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Purpose: The aim of this randomized clinical trial was to compare the effectiveness of microwave denture disinfection and nystatin in the treatment of well-controlled type 2 diabetic patients with denture stomatitis in terms of microbiologic and clinical outcomes. Materials and Method: Diabetic patients wearing maxillary complete dentures with denture stomatitis (n = 40) were divided into two groups: NYS (patients treated with topical nystatin 4 times/day for 14 days) and MW (patients who had their dentures microwaved [650 W for 3 minutes] 3 times/week for 14 days). Mycologic samples were taken from the palates and dentures of the patients for quantification and identification of Candida, and standardized photographs of the palates were taken for clinical analysis. Evaluations were repeated at baseline, the end of treatment (day 14), and throughout follow-up (days 30, 60, and 90). Microbiologic data were evaluated by analysis of variance using a random effects statistical model, Tukey post hoc test, and chi-square test ([alpha] = .05). Clinical resuts were analyzed using Mann-Whitney and Fisher exact tests ([alpha] = .05). Results: Both treatments were considered successful in reducing the clinical signs of denture stomatitis and significantly reduced the values of colony-forming units/mL from the palates and dentures at days 14 and 30. In addition, 40% of treated patients were cured by the end of treatment. No significant differences in the microbiologic and clinical outcomes were revealed between the two groups (P > .05). C albicans was the most predominant species isolated (P < .01), followed by C tropicalis and C glabrata. Conclusion: Denture microwave disinfection was as effective as nystatin for the treatment of diabetic patients with denture stomatitis. [ABSTRACT FROM AUTHOR]

Objectives: To evaluate the effect of microwave disinfection on the flexural strength and Vickers hardness of 4 autopolymerized resins (Kooliner [K], Tokuso Rebase Fast [T], Ufi Gel Hard [U], and New Truliner [N]) and 1 denture base resin (Lucitone 550 [L]). Method and Materials: For each material, 48 specimens (64 x 10 x 3.3 mm) were made and divided into 6 equal groups (n = 8). In the control group, specimens were untreated. Before testing, specimens were immersed in 200 mL of distilled water and submitted to disinfection for 1 of the following irradiation times: 1, 2, 3, 4, or 5 minutes. The irradiation procedure was performed twice. The flexural strength was determined using a testing machine MTS-810 and measurements of Vickers hardness were made on Micromet 2100. The values were submitted to ANOVA and Tukey's test (P = .05). Results: The K material showed a significant increase (P = .0010) in flexural strength following 5 minutes of disinfection compared to control specimens. The flexural strength mean values of materials T, U, and N were not significantly affected (P > .05) by disinfection. Compared to the control group, the K material showed a significant increase in hardness (P < .001) following disinfection for 3, 4, and 5 minutes. For material U, disinfection for 4 and 5 minutes produced specimens with significantly increased hardness values (P < .001) compared to the control group. For material N, disinfection for 5 minutes resulted in significantly higher hardness values (P < .001) than the control group. Conclusion: Regardless of the irradiation time, the flexural strength and hardness of the materials evaluated were not detrimentally affected by microwave disinfection. [ABSTRACT FROM AUTHOR]

PURPOSE: To evaluate the effect of 2 postpolymerization treatments on toothbrushing wear (weight loss) and surface roughness of 3 autopolymerized reline resins--Duraliner II (D) (Reliance Dental), Kooliner (K) (Coe Laboratories), and Tokuso Rebase Fast (T) (Tokuyama Dental)--and 1 heat-polymerized resin, Lucitone 550 (L) (Dentsply International). MATERIALS AND METHODS: Specimens (40 x 10 x 2mm) of each material (n = 24) were prepared and divided into 3 groups: control (no postpolymerization treatment); water bath (immersion in water at 55 degrees C); and microwave (microwave irradiation). Specimens were dried until constant weight was achieved and the surface roughness (Ra) was measured. Tests were performed in a toothbrush machine using 20,000 strokes of brushing at a weight of 200 g, with the specimens immersed in 1:1 dentifrice/water slurry. Specimens were reconditioned to constant weight and the weight loss (mg) and surface roughness were evaluated. Data were analyzed by 2-way analysis of variance and followed by Tukey test (alpha = .05). RESULTS: In the control group, the weight loss of materials D and T was lower (P < .05) than that of L. No differences among materials were found after postpolymerization treatments (P > .05). The weight loss of material T (control = 0.5 mg) was significantly increased (P < .05) after postpolymerization treatments (water bath = 1.9 mg; microwave = 1.8 mg). For materials K and T, the toothbrushed surface roughness was higher (P < .05) after microwave and waterbath postpolymerization treatments. Material L showed increased surface roughness after microwave postpolymerization treatment. CONCLUSION: The toothbrushing wear resistance of L was not superior to the reline resins. The postpolymerization treatments did not improve the toothbrushing wear resistance of the materials and produced an increased surface roughness for materials L, K, and T. [ABSTRACT FROM AUTHOR]

PURPOSE: The purpose of this study was to evaluate the effectiveness of microwave irradiation on the disinfection of simulated complete dentures. MATERIALS AND METHODS: Eighty dentures were fabricated in a standardized procedure and subjected to ethylene oxide sterilization. The dentures were individually inoculated (10(7) cfu/mL) with tryptic soy broth (TSB) media containing one of the tested microorganisms (Candida albicans, Streptoccus aureus, Bacillus subtilis, and Pseudomonas aeruginosa). After 48 hours of incubation at 37 degrees C, 40 dentures were individually immersed in 200 mL of water and submitted to microwave irradiation at 650 W for 6 minutes. Forty nonirradiated dentures were used as positive controls. Replicate aliquots (25 microL) of suspensions were plated at dilutions of 10(-3) to 10(-6) on plates of selective media appropriate for each organism. All plates were incubated at 37 degrees C for 48 hours. TSB beakers with the microwaved dentures were incubated at 37 degrees C for 7 more days. After incubation, the number of colony-forming units was counted and the data were statistically analyzed by Kruskal-Wallis test (alpha = .05). RESULTS: No evidence of growth was observed at 48 hours for S. aureus, B. subtilis, and C. albicans. Dentures contaminated with P. aeruginosa showed small growth on 2 plates. After 7 days incubation at 37 degrees C, no growth was visible in the TSB beakers of S. aureus and C. albicans. Turbidity was observed in 3 broth beakers, 2 from P. aeruginosa and 1 from B. subtilis. CONCLUSION: Microwave irradiation for 6 minutes at 650 W produced sterilization of complete dentures contaminated with S. aureus and C. albicans and disinfection of those contaminated with P. aeruginosa and B. subtilis. [ABSTRACT FROM AUTHOR]

