Effect of effervescent tablets on removable partial denture hygiene.

PURPOSE: To evaluate the effectiveness of five alkaline peroxide-based effervescent tablets in reducing both biofilms and the food layer adhered on the cobalt-chromium surface. METHODS: Cobalt-chromium metal alloy specimens were contaminated with Candida albicans, Candida glabrata, Streptococcus mutans and Staphylococcus aureus. After biofilm maturation, the specimens were immersed in Polident 3 Minute, Polident for Partials, Efferdent, Steradent, Corega Tabs or distilled water (control). Residual biofilm rates were determined by colony forming units counts and biofilm biomass. In parallel, to investigate the denture cleaning capability of effervescent tablets, artificially contaminated removable partial dentures were treated with each cleanser. Data were analyzed by Kruskal-Wallis followed by Dunn post hoc test or ANOVA followed by Tukey post hoc test (α= 0.05). RESULTS: None of the hygiene solutions reduced C. albicans biofilm. Efferdent and Corega Tabs promoted reduction of C. glabrata biofilm, while Steradent was favorable against S. aureus biofilm. For S. mutans, lower biofilm rates were observed after immersion in Polident for Partials and Steradent. The effervescent tablets showed good cleaning performance, removing an artificial layer with carbohydrates, proteins, and fats, however, they were not effective in removing aggregated mature biofilm. CLINICAL SIGNIFICANCE: The different effervescent tablets presented favorable antimicrobial activity against C. glabrata, S. mutans and S. aureus on cobalt-chromium surfaces and showed cleaning capability. However, for an appropriate biofilm control, a complementary method should be evaluated since none of the peroxide-based solutions reduced C. albicans biofilms or substantially removed aggregated biofilm.

In situ evaluation of microbial profile formed on Ti-6Al-4V additive manufacturing disks: 16S rRNA sequencing.

STATEMENT OF PROBLEM: Dental implants obtained by additive manufacturing may present changes in the microbiome formed. However, studies profiling the microbial communities formed on Ti-6Al-4V are lacking. PURPOSE: The purpose of this in situ study was to characterize the profile of the microbial communities formed on Ti-6Al-4V disks produced by additive manufacturing and machining. MATERIAL AND METHODS: Titanium disks produced by additive manufacturing (AMD) and machining (UD) were housed in the buccal region of removable intraoral devices. These devices containing both disks were used by eight participants for 96 hours. After every 24 hours of intraoral exposure, the biofilm that had formed on the disks was collected. The 16S rRNA genes from each specimen were amplified and sequenced with the Miseq Illumina instrument and analyzed. Total microbial quantification was evaluated by analysis of variance-type statistics using the nparLD package. The Wilcoxon test was used to evaluate alpha diversity (α=.05). RESULTS: A difference was found in the microbial communities formed on additively manufactured and machined disks, with a reduction in operational taxonomic units (OTUs) for the AMD group compared with the UD group. Firmicutes and Proteobacteria were the most abundant phyla. Of the 1256 genera sequenced, Streptococcus predominated on both disks. CONCLUSIONS: The microbiome of the biofilm formed on the Ti-6Al-4V disks was significantly influenced by the fabrication method. The AMD disks showed lower total microbial counts than the UD disks.

Effect of local hygiene protocols on denture-related stomatitis, biofilm, microbial load, and odor: A randomized controlled trial.

