quality of life (OHRQoL) in complete denture wearers. Methodology Participants of both sexes, wearing complete dentures, were classified using the modified Newton classification for the absence or the severity of DS and allocated to groups Normal or zero, IA, IB, II, and III. Lifestyle, oral and denture history, and medication use were assessed using specific questionnaires; clinical parameters such as anatomical characteristics of support were evaluated with the Kapur classification; salivary flow (SF) was calculated by the volume of unstimulated saliva per minute; and microbial load was determined by counting colony forming units (CFU) of target microorganisms present in the biofilm collected from dentures and palate. OHIP-EDENT assessed the OHRQoL. Kendall's tau_b and Spearman tests were applied with a significance level of 5%. Results 184 patients (143 female and 41 male) aged 65.5 ± 6.8 years were evaluated. Positive correlations were found for sex (women; p=0.013, r=0.16), individuals who started to consume alcoholic beverages as a young adult (18-27 years) (p=0.008, r=0.22), CFU of Candida spp. (p<0.001, r=0.27 denture; p<0.001, r=0.31 palate); Candida albicans (p=0.004, r=0.22 denture; p=0.003, r=0.25 palate), and Candida glabrata (p=0.004, r=0.22 denture; p=0.001, r=0.27 palate). Moreover, negative correlations with DS were found for CFU of Staphylococcus spp. (p=0.004, r=-0.20 palate) and enterobacteria (p=0.002, r=-0.24 palate), as well as a negative correlation between SF (p=0.009, r=-0.193) and DS. The CFU of Staphylococcus spp. and enterobacteria on the palate significantly correlated with OHRQoL. Conclusion Being female, consuming alcoholic beverages as a young adult, CFU of Candida spp., Candida albicans, Candida glabrata, and salivary flow may be the most significant risk factors for DS. The microbial load of Staphylococcus spp. and enterobacteria seems to influence the quality of life for complete denture wearers.

Effect of a Hygiene Protocol on Denture-Related Stomatitis Remission, Local Inflammatory Factors, and Hemodynamic Responses by Arterial Pressure.

Denture-related stomatitis (DRS) is frequent oral inflammation in complete denture wearers. This study evaluated the effect of a hygiene protocol on DRS remission, local inflammatory factors, and hemodynamic responses. Thirty-three individuals were enrolled in the study. The outcomes were measured before and after 10 days of a hygiene protocol treatment consisting of brushing the palate with a soft brush and water and denture brushing with a denture-specific brush and mild soap, as well as immersion of the denture for 20 min in a 0.25% sodium hypochlorite solution. Data were analyzed by paired Wilcoxon for biofilm removal and CFU count of microorganisms. The paired T test was used to assess salivary MUC 1, cytokines, and arterial pressure (p < 0.05). A significant difference was found in the DRS degree (p < 0.001), biofilm (p < 0.001), microbial load of Candida spp. (p < 0.001), Gram-negative (p < 0.004), Staphylococcus spp. (p < 0.001), and S. mutans (p < 0.001) of the denture, and S. mutans (p < 0.001) of the palate after use of the protocol. The salivary flow (p = 0.2) and pH (p = 0.97) did not change; there was an increase of MUC 1 (p = 0.049) and a decrease in IL-6 (p = 0.038), IL-2 (p = 0.04), IL-10 (p = 0.041), and IFNγ (p = 0.04). There was also a decrease in systolic (p = 0.012) and mean arterial pressure (p = 0.02). The current hygiene protocol reduced the inflammation degree of DRS and promoted an improvement of local inflammatory factors and a reduction in the systolic arterial pressure of the patients.

Influence of glucose supplementation on biofilm formation of Candida albicans and Candida glabrata isolated from diabetic and non-diabetic individuals.