Purpose: To evaluate the influence of water bath and microwave postpolymerization treatments on the cytotoxicity of 6 hard reline acrylic resins.Materials and Methods: The materials tested were Tokuso Rebase Fast (TR), Ufi Gel Hard (UGH), Duraliner II (D), Kooliner (K), New Truliner (NT), and Light Liner (LL). LL resin was additionally tested with an air-barrier coating (LLABC). Nine disks of each material (10 x 1 mm) were made and divided into 3 groups: group 1 (no postpolymerization treatment); group 2 (postpolymerization in microwave oven); group 3 (postpolymerization in water bath at 55 degrees C for 10 minutes). L929 cells were cultured in 96-well plates and incubated for 24 hours in Eagle's medium. Eluates prepared from the disks or medium without disks (control) replaced the medium. Cytotoxicity was assessed by both dehydrogenase succinic activity (MTT) assay and incorporation of radioactive 3H-thymidine assay. Tests were carried out in quadruplicate and repeated twice. Differences between groups were determined by analysis of variance with Tukey multiple-comparison intervals (alpha = .05).Results: For MTT assay, the postpolymerization treatments had no effect on the cytotoxicity of all materials (P > .05). For 3H-thymidine assay, the postpolymerization treatments significantly decreased the cytotoxicity of UGH (P < .05). The cytotoxicity of K, NT, LL, and LLABC increased after microwave irradiation (P < .05). TR, NT, and LLABC showed an increase in cytotoxicity after water bath (P < .05).Conclusion: When assessed by MTT assay, the cytotoxicity of the materials was not affected by postpolymerization treatments. 3H-Thymidine assay showed that the cytotoxicity of the resins was not improved by the postpolymerization treatments, with the exception of UGH. [ABSTRACT FROM AUTHOR]

The aims of this study were to evaluate the effects of Photogem®-mediated photosensitization on rat palatal mucosa and the biodistribution of the photosensitizer in this tissue. A solution of Photogem® (500 or 1000 mg/l) was applied to the palatal mucosa for 30 min and the exposure time to blue LED (460 nm) was 20 min (144 J/cm(2)). At 0, 1, 3, and 7 days, palatal mucosa was photographed for macroscopic analysis. After killing, the palate was removed for microscopic analysis. Thermal mapping evaluated temperature change in the tissue during irradiation. All experimental groups revealed intact mucosa in the macroscopic analysis. Tissue alterations were observed microscopically for only four out of 80 animals subjected to PDT. Fluorescence emitted by Photogem® was identified and was limited to the epithelial layer. A temperature increase from 35 to 41°C was recorded. Photogem®- mediated PDT was not toxic to the rat palatal mucosa. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the effectiveness of photodynamic therapy (PDT) for the disinfection of complete dentures. Biofilm samples were collected from dentures of 60 denture users who were randomly divided into four experimental groups (n = 15 each): subjects whose maxillary dentures were sprayed with 50 and 100 mg/l of Photogem® suspension (groups P50S and P100S) and patients whose maxillary dentures were treated with 50 and 100 mg/l of Photogem® gel (groups P50G and P100G). Dentures with photosensitizers were left in the dark for 30 min (pre-irradiation time) and then irradiated with blue LED light at 37.5 J/cm(2) (26 min). Denture samples were taken with sterile cotton swab before (left side surfaces) and after (right side surfaces) PDT. All microbial material was diluted and plated on selective media for Candida spp., Staphylococcus mutans spp., streptococci and a non-selective media. After incubation (48 h/37°C), the number of colony-forming units (cfu/ml) was counted. Microorganisms grown on selective media were identified using biochemical methods before and after PDT. The data were submitted to McNemar and Kruskal-Wallis tests (α = 0.05). No growth after PDT was observed in 60, 53, 47, and 40% of dentures from P100G, P50G, P100S, and P50S groups, respectively. When evidence of microorganisms' growth was observed, PDT regimens eliminated over 90% of microorganisms on dentures. This clinical study showed that PDT was effective for disinfecting dentures. [ABSTRACT FROM AUTHOR]

OBJECTIVE: Photodynamic therapy (PDT) is an effective method for Candida spp. inactivation in vitro and in vivo, but as yet, no clinical trial has been conducted. This report describes 5 cases of denture stomatitis (DS) treated with PDT. STUDY DESIGN: Five subjects with clinical and microbiologic diagnosis of DS were submitted to 6 sessions of PDT 3 times a week for 15 days. In each session, patients' dentures and palates were sprayed with 500 mg/L Photogem, and, after 30 minutes of incubation, irradiated by light-emitting diode light source at 455 nm (37.5 and 122 J/cm(2), respectively). Cultures of Candida spp. from dentures and palates and standard photographs of the palates were taken at baseline (day 0), at the end of the treatment (day 15), and at follow-up time intervals (days 30 and 60). RESULTS: Four patients showed clinical resolution of DS (no inflammation) after PDT sessions, and only 1 subject demonstrated reduction in palatal inflammation. Recurrence of DS was observed in 2 patients during the follow-up period. CONCLUSIONS: PDT appears to be an alternative treatment for DS. [ABSTRACT FROM AUTHOR]

STATEMENT OF PROBLEM: Microwave irradiation has been suggested as a method to disinfect denture bases. However, the effect of microwave heating on the dimensional stability of the relined denture bases is unknown. PURPOSE: The purpose of this study was to evaluate the dimensional stability of intact and relined acrylic resin denture bases after microwave disinfection. MATERIAL AND METHODS: A standard brass cast simulating an edentulous maxillary arch was machined and used to fabricate 2- and 4-mm-thick denture bases (n=200), which were processed with heat-polymerized acrylic resin (Lucitone 550). The 2-mm thick-specimens (n=160) were relined with 2 mm of autopolymerizing resin (Tokuso Rebase Fast, Ufi Gel Hard, Kooliner, or New Truliner). Distances between 5 removable pins on the standard brass cast were measured with a Nikon optical comparator, and the area (mm) formed by the distance between 5 pins was calculated and served as baseline. During fabrication, the pins were transferred to the intaglio surface of the specimens. Differences between the baseline area and those subsequently determined for the specimens were used to calculate the percent dimensional changes. The intact and relined denture bases were divided into 4 groups (n=10) and evaluated after: polymerization (control group P); 1 cycle of microwave disinfection (MW); daily microwave disinfection for 7 days (control group MW7); water storage for 7 days (WS7). Microwave irradiation was performed for 6 minutes at 650 W. Data were analyzed using 2-way ANOVA followed by Tukey's test (alpha=.05). RESULTS: Intact specimens and those relined with Kooliner and New Truliner showed increased shrinkage after 1 (P=.05, .018, and .001, respectively) and 7 (P <.001, .003, and <.001, respectively) cycles of microwave disinfection. With the exception of specimens relined with Kooliner, intact specimens showed greater shrinkage than the relined specimens after 7 cycles of microwave disinfection. CONCLUSIONS: Microwave disinfection produced increased shrinkage of intact specimens and those relined with New Truliner and Kooliner. [ABSTRACT FROM AUTHOR]