STATEMENT OF PROBLEM: Denture stomatitis affects complete denture wearers and is frequently treated with antifungals drugs, as well as treating the denture with sodium hypochlorite. Whether the limitations of these treatments can be overcome with local hygiene protocols that do not damage the denture materials or adversely affect the patient is unclear. PURPOSE: The purpose of this randomized controlled trial was to evaluate the effect of denture hygiene protocols on complete denture wearers with denture stomatitis. MATERIAL AND METHODS: For this randomized, double-blind controlled clinical trial, 108 participants were assigned to parallel groups: 0.25% sodium hypochlorite (positive control) 0.15% Triclosan, denture cleaning tablets, or denture cleaning tablets plus gingival cleaning tablets. The participants were instructed to brush the dentures and the palate and immerse the denture in the solutions. The outcomes of denture stomatitis remission, biofilm removal, decrease of microbial load (colony-forming units), and odor level of the mouth and denture were measured at baseline and after 10 days. Descriptive analyses were used for sociodemographic characterization of the participants; the Pearson chi-square test was used to compare participant frequency with different degrees of denture stomatitis. The data were not normally distributed (Shapiro-Wilks test) or homogeneous (Levene test). So, the Kruskal-Wallis and Dunn post hoc tests and Wilcoxon test were used to compare the effects of solutions and time on the variables (α=.05). RESULTS: The frequency of the highest to lowest denture stomatitis scores was significantly different for the 0.15% Triclosan and denture cleaning tablets groups. No significant difference was found among the groups in terms of denture stomatitis scores, biofilm, or colony-forming unit count of Candida spp. or C. albicans and S. mutans; a significant reduction was found in these parameters. The 0.25% sodium hypochlorite and 0.15% Triclosan treatments caused a significant reduction in Gram-negative microorganisms; these 2 protocols, and the denture cleaning tablets showed a significant reduction in Staphylococcus spp.; all protocols had similar effects. Only the S. mutans count of the palate decreased after 10 days. The odor level of the mouth and the denture was not significantly different (P=.778). CONCLUSIONS: The evaluated protocols can be recommended for the hygiene of complete dentures, since they were effective for all the variables studied.

Effectiveness of Oil-Based Denture Dentifrices-Organoleptic Characteristics, Physicochemical Properties and Antimicrobial Action.

Denture dentifrices must be effective and not deleterious to prosthetic devices. This study formulated and evaluated dentifrices based on oils of Copaifera officinalis, Eucalyptus citriodora, Melaleuca alternifolia, Pinus strobus, and Ricinus communis. Organoleptic characteristics (appearance, color, odor, taste), physicochemical properties (pH, density, consistency, rheological, abrasiveness, weight loss, and surface roughness) and antimicrobial (Hole-Plate Diffusion-HPD)/anti-biofilm (Colony Forming Units-CFU) action against Staphylococcus aureus, Streptococcus mutans, and Candida albicans were evaluated. Formulations were compared with water (negative control) and a commercial dentifrice (positive control). The data were analyzed by Kruskal-Wallis and Dunn tests (α = 0.05). The organoleptic and physicochemical properties were adequate. All dentifrices promoted weight losses, with high values for C. officinalis and R. communis, and an increase in surface roughness, without differing from each other. For antimicrobial action, C. officinalis and E. citriodora dentifrices were similar to positive control showing effectiveness against S. mutans and C. albicans and no dentifrice was effective against S. aureus; regarding the anti-biofilm action, the dentifrices were not effective, showing higher CFU counts than positive control for all microorganisms. The dentifrices presented satisfactory properties; and, although they showed antimicrobial action when evaluated by HPD, they showed no effective anti-biofilm action on multispecies biofilm.

Bacteriophage Cocktail-Mediated Inhibition of Pseudomonas aeruginosa Biofilm on Endotracheal Tube Surface.

Although different strategies to control biofilm formation on endotracheal tubes have been proposed, there are scarce scientific data on applying phages for both removing and preventing Pseudomonas aeruginosa biofilms on the device surface. Here, the anti-biofilm capacity of five bacteriophages was evaluated by a high content screening assay. We observed that biofilms were significantly reduced after phage treatment, especially in multidrug-resistant strains. Considering the anti-biofilm screens, two phages were selected as cocktail components, and the cocktail's ability to prevent colonization of the endotracheal tube surface was tested in a dynamic biofilm model. Phage-coated tubes were challenged with different P. aeruginosa strains. The biofilm growth was monitored from 24 to 168 h by colony forming unit counting, metabolic activity assessment, and biofilm morphology observation. The phage cocktail promoted differences of bacterial colonization; nonetheless, the action was strain dependent. Phage cocktail coating did not promote substantial changes in metabolic activity. Scanning electron microscopy revealed a higher concentration of biofilm cells in control, while tower-like structures could be observed on phage cocktail-coated tubes. These results demonstrate that with the development of new coating strategies, phage therapy has potential in controlling the endotracheal tube-associated biofilm.

Antimicrobial activity of effervescent denture tablets on multispecies biofilms.