OBJECTIVE: This study evaluated the effect of different glucose concentration on biofilm formation of Candida albicans and Candida glabrata strains isolated from diabetic and non-diabetic individuals. METHODS: The study was divided into two stages: (I) selection and identification of 48 C. albicans and C. glabrata strains by polymerase chain reaction followed by restriction fragment length polymorphism analysis (PCR/RFLP); (II) evaluation of biofilm formation by means of viability rates (colony-forming units), biofilm dry matter (mg) and biofilm-covered areas (µm2). Statistical comparisons were performed through nonparametric analysis of longitudinal data in factorial experiments with pairwise comparisons using Friedman Conover's test (α = 0.05). RESULTS: All the Candida spp. had their identifications confirmed by PCR/RFLP. C. albicans biofilm of strains from diabetic individuals cultivated in different glucose concentration showed higher viability rates than strains from non-diabetic individuals. No difference was observed on viability of C. glabrata biofilm. Regarding biofilm dry matter, C. albicans biofilm of strains from diabetic individuals cultivated in different glucose concentration showed lower amount in weight than strains from non-diabetic individuals. In C. glabrata strains, this result was only observed in biofilms cultivated with no glucose supplementation. With regard to biofilm-covered areas, only glucose supplementation and non-diabetic condition showed a positive effect on C. albicans biofilm development, and no condition affected C. glabrata biofilm formation. CONCLUSION: The strain type (C. albicans and C. glabrata) isolated from diabetic and non-diabetic individuals influenced on biofilm formation, but glucose supplementation did not.

Increased diversity, fungal burden, and virulence of oral Candida spp. in patients undergoing anti-tuberculosis treatment.

Some studies have demonstrated a high prevalence of Candida species in patients with tuberculosis (TB). This is most likely due to long-term antimicrobial therapy. To date, no longitudinal studies addressed the effects of anti-TB treatment on the fungal burden and virulence of Candida spp. This study investigated the prevalence and virulence of Candida spp. in the oral cavity of 30 TB patients at different stages of treatment through a cohort study. These results were compared with those of 60 systemically healthy individuals in a cross-sectional study. Oral rinse samples from TB patients were collected before 45 and after 120 days of treatment. In the control group, the biological samples were collected only once. Candida spp. were identified by restriction fragment length polymorphism (RFLP) assays, and the following virulence factors were studied: phospholipase C and proteinase production, as well as Candida spp. biofilm and hyphae formation. The clinical diagnosis of TB and its treatment time were associated with the greater fungal burden (p < 0.0001), presence of non-albicans Candida (NAC) species (p = 0.0003), and increased virulence factors when compared with the Candida spp. isolated from systemically healthy individuals. The results showed that anti-TB treatment time was responsible for the increased fungal burden and isolation of NAC in TB patients (p = 0.0233). The increased prevalence, quantification, and virulence of Candida spp. isolated from the oral cavity of TB patients highlight the greater risk of oral lesions and cases of systemic dissemination in these patients.

Complete denture hygiene solutions: antibiofilm activity and effects on physical and mechanical properties of acrylic resin.

BACKGROUND: Appropriated denture hygiene is a predictive factor for longevity of rehabilitation treatment and maintenance of the oral mucosal health. Although, disinfectant solutions are commonly used as denture cleansers, the impact of these solutions on acrylic resin-based dentures remain unclear. OBJECTIVE: To evaluate, in vitro, the antibiofilm activity of complete denture hygiene solutions and their effects on physical and mechanical properties of acrylic resin. METHODOLOGY: For antibiofilm activity measurement acrylic resin specimens were contaminated with Candida albicans, Candida glabrata and Streptococcus mutans. After biofilm growth, the specimens were assigned to the hygiene solutions: Distilled water (Control); 0.2% Sodium hypochlorite (SH); Efferdent Power Clean Crystals (EPC) and 6.25% Ricinus communis (RC). The viability of microorganisms was evaluated by agar plate counts. In parallel, physical, and mechanical properties of the acrylic resin were evaluated after simulating a 5-year period of daily immersion in the previously mentioned solutions. The changes in surface roughness, color, microhardness, flexural strength, impact strength, sorption and solubility were evaluated. Data were compared by ANOVA followed by the Tukey test or Kruskal-Wallis followed by the Dunn test depending on the distribution (α=0.05). RESULTS: Regarding antibiofilm action, SH eliminated all microorganisms while EPC and RC exhibited moderate action against S. mutans (p=0.001) and C. glabrata (p<0.001), respectively. Relative to effects on the physical and mechanical properties of the acrylic resin, RC led to higher values of color change (p=0.030), hardness (p<0.001), surface roughness (p=0.006) and flexural strength (p<0.001). Moreover, RC induced the highest values of changes in solubility (p<0.001). EPC promoted greater changes in surface morphology, whereas immersion in SH retained the initial appearance of the acrylic resin surface. All hygiene solutions reduced the impact strength (p<0.05). CONCLUSION: SH presented the most effective antibiofilm activity. In addition, changes on properties were observed after immersion in RC, which were considered within acceptable limits.