Objectives: To evaluate the effect of microwave postpolymerization heat treatment and water storage time on the cytotoxicity of denture base and reline acrylic resins. Method and Materials: Sample disks of 6 acrylic resins were fabricated under aseptic conditions (Kooliner, GC; Tokuyama Rebase II, Tokuyama Dental; New Truliner, Bosworth; Acron MC, GC; Lucitone 550, Dentsply; QC 20, Dentsply). The denture base acrylic resin samples were stored in water for 24 and 48 hours at 37°C. The fabricated samples were further subdivided into 2 groups: (1) samples without heat treatment and (2) samples treated in a microwave. The 3H-thymidine incorporation test was used to determine the cytotoxicity of the materials. The data were statistically analyzed using the incomplete 3-way ANOVA and Tukey HSD tests (P < .05). Results: Microwave postpolymerization heat treatment improved the biocompatibility of Tokuyama Rebase II. Tokuyama Rebase II without heat treatment and Acron MC in both experimental groups were graded as slightly cytotoxic in the group without water storage. The other resins were graded as noncytotoxic. After 24 hours of immersion in water, all materials were graded as noncytotoxic. After water storage for 48 hours, Acron MC without heat treatment and QC 20 in both experimental groups were graded as slightly cytotoxic. Lucitone 550 was graded as noncytotoxic in all experimental groups. Conclusion: Microwave irradiation may be considered an alternative to reduce the cytotoxicity of Tokuyama Rebase II. Dental practitioners should choose Lucitone 550 processed with terminal boiling stage. [ABSTRACT FROM AUTHOR]

Silver tungstate (α-Ag 2 WO 4 ), silver molybdate (β-Ag 2 MoO 4 ), and silver vanadate (α-AgVO 3 ) microcrystals have shown interesting antimicrobial properties. However, their biocompatibility is not yet fully understood. Cytotoxicity and the inflammatory response of silver-containing microcrystals were analyzed in THP-1 and THP-1 differentiated as macrophage-like cells, with the alamarBlue™ assay, flow cytometry, confocal microscopy, and ELISA. The present investigation also evaluated redox signaling and the production of cytokines (TNFα, IL-1β, IL-6, and IL-8) and matrix metalloproteinases (MMP-8 and -9). The results showed that α-AgVO 3 (3.9 μg/mL) did not affect cell viability ( p > 0.05). α-Ag 2 WO 4 (7.81 μg/mL), β-Ag 2 MoO 4 (15.62 μg/mL), and α-AgVO 3 (15.62 μg/mL) slightly decreased cell viability ( p ≤ 0.003). All silver-containing microcrystals induced the production of O 2 - and this effect was mitigated by Reactive Oxygen Species (ROS) scavenger and N-acetylcysteine (NAC). TNFα, IL-6 and IL-1β were not detected in THP-1 cells, while their production was either lower ( p ≤ 0.0321) or similar to the control group ( p ≥ 0.1048) for macrophage-like cells. The production of IL-8 by both cellular phenotypes was similar to the control group ( p ≥ 0.3570). The release of MMP-8 was not detected in any condition in THP-1 cells. Although MMP-9 was released by THP-1 cells exposed to α-AgVO 3 (3.9 μg/mL), no significant difference was found with control ( p = 0.7). Regarding macrophage-like cells, the release of MMP-8 and -9 decreased in the presence of all microcrystals ( p ≤ 0.010). Overall, the present work shows a promising biocompatibility profile of, α-Ag 2 WO 4 , β-Ag 2 MoO 4 , and α-AgVO 3 microcrystals., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Pimentel, De Annunzio, Assis, Barbugli, Longo and Vergani.)

Objective: Some microorganisms, i.e., Candida albicans, have been associated with cancer onset and development, although whether the fungus promotes cancer or whether cancer facilitates the growth of C. albicans is unclear. In this context, microbial-derived molecules can modulate the growth and resistance of cancer cells. This study isolated extracellular lipids (ECL) from a 36-h Candida albicans biofilm incubated with oral dysplastic (DOK) and neoplastic (SCC 25) cells, which were further challenged with the topoisomerase I inhibitor camptothecin (CPT), a lipophilic anti-tumoral molecule., Methodology: ECL were extracted from a 36-h Candida albicans biofilm with the methanol/chloroform precipitation method and identified with Nuclear Magnetic Resonance (1H-NMR). The MTT tetrazolium assay measured ECL cytotoxicity in DOK and SCC 25 cells, alamarBlue™ assessed cell metabolism, flow cytometry measured cell cycle, and confocal microscopy determined intracellular features., Results: Three major classes of ECL of C. albicans biofilm were found: phosphatidylinositol (PI), phosphatidylcholine (PC), and phosphatidylglycerol (PG). The ECL of C. albicans biofilm had no cytotoxic effect on neither cell after 24 hours, with a tendency to disturb the SCC 25 cell cycle profile (without statistical significance). The ECL-induced intracellular lipid droplet (LD) formation on both cell lines after 72 hours. In this context, ECL enhanced cell metabolism, decreased the response to CPT, and modified intracellular drug distribution., Conclusion: The ECL (PI, PC, and PG) of 36-h Candida albicans biofilm directly interacts with dysplastic and neoplastic oral cells, highlighting the relevance of better understanding C. albicans biofilm signaling in the microenvironment of tumor cells.

In this work, α-Ag 2-2 x Cu x WO 4 (0 ≤ x ≤ 0.16) solid solutions with enhanced antibacterial (against methicillin-resistant Staphylococcus aureus ) and antifungal (against Candida albicans ) activities are reported. A plethora of techniques (X-ray diffraction with Rietveld refinements, inductively coupled plasma atomic emission spectrometry, micro-Raman spectroscopy, attenuated total reflectance-Fourier transform infrared spectroscopy, field emission scanning electron microscopy, ultraviolet-visible spectroscopy, photoluminescence emissions, and X-ray photoelectron spectroscopy) were employed to characterize the as-synthetized samples and determine the local coordination geometry of Cu 2+ cations at the orthorhombic lattice. To find a correlation between morphology and biocide activity, the experimental results were sustained by first-principles calculations at the density functional theory level to decipher the cluster coordinations and electronic properties of the exposed surfaces. Based on the analysis of the under-coordinated Ag and Cu clusters at the (010) and (101) exposed surfaces, we propose a mechanism to explain the biocide activity of these solid solutions.

In the current COVID-19 pandemic, the next generation of innovative materials with enhanced anti-SARS-CoV-2 activity is urgently needed to prevent the spread of this virus within the community. Herein, we report the synthesis of chitosan/α-Ag 2 WO 4 composites synthetized by femtosecond laser irradiation. The antimicrobial activity against Escherichia coli, Methicilin-susceptible Staphylococcus aureus (MSSA), and Candida albicans was determined by estimating the minimum inhibitory concentration (MIC) and minimal bactericidal/fungicidal concentration (MBC/MFC). To assess the biocompatibility of chitosan/α-Ag 2 WO 4 composites in a range involving MIC and MBC/MFC on keratinocytes cells (NOK-si), an alamarBlue™ assay and an MTT assay were carried out. The SARS-CoV-2 virucidal effects was analyzed in Vero E6 cells through viral titer quantified in cell culture supernatant by PFU/mL assay. Our results showed a very similar antimicrobial activity of chitosan/α-Ag 2 WO 4 3.3 and 6.6, with the last one demonstrating a slightly better action against MSSA. The chitosan/α-Ag 2 WO 4 9.9 showed a wide range of antimicrobial activity (0.49-31.25 µg/mL). The cytotoxicity outcomes by alamarBlue™ revealed that the concentrations of interest (MIC and MBC/MFC) were considered non-cytotoxic to all composites after 72 h of exposure. The Chitosan/α-Ag 2 WO 4 (CS6.6/α-Ag 2 WO 4 ) composite reduced the SARS-CoV-2 viral titer quantification up to 80% of the controls. Then, our results suggest that these composites are highly efficient materials to kill bacteria (Escherichia coli, Methicillin-susceptible Staphylococcus aureus, and the yeast strain Candida albicans), in addition to inactivating SARS-CoV-2 by contact, through ROS production., (© 2022. The Author(s).)