PURPOSE: To investigate the impact of peroxide-based solutions in reducing viability and metabolic activity of multispecies biofilms on denture base acrylic resin surfaces and for removing them from these surfaces. BACKGROUND: Denture cleansers are effective in reducing monospecies biofilm; however, studies evaluating their action on multispecies biofilms are scarce. MATERIALS AND METHODS: Sixty-nine denture base acrylic resin specimens (Ø 15 × 3 mm) were sterilised then contaminated with Candida albicans, Staphylococcus aureus and Pseudomonas aeruginosa to form multispecies biofilms. Biofilms were grown for 24 hours; subsequently, specimens were immersed in three different cleansing solutions (n = 9): nitradine (NI), fixodent (FX) and phosphate-buffered saline (Control), according to the respective manufacturer's instructions. After applying the hygiene protocols, viability of microorganisms was evaluated by counting colony-forming units and assessing metabolic activity. Moreover, biofilm removal capacity was estimated based on extension of cell-covered areas visualised in fluorescent microscopy micrographics. RESULTS: Microbial counts were solution-dependent; NI was effective against all microorganisms (P < .05). FX exhibited moderate antimicrobial action, reducing P aeruginosa (P < .05) and S aureus (P < .05) viability by approximately 2 logs. Both peroxide-based solutions reduced metabolic activity (P < .001) and biofilm-covered areas on specimen surfaces (P < .001). CONCLUSION: Under the experimental conditions tested, these results demonstrated that peroxide-based solutions had favourable antimicrobial activity but promoted no broad elimination of aggregated multispecies biofilm. NI might be more suitable as complementary chemical agent for controlling multispecies denture biofilm.

Identification and Characterization of New Bacteriophages to Control Multidrug-Resistant Pseudomonas aeruginosa Biofilm on Endotracheal Tubes.

Studies involving antimicrobial-coated endotracheal tubes are scarce, and new approaches to control multidrug-resistant Pseudomonas aeruginosa biofilm on these devices should be investigated. In this study, five new P. aeruginosa bacteriophages from domestic sewage were isolated. All of them belong to the order Caudovirales, Myoviridae family. They are pH and heat stable and produce 27 to 46 particles after a latent period of 30 min at 37°C. Their dsDNA genome (ranging from ∼62 to ∼65 kb) encodes 65 to 89 different putative proteins. They exhibit a broad lytic spectrum and infect 69.7% of the P. aeruginosa strains tested. All the bacteriophages were able to reduce the growth of P. aeruginosa strains in planktonic form. The bacteriophages were also able to reduce the biofilm viability rates and the metabolic activity of P. aeruginosa strains in a model of biofilms associated with endotracheal tubes. In addition, scanning electron microscopy micrographs showed disrupted biofilms and cell debris after treatment of bacteriophages, revealing remarkable biofilm reduction. The lytic activity on multidrug-resistant P. aeruginosa biofilm indicates that the isolated bacteriophages might be considered as good candidates for therapeutic studies and for the application of bacteriophage-encoded products.

Hygiene protocols for the treatment of denture-related stomatitis: local and systemic parameters analysis - a randomized, double-blind trial protocol.

BACKGROUND: Denture-related stomatitis (DS) is chronic multifactorial inflammation, strongly related to the presence of the biofilm that is the complex structure formed by microorganisms held together by a mucus-like matrix of carbohydrate that adheres to different surfaces, including the denture surface. DS has recently been correlated with deleterious cardiovascular alterations. The potential effect of hygiene protocols in the control of DS and randomized clinical trials that address this oral condition with cardiovascular complications are important in clinical decision-making. MATERIAL/DESIGN: A clinical trial, randomized, double-blind, and with parallel groups, will be conducted in Brazil The sample will consist of 100 patients without teeth in both arches, users of at least maxillary complete dentures, and diagnosed with DS, who will be allocated to groups (n = 25 per group) according to the different hygiene protocols: (1) brushing of the palate and immersion of the prosthesis in 0.25% sodium hypochlorite solution (positive control); (2) brushing of the palate and immersion of the prosthesis in 0.15% triclosan solution; (3) brushing of the palate and immersion of the prosthesis in lactose monohydrate; or (4) brushing the palate with citric acid and immersing the prosthesis in lactose monohydrate. The response variables will be heart rate variability and alteration of blood pressure (systemic level), remission of DS, removal of biofilm, reduction of microbial load (colony-forming units (CFU)), mouth and prosthesis odor level, expression of MUC1, proinflammatory cytokines, C-reactive protein (CRP), viscosity, pH and salivary flow (locally); patient-centred qualitative analysis will also be undertaken. Measurements will be performed at baseline and 10 days after the interventions. The results obtained will be statistically analyzed as pertinent, with a level of significance of 0.05. DISCUSSION: This study will provide a guideline for clinical practice regarding the use of hygiene protocols in the treatment of oral diseases (DS) mediated by biofilm. Also, it may provide evidence of correlation of oral manifestation with cardiac risk. TRIAL REGISTRATION: Brazilian Registry of Clinical Trials, RBR-4hhwjb. Registered on 9 November 2018.