Action of disinfectant solutions on adaptive capacity and virulence factors of the Candida spp. biofilms formed on acrylic resin.

BACKGROUND: Understanding the behavior of Candida spp. when exposed to denture disinfectants is essential to optimize their effectiveness. Changes in the virulence factors may cause increased resistance of Candida spp. to disinfectant agents. OBJECTIVE: To evaluate the microbial load, cellular metabolism, hydrolytic enzyme production, hyphae formation, live cell and biofilm quantification of Candida albicans, Candida tropicalis and Candida glabrata after exposure to disinfectant solutions. METHODOLOGY: Simple biofilms were grown on heat-polymerized acrylic resin specimens, and divided into groups according to solutions/strains: distilled water (control); 0.25% sodium hypochlorite (NaOCl 0.25% ); 10% Ricinus communis (RC 10%); and 0.5% Chloramine T (CT 0.5%). The virulence factors were evaluated using the CFU count (microbial load), XTT method (cell metabolism), epifluorescence microscopy (biofilm removal and live or dead cells adhered), protease and phospholipase production and hyphae formation. Data were analyzed (α=0.05) by one-way ANOVA/ Tukey post hoc test, Kruskal-Wallis test and Wilcoxon test. RESULTS: NaOCl 0.25% was the most effective solution. CT 0.5% reduced the number of CFUs more than RC 10% and the control. RC 10% was effective only against C. glabrata. RC 10% and CT 0.5% decreased the cellular metabolism of C. albicans and C. glabrata. Enzyme production was not affected. Hyphal growth in the RC 10% and CT 0.5% groups was similar to that of the control. CT 0.5% was better than RC 10% against C. albicans and C. tropicalis when measuring the total amount of biofilm and number of living cells. For C. glabrata, CT 0.5% was equal to RC 10% in the maintenance of living cells; RC 10% was superior for biofilm removal. CONCLUSIONS: The CT 0.5% achieved better results than those of Ricinus communis at 10%, favoring the creation of specific products for dentures. Adjustments in the formulations of RC 10% are necessary due to efficacy against C. glabrata. The NaOCl 0.25% is the most effective and could be suitable for use as a positive control.

Expression of virulence factors by Pseudomonas aeruginosa biofilm after bacteriophage infection.

The use of bacteriophages for the treatment of bacterial infections has been extensively studied. Nonetheless, the stress response regarding bacteriophage infection and the expression of virulence factors of Pseudomonas aeruginosa after phage infection is poorly discussed. In this study, we evaluated biofilm formation capacity and expression of virulence factors of P. aeruginosa after bacteriophage infection. Biofilm growth rates, biofilm morphology, pyocyanin production and elastase activity were evaluated after 2, 8, 24 and 48 h of co-cultivation with bacteriophages that was recently characterized and showed to be infective towards clinical isolates. In parallel, quantitative real-time polymerase chain reactions were carried out to verify the expression of virulence-related genes. Bacteriophages promoted substantial changes in P. aeruginosa biofilm growth at early co-culture time. In addition, at 8 h, we observed that some cultures developed filaments. Although bacteriophages did not alter both pyocyanin and protease activity, changes on the expression level of genes related to virulence factors were detected. Usually, lasI, pslA, lasB and phzH genes were upregulated after 2 and 48 h of co-culture. These results highlight the need for extensive investigation of pathways and molecules involved in phage infection, since the transcriptional changes would suggest a response activation by P. aeruginosa.