Fungal infections have become a major concern in the medical community, especially those caused by Candida spp. Within this species, Candida albicans stands out for being an opportunistic commensal fungus that can cause superficial and invasive infections. Current antifungal therapy involves the local and/or systemic use of drugs such as azoles, polyenes, and echinocandins. These antifungals are based on highly specific target sites, and the development of resistance may occur with changes in the enzymatic pathways that serve as the drug targets. Thus, the development of new antifungal drugs is highly recommended to prevent drug resistance. The present investigation evaluated the antifungal activity of silver-containing microcrystals such as silver vanadate (α-AgVO 3 ), silver tungstate (α-Ag 2 WO 4 ), and silver molybdate (β-Ag 2 MoO 4 ). In addition to having antimicrobial activity, such compounds should not cause damage to underlying tissues. Thus, to better assess the biocompatibility of new compounds, a new three-dimensional (3D) coculture model involving three cell lines was developed. The validation of the model was based on fluorescent markers and confocal laser microscopy. The biocompatibility of silver-containing microcrystals was evaluated by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. 3D coculture was infected with C. albicans biofilm and challenged with α-AgVO 3 , α-Ag 2 WO 4 , and β-Ag 2 MoO 4 . The action of microcrystals on C. albicans biofilm was evaluated by colony-forming units (CFU/ml) and LIVE/DEAD staining. In addition, production of proinflammatory cytokines interleukin 6 (IL-6), IL-8, IL-1β, and tumor necrosis factor α (TNF-α) was measured by cytometric bead array kit using flow cytometry. The 3D coculture model described here proved to be adequate to assess both the biocompatibility of the new materials and the infectious processes. Regarding the biocompatibility of the microcrystals, only α-AgVO 3 (15.62 µg/ml) showed a decrease in cell viability. The antibiofilm activity of α-Ag 2 WO 4 was similar to that of the standard drug (fluconazole). Although α-Ag 2 WO 4 was able to induce the production of IL-6, IL-8, and IL-1β, no differences in cytokine production were observed between noninfected and infected models treated with this microcrystal. β-Ag 2 MoO 4 inhibits the production of TNF-α in the infected model; however, it showed no antibiofilm activity. Based on the biocompatibility and antifungal findings, α-Ag 2 WO 4 is a promising material for treating C. albicans infection., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Pimentel, Marin-Dett, Assis, Barbugli, Longo and Vergani.)

Polymicrobial biofilms comprising Candida albicans and Staphylococcus aureus can increase the frequency and severity of oral diseases. This study assessed oral keratinocyte cell death, apoptosis and/or necrosis, promoted by soluble factors from single and dual biofilms of S. aureus and C. albicans . The soluble factors were obtained from the 16-h biofilm growth media. Cell viability was assessed by MTT and cell membrane damage by LDH. SEM was used for morphology changes. Assessment of apoptosis and necrosis was performed using annexin V and propidium iodide and caspases -2, -3, -6, -8 and -9. Statistical analysis was conducted with ANOVA and Tukey tests (α = 5%). Dual biofilms promoted the greatest harmful effect on oral cells, with a viability rate of 31.76%, damage to cell membranes and LDH released. Dual biofilms also induced higher percentages of necrotic cells (24.95%). Apoptosis was associated with caspases -2, -3, -6 and -8 activation.

This study demonstrated a drug-delivery system with anionic beta cyclodextrin (β-CD) complexes to retain tetracycline (TC) and control its release from multilayers of poly(acrylic acid) (PAA) and poly(l-lysine) (PLL) in a ten double layers ([PAA/PLL] 10 ) coating onto titanium. The drug-delivery capacity of the multilayer system was proven by controlled drug release over 15 days and sustained released over 30 days. Qualitative images confirmed TC retention within the layer-by-layer (LbL) over 30 days of incubation. Antibacterial activity of TC/anionic β-CD released from the LbL was established against Staphylococcus aureus species. Remarkably, [PAA/PLL] 10 /TC/anionic β-CD antibacterial effect was sustained even after 30 days of incubation. The non-cytotoxic effect of the multilayer system revealed normal human gingival fibroblast growth. It is expected that this novel approach and the chemical concept to improve drug incorporation into the multilayer system will open up possibilities to make the drug release system more applicable to implantable percutaneous devices., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

This study was aimed at analyzing proto-oncogenic signaling pathway activation in normal oral keratinocytes (NOK-si) and neoplastic cell lines (SCC 25 and Detroit 562) stimulated with metabolites (soluble factors) from single and dual biofilms of Candida albicans and Staphylococcus aureus . Soluble factors (SF) from early (16-h) and mature (36-h) biofilms of C. albicans and S. aureus were collected and incubated with cell cultures, which were subsequently evaluated using gene expression via RT-qPCR, cell viability via AlamarBlue TM , and flow cytometry cell cycle analysis. In general, exposure to the SF of early and mature biofilms from C. albicans and dual species caused a major reduction in NOK-si cell viability and enhanced the sub G0 phase. This led to a decrease in gene expression. However, in this cell line, SF of S. aureus biofilms upregulated the CDKN1A gene followed by the maintenance of cell viability and a significant increase in the G2/M population. For tumor cells, SCC 25 and Detroit 562, the stimuli of SF biofilms upregulated oncogenes such as hRAS and mTOR , as well as Bcl-2 and CDKN1A . SCC 25 and Detroit 562 cells could survive even after 24 h of stimuli from both SF (early and mature). This occurred without significant changes taking place in the cell cycle progression for SCC 25, and with a significant tendency to increase the G2/M phase for Detroit 562. These results point to the fact that metabolites from prevalent clinical fungal and bacterial biofilms, C. albicans and S. aureus , can disrupt the homeostasis of normal and neoplastic oral epithelial cells. This changes proto-oncogenes' expression, specifically PI3KCA , hRAS , mTOR , BRAF , and cell cycle genes CDKN1A and Bcl-2 , thus causing a disturbance in cell viability, survival, and the cell cycle profile., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Amaya Arbeláez, de Paula e Silva, Navegante, Valente, Barbugli and Vergani.)