Influence of AgVO3 incorporation on antimicrobial properties, hardness, roughness and adhesion of a soft denture liner.

The objective of this in vitro study was to investigate the effect of nanostructured silver vanadate decorated with silver nanoparticles (AgVO3) on antimicrobial activity, hardness, roughness, and adhesion of a soft denture liner. The antimicrobial efficacy of the Trusoft (Boswoth) liner incorporated with different concentrations of AgVO3 against Enterococcus faecalis, Pseudomonas aeruginosa, Candida albicans, and Staphyloccocus aureus (n = 5) was evaluated by the agar diffusion method. Roughness, hardness, and adhesion properties were also evaluated. The data were analyzed by analysis of variance (ANOVA) and Tukey's multiple comparison test with significance at the p < 0.05 level. At concentrations of 1 and 2.5%, AgVO3 incorporation was effective only against E. faecalis, and at 5 and 10%, against E. faecalis, P. aeruginosa, and C. albicans. None of the concentrations was effective against S. aureus. A decrease in hardness was found for the 1, 2.5, and 10% AgVO3 concentrations (p < 0.001) and at 5%, hardness was not affected. None of the concentrations affected the roughness of the material. A significant increase in tensile values was observed between the liner and heat-curing acrylic resin for 2.5% (p < 0.001) and 10% (p = 0.042) concentrations. AgVO3 incorporation to a soft denture liner promoted antimicrobial activity against E. faecalis, P. aeruginosa, and C. albicans without affecting roughness, maintaining the hardness properties recommended for soft and extra soft liners, and improving the adhesion between the liner and the acrylic resin used for dentures.

Development of an Impression Material with Antimicrobial Properties for Dental Application.

PURPOSE: To evaluate the antimicrobial activity and physico-mechanical properties of an irreversible hydrocolloid in which nanostructured silver vanadate decorated with silver nanoparticles (AgVO3 ) was added at various concentrations (0% - control, 2.5%, 5%, and 10% by weight). MATERIALS AND METHODS: The agar diffusion method (n = 10) was used to evaluate the inhibitory effect on the following species: Streptococcus mutans, Staphyloccocus aureus, Pseudomonas aeruginosa, Escherichia coli, and Candida albicans. The gelation time, flow capacity and plastic deformation were verified (n = 10). The data were analyzed using the Kruskal-Wallis test followed by the Dunn post-test, or via one-way ANOVA with multiple comparisons with a Bonferroni adjustment depending on the distribution (α = 0.05). RESULTS: All percentages of the nanomaterial were able to promote the antimicrobial activity of a hydrocolloid, with the formation of an inhibition zone (p < 0.05). In general, there was a dose-dependent effect on antimicrobial activity: higher concentrations of the nanomaterial promoted greater action except in the cases of P. aeruginosa (p < 0.001; F = 51.74) and S. aureus (p < 0.001), where the highest inhibition was for the 2.5% group. No difference was found in the gelation time when the control was compared with the groups with AgVO3 (p > 0.05). The difference was between the 5% and 10% groups (p = 0.007), and the latter promoted an increase in time. The flow capacity of the hydrocolloid with 5% of AgVO3 was significantly lower when compared with the control (p = 0.034). The AgVO3 influenced the plastic deformation (p < 0.001) in such a way that concentrations of 5% (p = 0.010) and 10% (p < 0.001) promoted an increase in this property when compared with the control. CONCLUSIONS: AgVO3 can be incorporated into an irreversible hydrocolloid as an antimicrobial agent without promoting adverse effects on physical-mechanical properties when used in concentrations of 2.5%.

Evaluation of biofilm removal and adverse effects on acrylic resin by diluted concentrations of sodium hypochlorite and Ricinus communis solutions.