The Impact of an Ocular Prosthesis on the Quality of Life, Perceived Stress, and Clinical Adaptation of Anophthalmic Patients: A Clinical and Longitudinal Trial.

PURPOSE: This prospective study evaluated and correlated the impact of ocular prostheses on quality of life and stress with socioeconomic level and clinical characteristics. The clinical difficulties and perceptions of patients after ocular rehabilitation were evaluated. MATERIALS AND METHODS: Anophthalmic patients (at least 18 years of age) who were not users of ocular prostheses were recruited. The Medical Outcomes Short-Form Health Survey (SF-36) and Perceived Stress Scale (PSS-10) questionnaires were administered before and after 3 and 6 months of prosthesis installation. Clinical characteristics, difficulties, and perceptions were evaluated by quantitative and descriptive analysis (7 days, 3 months, 6 months). Data were analyzed by the Friedman test and Pearson Correlation test (α = 0.05). RESULTS: The final sample consisted of 26 patients. Quality of life showed improvement in the "Bodily Pain" and "General Health" domains. Wearing the prosthesis did not influence perceived stress. The clinical evaluation showed clinical discharge over 6 months and presence of pain only at 7 days. A weak correlation occurred between sociodemographic characteristics and the categories "Role-Physical" (r = 0.423) and "General Health" (r = 0.494); cause of anophthalmia and "Role-Physical" (r = -0.471); and type of surgery and "General Health" (r = -0.432). CONCLUSIONS: According to the results of this study, the provision of ocular prostheses showed positive influence in 2 domains of quality of life and weak correlations with socioeconomic level, type of surgery, and cause of loss. Ocular rehabilitation did not influence stress. The use and care of the prostheses did not affect tissue inflammation, but the discharge was continuous.

Effectiveness of Brushing Associated With Oral Irrigation in Maintenance of Peri-Implant Tissues and Overdentures: Clinical Parameters and Patient Satisfaction.

Hygiene of implant-supported prostheses and the health of peri-implant tissues are considered to be predictive factors of successful prosthetic rehabilitation. Therefore, the purpose of this study was to evaluate the effectiveness of brushing associated with oral irrigation measured as biofilm-removing capacity, maintenance of healthy oral tissues, and patient satisfaction. A randomized, crossover clinical trial was conducted with 38 patients who wore conventional maxillary complete dentures and mandibular overdentures retained by the O-ring system. The patients were instructed to use the following hygiene methods for 14 days: mechanical brushing (MB; brush and dentifrice, control) and MB with oral irrigation (Waterpik [WP], experimental). Biofilm-removing capacity and maintenance of healthy oral tissues were evaluated by the modified plaque index (PI), gingival index (GI), probing depth (PD), and bleeding on probing index (BP) recorded at baseline and after each method. In parallel, patients answered a specific questionnaire using a visual analog scale after each hygiene method. Data were analyzed by Friedman and Wilcoxon tests (α = .05). The results showed significantly lower PI, GI, PD, and BP after application of the hygiene methods (P < .001) than those observed at baseline. However, no significant difference was found between MB and WP. The responses to the satisfaction questionnaire for both methods showed high mean values for all questions, with no statistically significant difference found between the answers given after the use of MB and WP (P > .05). The findings suggest that WP was effective in reducing PI, GI, PD, and BP indices and provided a high level of patient satisfaction.

Brushing associated with oral irrigation in maintaining implants and overdentures hygiene - a randomized clinical trial.

Evaluate, through a randomized clinical trial, the efficacy of brushing associated with oral irrigation in maintaining implant and overdenture hygiene. Thirty-eight participants, who had a clinically acceptable conventional maxillary complete denture and mandibular overdenture retained by either implants or mini-implants using an O-ring-retained system, were enrolled to participate in the study. They were instructed to use two different hygiene methods, in a random sequence for a period of 14 days, with a 7-day wash-out interposed period: (I) mechanical brushing (MB); (II) association of mechanical brushing with oral irrigation (WP). Biofilms from both subgingival sulci and overdentures were collected and processed by Checkerboard DNA-DNA hybridization method at baseline and after using the proposed hygiene protocols. Comparisons were performed using Wilcoxon test and Friedman test with Benjamini-Hochberg false discovery rate, followed by Conover post-hoc test (α = 0.05). In the subgingival sulci-related biofilm, a lower number of microbial cells were detected, after WP compared to the MB method (P < 0.001). The findings of overdenture-related biofilm suggest that both methods were similar (P = 0.607) being the identified microbiota qualitatively coincident after each method. Despite the number of microbial counts, it was concluded that the association of mechanical brushing with oral irrigation was more effective in reducing microorganisms in the subgingival sulci biofilm; however, the same outcome was not observed in the overdentures.