Silver tungstate (Ag 2 WO 4 ) shows structural polymorphism with different crystalline phases, namely, orthorhombic, hexagonal, and cubic structures that are commonly known as α, β, and γ, respectively. In this work, these Ag 2 WO 4 polymorphs were selectively and successfully synthesized through a simple precipitation route at ambient temperature. The polymorph-controlled synthesis was conducted by means of the volumetric ratios of the silver nitrate/tungstate sodium dehydrate precursors in solution. The structural and electronic properties of the as-synthesized Ag 2 WO 4 polymorphs were investigated by using a combination of X-ray diffraction and Rietveld refinements, X-ray absorption spectroscopy, X-ray absorption near-edge structure spectroscopy, field-emission scanning electron microscopy images, and photoluminescence. To complement and rationalize the experimental results, first-principles calculations, at the density functional theory level, were carried out, leading to an unprecedented glimpse into the atomic-level properties of the morphology and the exposed surfaces of Ag 2 WO 4 polymorphs. Following the analysis of the local coordination of Ag and W cations (clusters) at each exposed surface of the three polymorphs, the structure-property relationship between the morphology and the photocatalytic and antibacterial activities against amiloride degradation under ultraviolet light irradiation and methicillin-resistant Staphylococcus aureus , respectively, was investigated. A possible mechanism of the photocatalytic and antibacterial activity as well the formation process and growth of the polymorphs is also explored and proposed.

Purpose: This study investigated the effect of long-term daily chemical disinfection on the topographic and Candida albicans biofilm formation on a denture base resin and a reline acrylic resin., Material and Methods: Circular samples (14 × 1.2 mm) were fabricated from a denture base (Vipi Wave) and reline acrylic resins (Tokuyama Rebase Fast II). Samples were kept in 50 ml of distilled water (48 h at 37°C). Subsequently, the samples were immersed in five different solutions: 0.5% sodium hypochlorite; 3.8% sodium perborate; 2% chlorhexidine gluconate; apple vinegar containing 4% maleic acid; and distilled water (control group). The specimen was immersed in the solutions for 8 h daily and transferred to distilled water at 37°C for more 16 h. The surface topographic and Candida albicans (ATCC 90028) biofilm formation were evaluated at baseline (before chemical disinfection) and after 1, 3 and 6 months of immersion. The surface topographic was evaluated by arithmetical roughness average (Ra) and scanning electron microscope (SEM), while the biofilm formation was evaluated by colony-forming units (CFU/ml) method and Alamar Blue assay (cell metabolism). The results were evaluated by three-way analysis of variance (ANOVAs) and post-hoc tests (α = 0.05)., Results: The results showed statistically significant effects from the type of acrylic resin (p = 0.029) and time (p <0.001) on the roughness of the specimen. In general, the reline resin had higher roughness than the denture base resin. In addition, the roughness of the samples after 1, 3 and 6 months of immersion in the cleaning solutions was higher than at baseline. In relation to the microbiological assays, there were no statistically significant differences (p >0.055) in the CFU/ml values of the biofilms among the different resins, periods of time and cleaning solutions. Considering the metabolism of the cells within the biofilms, the results showed that, at baseline, it was statistically significantly higher (p <0.05) than after 1, 3 and 6 months of storage. The SEM images showed that all disinfectant solutions provided surface changes of both acrylic resins (base and reline) after 1, 3 and 6 months of immersion., Conclusions: The roughness of both acrylic resins was affected by the disinfection in all cleaning agents, increasing over time, and this effect was more evident in the reline acrylic resin group. This surface change was also observed in the SEM images. While the number of cells within the biofilms was not affected by immersion in the cleaning agents, their metabolism was lower after 1, 3 and 6 months of immersion.

Acetohydroxyacid synthase (AHAS, EC 2.2.1.6), the first enzyme in the branched chain amino acid biosynthesis pathway, is the target for more than 50 commercially available herbicides, and is a promising target for antimicrobial drug discovery. Herein, we have expressed and purified AHAS from Candida auris , a newly identified human invasive fungal pathogen. Thirteen AHAS inhibiting herbicides have K i values of <2 μM for this enzyme, with the most potent having K i values of <32 nM. Six of these compounds exhibited MIC 50 values of <1 μM against C. auris (CBS10913 strain) grown in culture, with bensulfuron methyl (BSM) being fungicidal and the most potent (MIC 50 of 0.090 μM) in defined minimal media. The MIC 50 value increases to 0.90 μM in media enriched by the addition of branched-chain amino acids at the expected concentration in the blood serum. The sessile MIC 50 for BSM is 0.6 μM. Thus, it is also an excellent inhibitor of the growth of C. auris biofilms. BSM is nontoxic in HEK-293 cells at concentrations >100 μM and thus possesses a therapeutic index of >100. These data suggest that targeting AHAS is a viable strategy for treating C. auris infections.

Crystal morphology with different surfaces is important for improving the antibacterial activity of materials. In this experimental and theoretical study, the antibacterial activity of β-Ag 2 MoO 4 microcrystals against the Gram-positive bacteria, namely, methicillin-resistant Staphylococcus aureus (MRSA), and the Gram-negative bacteria, namely, Escherichia coli (E. coli), was investigated. In this study, β-Ag 2 MoO 4 crystals with different morphologies were synthetized by a simple co-precipitation method using three different solvents. The antimicrobial efficacy of the obtained microcrystals against both bacteria increased according to the solvent used in the following order: water < ammonia < ethanol. Supported by experimental evidence, a correlation between morphology, surface energy, and antibacterial performance was established. By using the theoretical Wulff construction, which was obtained by means of density functional calculations, the morphologies with large exposition of the (001) surface exhibited superior antibacterial activity. This study provides a low cost route for synthesizing β-Ag 2 MoO 4 crystals and a guideline for enhancing the biological effect of biocides on pathogenic bacteria by the morphological modulation., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The number of studies on microcrystals containing silver has increased in recent decades. Among the silver-containing microcrystals, α-AgVO 3 has gained prominence owing to its polymorphism that allows it to exert interesting antimicrobial activity against pathogenic microorganisms. The aim of this study was to evaluate the antifungal activity and cytotoxicity of three different α-AgVO 3 microcrystals when in solution. α-AgVO 3 microcrystals were synthesized using the co-precipitation method at three different temperatures (10 °C, 20 °C, and 30 °C), and then characterized by X-ray diffraction and scanning electron microscopy. The antifungal activity of α-AgVO 3 microcrystals against Candida albicans was determined by estimating the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC). Fluorescence images were obtained to confirm antifungal concentrations. To assess the biocompatibility of microcrystals applied at MIC and MFC on keratinocytes cells (NOK-si), an Alamar Blue assay, scanning electron microscopy, and a DNA gel integrity test were carried out. The quantitative and qualitative results showed that, regardless of the co-precipitation method used to synthetize α-AgVO 3 microcrystals, C. albicans growth was visibly inhibited at 3.9 μg/mL (MIC) and completely inhibited at 15.62 μg/mL (MFC). The cytotoxic and genotoxic outcomes revealed that the MIC and MFC concentrations did not affect NOK-si cell morphology, proliferation, or DNA integrity. The search for new antimicrobial materials has been the focus of the research community recently because of increases in microbial resistance. The findings reported herein demonstrate a novel antifungal and non-cytotoxic material that could be used in biomedical and dental applications., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The ability to manipulate the structure and function of promising systems via external stimuli is emerging with the development of reconfigurable and programmable multifunctional materials. Increasing antifungal and antitumor activity requires novel, effective treatments to be diligently sought. In this work, the synthesis, characterization, and in vitro biological screening of pure α-Ag 2 WO 4 , irradiated with electrons and with non-focused and focused femtosecond laser beams are reported. We demonstrate, for the first time, that Ag nanoparticles/α-Ag 2 WO 4 composite displays potent antifungal and antitumor activity. This composite had an extreme low inhibition concentration against Candida albicans, cause the modulation of α-Ag 2 WO 4 perform the fungicidal activity more efficient. For tumor activity, it was found that the composite showed a high selectivity against the cancer cells (MB49), thus depleting the populations of cancer cells by necrosis and apoptosis, without the healthy cells (BALB/3T3) being affected.