PURPOSE: To verify whether 0.1% and 0.2% sodium hypochlorite (NaOCl), and 8% Ricinus communis (RC) were able to remove denture biofilm without causing deleterious effects to acrylic resin. BACKGROUND: Previous data show that denture cleansers are effective in reducing biofilm; however, they can change acrylic resin properties. METHODS: In a crossover trial, 47 denture wearers brushed and soaked their dentures (20 min/14 d): control, 0.85% saline; SH1, 0.1% NaOCl; SH2, 0.2% NaOCl and RC. Denture biofilm on the intaglio surface was stained, photographed and quantified (Image Tool® ). Furthermore, 80 rectangular and 80 disc-shaped specimens (Lucitone 550) were assigned into tested solutions (n = 20), simulating 5 years of daily short immersions (20 minutes). A colorimeter and the National Bureau of Standards units (NBS) determined colour data (ΔE). Surface roughness and flexural strength were measured using rugosimeter and universal testing machine, respectively. Data were compared by the Friedman test (α = .05) followed by Wilcoxon, corrected by Bonferroni (α = .005) (clinical) and Kruskal-Wallis followed by the Dunn test (α = .05) (laboratorial). RESULTS: SH2 (MR=1.77) showed lower biofilm coverage; SH1 (MR = 2.37) and RC (MR = 2.74) presented intermediated values. RC (1.10 [0.96:1.75]) revealed higher colour alteration than SH1 (0.71 [0.62:0.80]) and SH2 (0.74 [0.58:0.85]); however, NBS classified all solutions as "trace" (0.0-0.5). There was no statistical significance for surface roughness (P = .760) and flexural strength (P = .547). CONCLUSIONS: The 0.2% NaOCl showed the best clinical performance and did not cause adverse effects on acrylic resin on laboratory analyses.

Growth and functional harvesting of human mesenchymal stromal cells cultured on a microcarrier-based system.

Human mesenchymal stromal cells (hMSCs) cells are attractive for applications in tissue engineering and cell therapy. Because of the low availability of hMSCs in tissues and the high doses of hMSCs necessary for infusion, scalable and cost-effective technologies for in vitro cell expansion are needed to produce MSCs while maintaining their functional, immunophenotypic and cytogenetic characteristics. Microcarrier-based culture systems are a good alternative to traditional systems for hMSC expansion. The aim of the present study was to develop a scalable bioprocess for the expansion of human bone marrow mesenchymal stromal cells (hBM-MSCs) on microcarriers to optimize growth and functional harvesting. In general, the results obtained demonstrated the feasibility of expanding hBM-MSCs using microcarrier technology. The maximum cell concentration (n = 5) was ~4.82 ± 1.18 × 10(5) cell mL(-1) at day 7, representing a 3.9-fold increase relative to the amount of inoculated cells. At the end of culture, 87.2% of the cells could be harvested (viability = 95%). Cell metabolism analysis revealed that there was no depletion of important nutrients such as glucose and glutamine during culture, and neither lactate nor ammonia byproducts were formed at inhibitory concentrations. The cells that were recovered after the expansion retained their immunophenotypic and functional characteristics. These results represent an important step toward the implementation of a GMP-compliant large-scale production system for hMSCs for cellular therapy.

Mycobacterium leprae intracellular survival relies on cholesterol accumulation in infected macrophages: a potential target for new drugs for leprosy treatment.

We recently showed that Mycobacterium leprae (ML) is able to induce lipid droplet formation in infected macrophages. We herein confirm that cholesterol (Cho) is one of the host lipid molecules that accumulate in ML-infected macrophages and investigate the effects of ML on cellular Cho metabolism responsible for its accumulation. The expression levels of LDL receptors (LDL-R, CD36, SRA-1, SR-B1, and LRP-1) and enzymes involved in Cho biosynthesis were investigated by qRT-PCR and/or Western blot and shown to be higher in lepromatous leprosy (LL) tissues when compared to borderline tuberculoid (BT) lesions. Moreover, higher levels of the active form of the sterol regulatory element-binding protein (SREBP) transcriptional factors, key regulators of the biosynthesis and uptake of cellular Cho, were found in LL skin biopsies. Functional in vitro assays confirmed the higher capacity of ML-infected macrophages to synthesize Cho and sequester exogenous LDL-Cho. Notably, Cho colocalized to ML-containing phagosomes, and Cho metabolism impairment, through either de novo synthesis inhibition by statins or depletion of exogenous Cho, decreased intracellular bacterial survival. These findings highlight the importance of metabolic integration between the host and bacteria to leprosy pathophysiology, opening new avenues for novel therapeutic strategies to leprosy.