Action of disinfectant solutions on adaptive capacity and virulence factors of the Candida spp. biofilms formed on acrylic resin

Abstract Understanding the behavior of Candida spp. when exposed to denture disinfectants is essential to optimize their effectiveness. Changes in the virulence factors may cause increased resistance of Candida spp. to disinfectant agents. Objective To evaluate the microbial load, cellular metabolism, hydrolytic enzyme production, hyphae formation, live cell and biofilm quantification of Candida albicans, Candida tropicalis and Candida glabrata after exposure to disinfectant solutions. Methodology Simple biofilms were grown on heat-polymerized acrylic resin specimens, and divided into groups according to solutions/strains: distilled water (control); 0.25% sodium hypochlorite (NaOCl 0.25% ); 10% Ricinus communis (RC 10%); and 0.5% Chloramine T (CT 0.5%). The virulence factors were evaluated using the CFU count (microbial load), XTT method (cell metabolism), epifluorescence microscopy (biofilm removal and live or dead cells adhered), protease and phospholipase production and hyphae formation. Data were analyzed (α=0.05) by one-way ANOVA/ Tukey post hoc test, Kruskal-Wallis test and Wilcoxon test. Results NaOCl 0.25% was the most effective solution. CT 0.5% reduced the number of CFUs more than RC 10% and the control. RC 10% was effective only against C. glabrata. RC 10% and CT 0.5% decreased the cellular metabolism of C. albicans and C. glabrata. Enzyme production was not affected. Hyphal growth in the RC 10% and CT 0.5% groups was similar to that of the control. CT 0.5% was better than RC 10% against C. albicans and C. tropicalis when measuring the total amount of biofilm and number of living cells. For C. glabrata, CT 0.5% was equal to RC 10% in the maintenance of living cells; RC 10% was superior for biofilm removal. Conclusions The CT 0.5% achieved better results than those of Ricinus communis at 10%, favoring the creation of specific products for dentures. Adjustments in the formulations of RC 10% are necessary due to efficacy against C. glabrata. The NaOCl 0.25% is the most effective and could be suitable for use as a positive control.

Complete denture hygiene solutions: antibiofilm activity and effects on physical and mechanical properties of acrylic resin

Abstract Appropriated denture hygiene is a predictive factor for longevity of rehabilitation treatment and maintenance of the oral mucosal health. Although, disinfectant solutions are commonly used as denture cleansers, the impact of these solutions on acrylic resin-based dentures remain unclear. Objective To evaluate, in vitro, the antibiofilm activity of complete denture hygiene solutions and their effects on physical and mechanical properties of acrylic resin. Methodology For antibiofilm activity measurement acrylic resin specimens were contaminated with Candida albicans, Candida glabrata and Streptococcus mutans. After biofilm growth, the specimens were assigned to the hygiene solutions: Distilled water (Control); 0.2% Sodium hypochlorite (SH); Efferdent Power Clean Crystals (EPC) and 6.25% Ricinus communis (RC). The viability of microorganisms was evaluated by agar plate counts. In parallel, physical, and mechanical properties of the acrylic resin were evaluated after simulating a 5-year period of daily immersion in the previously mentioned solutions. The changes in surface roughness, color, microhardness, flexural strength, impact strength, sorption and solubility were evaluated. Data were compared by ANOVA followed by the Tukey test or Kruskal-Wallis followed by the Dunn test depending on the distribution (α=0.05). Results Regarding antibiofilm action, SH eliminated all microorganisms while EPC and RC exhibited moderate action against S. mutans (p=0.001) and C. glabrata (p<0.001), respectively. Relative to effects on the physical and mechanical properties of the acrylic resin, RC led to higher values of color change (p=0.030), hardness (p<0.001), surface roughness (p=0.006) and flexural strength (p<0.001). Moreover, RC induced the highest values of changes in solubility (p<0.001). EPC promoted greater changes in surface morphology, whereas immersion in SH retained the initial appearance of the acrylic resin surface. All hygiene solutions reduced the impact strength (p<0.05). Conclusion SH presented the most effective antibiofilm activity. In addition, changes on properties were observed after immersion in RC, which were considered within acceptable limits.