Background: Evaluate the effect of low-temperature plasma (LTP) on an anaerobic biofilm and on the biological response of an in vitro reconstituted gingival epithelium tissue., Methods: Porphyromonas gingivalis W83 biofilm was cultured on titanium discs and reconstituted gingival tissues were submitted to similar treatment conditions., Treatments: LTP1-plasma treatment for 1 minute, LTP3-plasma treatment for 3 minute, CHX-0.2% chlorhexidine for 1 minute, GAS-gas only (no plasma) for 3 minute, and NEGATIVE-no treatment. TRITON group was included as a positive control for tissue analysis. Counting of viable colony forming units (CFU/mL) and confocal laser scanning microscopy were performed to evaluate LTP's antimicrobial effect. EpiGingival tissue was evaluated through cytotoxocity, viability, histology, and imunnohistochemistry (Ki67, vascular endothelial growth factor-A vascular endothelial growth factor A [VEGF-A], and terminal deoxynucleotidyl transferase dUTP nick end labeling terminal deoxynucleotidyl transferase dutp nick end labeling [TUNEL] expression)., Results: LTP1 and LTP3 presented significantly different reduced CFU/mL reduction in comparison to the negative control (Ρ < 0.001), but it was not as effective as the positive control (CHX). Low cytotoxicity and high viability were observed in gingival epithelium of NEGATIVE, GAS, CHX, and both LTP groups. The morphologic analysis of gingival epithelium revealed minor cell damage in the plasma groups (score 1). LTP1, LTP3, GAS, and NEGATIVE groups exhibited less than 5% of basal layer positive cells. LTP1, LTP3, GAS, and CHX groups were not positive for TUNEL assay. LTP1 and LTP3 showed the most positivity for VEGF., Conclusions: LTP treatment can be considered as an effective method for reducing P. gingivalis biofilm on implant surfaces, while being safe for the gingival epithelium. Furthermore, plasma treatment may be associated with cell repair., (© 2018 American Academy of Periodontology.)

Silver tungstate (α-Ag 2 WO 4 ) microcrystals have shown encouraging results regarding their antimicrobial activity. However, in addition to the promising outcomes in fighting oral disease, cytotoxic tests are mandatory for screening new materials for biological applications. Here, we developed a better understanding of the effects of microcrystals on the behavior of both human gingival fibroblast (HGF) cells and three-dimensional (3D) collagen matrices. To perform these experiments, the lowest concentration of α-Ag 2 WO 4 capable of preventing the visible growth of Candida albicans (C. albicans) planktonic cells was defined as the test concentration, and it ranged from 0.781 (C1) to 7.81 (C2) to 78.1 (C3) μg/mL. Complete medium and lysis buffer (LB) served as negative (C - ) and positive (C + ) controls, respectively. The effect of the microcrystal concentration on the morphology, remodeling and proliferation of HGF cells was evaluated by different approaches. Quantitative and qualitative assessments demonstrated that α-Ag 2 WO 4 did not affect the mitochondrial enzymatic activity of HGF cells cultured in a monolayer or the cell viability within 3D collagen matrices. These experiments showed that α-Ag 2 WO 4 at the C2 concentration did not damage the genomic DNA. The development of new materials is attractive for the possible treatment of diseases and for avoiding indiscriminate prescribing of antibiotics. These findings provide information on the effect of α-Ag 2 WO 4 on cell behavior and reveal that these microcrystals are non-cytotoxic against human gingival cells over a sufficient period to measure the hazard potential., (Copyright © 2018. Published by Elsevier B.V.)

The aim of this study was to (i) design, develop and validate a practical and physiologically relevant reconstituted in vitro oral mucosa tissue model and (ii) to assess its applicability in in vitro host-pathogen interactions with C. albicans and S. aureus. Co-culture organotypic constructions were created by incorporating specific numbers of keratinocytes (NOK-si) onto cellularised, collagen gel scaffolds containing human gingival fibroblasts incubated in KGM media and cultured for 14 days. The detection of the appropriate oral mucosa/epithelial structure was evaluated by histology (hematoxylin and eosin (HE), periodic acid-Schiff (P.A.S.) and Picrosirius red), and immunocytochemistry (cytokeratin 13, cytokeratin 14, Ki-67 and collagen IV) compared to a normal human gingiva. The morphology of the reconstituted tissue was analyzed by Transmission Electron Microscopy. To further quantitate tissue damage, lactate dehydrogenase (LDH) was measured in the tissue supernatant. NOK-si grown upon a gingival scaffold provided an organotypic model in an in vitro setting and exhibited structural characteristics typically associated with normal oral mucosa. Immunocytochemistry revealed the detection of epithelial cytokeratins 13 and 14, Col IV and Ki-67 in the reconstituted oral mucosa model. Infection was detected after 8 h and 16 h. This study presents an in vitro cellularised, organotypic model of reconstituted oral mucosa, which enables close control and characterization of its structure and differentiation over a mid-length period of time in culture., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Bimetallic nanoalloys with a wide variety of structures and compositions have been fabricated through many diverse techniques. Generally, various steps and chemicals are involved in their fabrication. In this study, the synthesis of Ag-Bi nanoalloys by femtosecond laser irradiation of an inorganic oxide Ag 2 WO 4 /NaBiO 3 target without any chemicals like reducing agents or solvent is presented. The interaction between these materials and the ultrashort pulse of light allows the migration of Ag and Bi atoms from the crystal lattice to the particles surfaces and then to the plasma plume, where the reduction of the positively charged Ag and Bi species in their respective metallic species takes place. Subsequently, the controlled nucleation and growth of the Ag-Bi alloyed nanoparticles occurs in situ during the irradiation process in air. Although at the bulk level, these elements are highly immiscible, it was experimentally demonstrated that at nanoscale, the Ag-Bi nanoalloy can assume a randomly mixed structure with up to 6 ± 1 atom % of Bi solubilized into the face-centered cubic structure of Ag. Furthermore, the Ag-Bi binary system possesses high antibacterial activity against Staphylococcus aureus (methicillin-resistant and methicilin-susceptible), which is interesting for potential antimicrobial applications, consequently increasing their range of applicability. The present results provide potential insights into the structures formed by the Ag-Bi systems at the nanoscale and reveal a new processing method where complex inorganic oxides can be used as precursors for the controlled synthesis of alloyed bimetallic nanoparticles., Competing Interests: The authors declare no competing financial interest.