The effects of three disinfection protocols on Candida spp., denture stomatitis, and biofilm: A parallel group randomized controlled trial.

STATEMENT OF PROBLEM: Antifungals are used to treat Candida infections. However, because of increased antifungal resistance and the length of antifungal therapy, Candida spp. infections can be prevented using the prosthesis hygiene method. Therefore, establishing efficient, safe, and low-cost hygiene protocols for complete denture wearers is necessary. PURPOSE: The purpose of this clinical trial was to compare 10% Ricinus communis (RC10%) and 0.5% chloramine-T (CT0.5%) with negative (water) and positive (0.25% sodium hypochlorite [SH0.25%]) controls to establish a protocol to treat denture stomatitis (DS), remove denture biofilm, reduce overall microbiota, and decrease Candida spp. on the palate and denture bases. MATERIAL AND METHODS: This randomized, double blind, controlled clinical trial allocated 60 DS-positive participants in parallel groups: RC10%, CT0.5%, negative control, and SH0.25%. All participants brushed their palate and dentures and applied 1 of the solutions only to the denture. The following outcomes were assessed at baseline and after 7 and 37 days: Candida spp. counts, frequency of species by presumptive identification, DS severity, and photographic quantification of biofilm. The Kruskal-Wallis and Friedman tests with stepwise step-down post hoc test compared the anticandidal effect and the DS score (between groups and time). ANOVA and the Tukey post hoc test were used for biofilm removal comparison (α=.05). RESULTS: Microbial counts were solution- and time-dependent for dentures, with C. albicans, C. tropicalis, and C. glabrata being the most prevalent species. RC10% presented similar results to baseline and control after 7 and 37 days. CT0.5% reduced the CFU/mL compared with the baseline. SH0.25% was the most effective. DS reduced in all groups, independent of the solution. SH0.25% reduced biofilm the most, followed by RC10%. CT0.5% was similar to the control. CONCLUSIONS: SH0.25% demonstrated potential for Candida spp. control in denture wearers with DS. The other protocols showed intermediate activity and might be more suitable for longer immersion periods.

Antibiofilm Activity of an Experimental Ricinus Communis Dentifrice on Soft Denture Liners.

The disadvantage of liners materials is the difficulty of biofilm control. It was compared an experimental dentifrice contained Ricinus communis, with commercials dentifrices as antibiofilm activity against microorganisms on denture liner. Six hundred specimens were distributed in 5 groups (n=18/ microorganism): water; experimental dentifrice; specific dentifrice for denture and two conventional dentifrices against C. albicans; C. glabrata; S. mutans; S. aureus; E. coli. Each group had a negative (n=5; without contamination) and positive control (n=15/ microorganism; without cleaning). The antibiofilm activity was evaluated by the method of biofilm formation in triplicate. The specimens were contaminated in a standard way and incubated. After that, manual brushing was performed (60 s), washed with PBS, immersed in liquid culture medium for resuspension and sowing in solid medium. The results (mean of triplicates) were expressed in CFU/mL. The data was submitted to Shapiro-Wilk, ANOVA and Tukey test (p<0.05). The specific dentifrice (1.27±1.20) was the most effective against S. mutans, followed by conventional (Trihydral, 3.13±0.88; Colgate, 2.16±2.02) and experimental (3.81±1.37) dentifrices, which were similar to each other (p=0.008). All of them were different from water (4.79±1.42). The specific (0.21±0.21) and experimental (0.36±0.25) dentifrices were similar against S. aureus, with a higher mean of CFU when compared to conventional (Colgate, 0.06±0.13), which was more efficient (p=0.000). For C. albicans, C. glabrata and E. coli, all dentifrices were similar to water (p=0.186). It was concluded, that the experimental dentifrice was effective against S. aureus and had not efficacy against Candida spp.; S. mutans; E. coli, as occurred with the commercials dentifrices.