Objectives: The morphology and chemical composition of enamel submitted to different sterilization methods was studied., Methods: X-ray photoelectron spectroscopy (XPS), field emission gun scanning electron microscopy (FEG-SEM), and energy-dispersive X-ray spectroscopy (EDS) were performed to evaluate 50 bovine enamel specimens sterilized using four methods: microwaving (MI), gamma irradiation (GI), ethylene oxide (EO), and steam autoclave (SA). Non-sterilized specimens were used as control., Results: XPS indicated that the concentration of P (phosphorus), CO 3 (carbonate), and CO 3 /P was not changed in all groups. GI produced no significant change on elemental composition. SA produced the major decrease in calcium (Ca), Ca/P ratio, and increase in N (nitrogen). MI was found to decrease Ca, Ca/P ratio and O (oxygen), and increase in C (carbon) and N. EO produced decrease in Ca and O with increased C concentration. FEG-SEM revealed surface and in-depth morphological changes on SA specimens. Minor surface alterations were observed for EO and for MI groups, and no alteration was observed on GI group. EDS indicated no difference on elemental composition of enamel bulk among groups., Conclusions: SA produced mineral loss and morphological alterations on surface and in depth. MI and EO sterilization caused mineral loss showing only slight alteration on enamel surface. GI sterilization preserves the morphological characteristics of enamel. The sterilization methods could be classified from lower to high damage as GI < MI < EO < SA., Clinical Relevance: This is a comprehensive comparative study where different methods for enamel sterilization were investigated in terms of chemical changes. The results presented here may help researchers to choose the most appropriate method for their research setting and purpose.

In recent years, complex nanocomposites formed by Ag nanoparticles coupled to an α-Ag 2 WO 4 semiconductor network have emerged as promising bactericides, where the semiconductor attracts bacterial agents and Ag nanoparticles neutralize them. However, the production rate of such materials has been limited to transmission electron microscope processing, making it difficult to cross the barrier from basic research to real applications. The interaction between pulsed laser radiation and α-Ag 2 WO 4 has revealed a new processing alternative to scale up the production of the nanocomposite resulting in a 32-fold improvement of bactericidal performance, and at the same time obtaining a new class of spherical Ag x W y O z nanoparticles.

Statement of Problem: Peri-implantitis is considered the most important biological complication responsible for late implant failure. The physical chemical properties intrinsic to each material can affect the first step to biofilm development and is an important precursor to the adaptive behavior of pathogenic bacteria species., Purpose: The purpose of this in vitro study was to evaluate the effect of 2 commercially available implant abutment materials on the adhesion phase and biofilm formation., Material and Methods: Disks (8 mm in diameter, 2 mm thick) of machined pure titanium (Ti) and yttrium-stabilized zirconia (ZrO 2 ) materials were used to mimic implant abutments. The physical chemical surface properties were investigated using different approaches. Initial adherent bacteria and biofilm formation were evaluated after 16 and 48 hours by incubating the disks in a rich medium containing representative saliva-derived oral microbial community. Unpaired t test, 2 tailed, was used to compare the groups., Results: Ti presented lower hydrophobicity and surface free energy values than the ZrO 2 , and 6.1-fold fewer bacteria adhered to the Ti. After 48 hours, detailed quantitative analysis showed that biofilm biomass and biofilm density were lower on the Ti disks than on ZrO 2 . The quantity of phylotypes on the Ti and ZrO 2 surfaces was relatively similar during the attachment and early biofilm formation periods., Conclusions: Although no difference in the bacteria profile was observed between both materials independent of the time point, the highest level of colonization was on ZrO 2 ., (Copyright © 2017 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.)

Purpose: This clinical study evaluated the effect of microwave disinfection protocols on the occlusal pressure pattern of dentures., Materials and Methods: Dentures were constructed for 40 patients and evaluated as follows (n = 20). Group 1: Patients had the maxillary dentures submitted to microwave disinfection, once a week, for 4 weeks. Group 2: Patients had the maxillary dentures submitted to microwave disinfection, three times a week, for 4 weeks. Occlusal contacts were recorded on five occasions: 30 days after denture insertion and before first disinfection (baseline or control group); 1 week after disinfection; 2 weeks after disinfection; 3 weeks after disinfection; 4 weeks after disinfection. Occlusal contacts were analyzed by T-Scan III. Intergroup analysis was performed using the Mann-Whitney test and intragroup analysis using the Friedman test with significance of 5%., Results: The results showed no significant difference between groups during the periods. The data on parameters loss of denture adaptation or complaints showed that patients used their dentures regularly for eating and expressed comfort and satisfaction in all experimental periods. The evaluation of functional occlusion revealed that the distribution of the occlusal contacts remained unaltered after disinfection., Conclusion: Microwave disinfection protocols as studied in this report did not influence occlusal contacts of the complete dentures., (© 2015 by the American College of Prosthodontists.)

Background: The objective of this study was to better understand the effects of soluble factors from biofilm of single- and mixed-species Candida albicans (C. albicans) and methicillin-sensitive Staphylococcus aureus (MSSA) cultures after 36 h in culture on keratinocytes (NOK-si and HaCaT) and macrophages (J774A.1). Soluble factors from biofilms of C. albicans and MSSA were collected and incubated with keratinocytes and macrophages, which were subsequently evaluated by cell viability assays (MTT). Lactate dehydrogenase (LDH) enzyme release was measured to assess cell membrane damage to keratinocytes. Cells were analysed by brightfield microscopy after 2 and 24 h of exposure to the soluble factors from biofilm. Cell death was detected by labelling apoptotic cells with annexin V and necrotic cells with propidium iodide (PI) and was visualized via fluorescence microscopy. Soluble factors from biofilm were incubated with J774A.1 cells for 24 h; the subsequent production of NO and the cytokines IL-6 and TNF-α was measured by ELISA., Results: The cell viability assays showed that the soluble factors of single-species C. albicans cultures were as toxic as the soluble factors from biofilm of mixed cultures, whereas the soluble factors of MSSA cultures were less toxic than those of C. albicans or mixed cultures. The soluble factors from biofilm of mixed cultures were the most toxic to the NOK-si and HaCaT cells, as confirmed by analyses of PI labelling and cell morphology. Soluble factors from biofilm of single-species MSSA and mixed-species cultures induced the production of IL-6, NO and TNF-α by J744A.1 macrophages. The production of IL-6 and NO induced by the soluble factors from biofilm of mixed cultures was lower than that induced by the soluble factors from biofilm of single-species MSSA cultures, whereas the soluble factors from biofilm of C. albicans cultures induced only low levels of NO., Conclusions: Soluble factors from 36-h-old biofilm of C. albicans and MSSA cultures promoted cell death and inflammatory responses.