In Vitro Evaluation of Resilient Liner after Brushing with Conventional and Experimental Ricinus communis-Based Dentifrices.

PURPOSE: To evaluate the effect of experimental (Ricinus communis) and commercial dentifrices used for denture cleaning on abrasiveness (gravimetric method; roughness), hardness, and color stability of a resilient relining material. MATERIALS AND METHODS: Sixty circular (15 × 3 mm) specimens were distributed into four groups: C (control; brushing with water); CO (brushing with Colgate - for natural teeth); CB (brushing with Corega Brite - for complete dentures); RC (brushing with experimental dentifrice). Brushing was performed in a toothbrushing machine with a soft brush and a dentifrice suspension for 50 minutes, calculated to correspond to 1 year of regular brushing. Variables were measured initially and after the trial period. For the gravimetric method, the difference in mass was considered. The surface roughness was measured by a rugosimeter, and the hardness test was performed by a Shore A durometer. Color changes (ΔE; CIE L*a*b* and NBS systems) were measured by a portable spectrophotometer. Results were analyzed by ANOVA/Tukey's test (α = 0.05). RESULTS: The largest mass variation (µg; p < 0.0001) occurred in C (-6.21 ± 3.18). Concerning roughness, CB (0.26 ± 0.04) showed the lowest value, followed by RC (0.29 ± 0.08) and CO (0.34 ± 0.24) (p < 0.0001). Group C produced the greatest surface roughness (0.72 ± 0.25). Hardness values decreased after brushing with water (p = 0.014). No significant differences were found among RC (50.31 ± 1.03), CO (49.11 ± 1.31), CB (49.17 ± 1.23), and C (48.02 ± 1.26). Color stability was similar in all groups (p = 0.135; C: 2.3 ± 0.77; CO: 2.6 ± 0.54; CB: 2.2 ± 0.44; RC: 2.9 ± 1.56). CONCLUSIONS: The use of experimental dentifrice could be indicated, as it showed similar results to the specific dentifrice, keeping the resilient material properties within acceptable values.

The Effect of a Continuous Mechanical Polishing Protocol on Surface Roughness, Biofilm Adhesion, and Color Stability of Acrylic Resin Artificial Teeth.

PURPOSE: Acrylic resin properties are susceptible to change over the duration of use, thereby decreasing prosthesis longevity. To make the material less susceptible to the action of external agents such as stain-causing liquids and cleaning solutions, polishing procedures are recommended. A specific mechanical polishing procedure performed regularly was evaluated regarding the biofilm adhesion, surface roughness (Ra), color stability (ΔE), and mass changes in acrylic resin artificial teeth. MATERIALS AND METHODS: Sixty artificial teeth divided into two groups (n = 30) were immersed in distilled water (WT) and coffee (CF). These groups were then subdivided (n = 10) according to the type of polishing procedure administered: no polishing, biweekly polishing (once every 2 weeks) (pol 1), or monthly polishing (once a month) (pol 2), using aluminum oxide paste and a felt polishing wheel attached to an electric motor at a speed of 3000 rpm. Properties were evaluated at baseline and after 4, 8, 12, and 24 months of simulated immersion. At the end, the adhesion of Candida albicans and Streptococcus mutans multispecies biofilm in all groups was assessed. RESULTS: The CF group showed an increased Ra and weight, and a significantly greater ΔE compared to the WT group. The polishing procedure reduced Ra and minimized the stains caused by coffee, without losing mass, with the biweekly regime more effective than the monthly regime. Also, polishing reduced the adhesion of biofilm in the CF groups, again with the biweekly regime the most effective. CONCLUSIONS: When performed regularly, the mechanical polishing procedure tested reduces the changes in artificial teeth subjected to immersion in coffee, with the biweekly frequency most effective regarding the properties evaluated.