The electronic configuration, morphology, optical features, and antibacterial activity of metastable α-AgVO 3 crystals have been discussed by a conciliation and association of the results acquired by experimental procedures and first-principles calculations. The α-AgVO 3 powders were synthesized using a coprecipitation method at 10, 20, and 30 °C. By using a Wulff construction for all relevant low-index surfaces [(100), (010), (001), (110), (011), (101), and (111)], the fine-tuning of the desired morphologies can be achieved by controlling the values of the surface energies, thereby lending a microscopic understanding to the experimental results. The as-synthesized α-AgVO 3 crystals display a high antibacterial activity against methicillin-resistant Staphylococcus aureus. The results obtained from the experimental and theoretical techniques allow us to propose a mechanism for understanding the relationship between the morphological changes and antimicrobial performance of α-AgVO 3 .

According to the American Academy of Implant Dentistry, 3 million Americans have dental implants, and this number is growing by 500,000 each year. Proportionally, the number of biological complications is also increasing. Among them, peri-implant disease is considered the most common cause of implant loss after osseointegration. In this context, microorganisms residing on the surfaces of implants and their prosthetic components are considered to be the primary etiologic factor for peri-implantitis. Some research groups have proposed combining surgical and non-surgical therapies with systemic antibiotics. The major problem associated with the use of antibiotics to treat peri-implantitis is that microorganisms replicate very quickly. Moreover, inappropriate prescription of antibiotics is not only associated with potential resistance but also and most importantly with the development of superinfections that are difficult to eradicate. Although antimicrobial photodynamic therapy (aPDT) was discovered several years ago, aPDT has only recently emerged as a possible alternative therapy against different oral pathogens causing peri-implantitis. The mechanism of action of aPDT is based on a combination of a photosensitizer drug and light of a specific wavelength in the presence of oxygen. The reaction between light and oxygen produces toxic forms of oxygen species that can kill microbial cells. This mechanism is crucial to the efficacy of aPDT. To help us understand conflicting data, it is necessary to know all the particularities of the etiology of peri-implantitis and the aPDT compounds. We believe that this review will draw attention to new insights regarding the impact of aPDT on peri-implant disease., (Copyright © 2016 Elsevier B.V. All rights reserved.)

This study evaluated the potential of curcumin-mediated antimicrobial photodynamic inactivation (API) on multispecies biofilms of Candida albicans, Candida glabrata, and Streptococcus mutans of different ages. Acrylic samples (n = 480) were made with standardized rough surfaces and incubated with bacteria and yeast for 24 or 48 h. API was performed with curcumin (80, 100, 120 μM) and LED light. Additional acrylic samples were treated with curcumin or LED light only. Positive control samples received neither light nor curcumin. After API, colony counts were quantified (CFU/mL), cell metabolism was determined by means of XTT assay, and the total biofilm biomass was evaluated using Crystal Violet (CV) staining assay and images were obtained by confocal laser scanning microscopy (CLSM). The data were analyzed by nonparametric two-way ANOVA and post hoc Tukey tests (α < 0.05). For 24-h biofilm, API resulted in statistically significant difference (ρ < 0.001) of viability of C. albicans compared with control (P-L-) for all Cur concentrations. For 48-h biofilm, API resulted in statistically significant difference (ρ < 0.001) compared with control only when Cur at 120 μM was used. API promoted statistically significant difference (ρ ≤ 0.001) in the viability of S. mutans and C. glabrata for all Cur concentrations in the two biofilm ages. In addition, API produced a statistically significant difference (ρ < 0.001) of metabolic activity and of total biomass (ρ < 0.001) of multispecies biofilms compared with control for all Cur concentrations. It can be concluded that both 24- and 48-h biofilms were susceptible to API mediated by Cur; however, 24-h biofilm was more sensitive than the 48-h biofilm.

In this study, we investigate the structure, antifungal activity, and optical properties of β-Ag2MoO4 using experimental and theoretical approaches. β-Ag2MoO4 samples were prepared by a co-precipitation method using different solvents (water, ethanol and ammonia), and their antifungal activity against Candida albicans was investigated. The samples were characterized by X-ray diffraction, micro-Raman spectroscopy, field emission scanning electron microscopy, and transmission electron microscopy with energy dispersive spectroscopy. The optical properties were investigated by UV-Vis spectroscopy and photoluminescence measurements at room temperature. The thermodynamic equilibrium shape of the β-Ag2MoO4 crystals was determined based on the surface energies calculated using Wulff construction. The (011) orientation was the predominant surface in the morphology. The experimental morphology was obtained by varying the surface energy ratio for each facet. A large decrease in surface energy for the (111) surface provided the experimental morphology for crystals synthesized using water and ethanol as solvents; when the surface energies for both (011) and (001) surfaces increased, the crystal morphology obtained using ammonia as a solvent was reproduced. A correlation between the exposed surfaces and antifungal activity was revealed, and an explanation to this behavior that arises from different morphologies and structural data was provided. Theoretical calculations confirm the rationality of the experimental scheme and elucidate the underlying reason for the fungistatic and fungicidal activity against Candida albicans.

This study assessed the effect of the buffers 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) and 3-(N-morpholino) propanesulfonic acid (MOPS) on keratinocyte cell viability and microbial growth. It was observed that RPMI buffered with HEPES, supplemented with l-glutamine and sodium bicarbonate, can be used as a more suitable medium to promote co-culture., (Copyright © 2016. Published by Elsevier B.V.)

Considering the ability of atmospheric-pressure cold plasma (ACP) to disrupt the biofilm matrix and rupture cell structure, it can be an efficient tool against virulent oral biofilms. However, it is fundamental that ACP does not cause damage to oral tissue. So, this study evaluated (1) the antimicrobial effect of ACP on single- and dual-species biofilms of Candida albicans and Staphylococcus aureus as well as (2) the biological safety of ACP on in vitro reconstituted oral epithelium. Standardized cell suspensions of each microorganism were prepared for biofilm culture on acrylic resin discs at 37°C for 48 hours. The biofilms were submitted to ACP treatment at 10 mm of plasma tip-to-sample distance during 60 seconds. Positive controls were penicillin G and fluconazole for S. aureus and C. albicans, respectively. The biofilms were analyzed through counting of viable colonies, confocal laser scanning microscopy, scanning electron microscopy and fluorescence microscopy for detection of reactive oxygen species. The in vitro reconstituted oral epithelium was submitted to similar ACP treatment and analyzed through histology, cytotoxocity test (LDH release), viability test (MTT assay) and imunnohistochemistry (Ki67 expression). All plasma-treated biofilms presented significant log10 CFU/mL reduction, alteration in microorganism/biofilm morphology, and reduced viability in comparison to negative and positive controls. In addition, fluorescence microscopy revealed presence of reactive oxygen species in all plasma-treated biofilms. Low cytotoxicity and high viability were observed in oral epithelium of negative control and plasma group. Histology showed neither sign of necrosis nor significant alteration in plasma-treated epithelium. Ki67-positive cells revealed maintenance of cell proliferation in plasma-treated epithelium. Atmospheric-pressure cold plasma is a promissing approach to eliminate single- and dual-species biofilms of C. albicans and S. aureus without having toxic effects in oral epithelium.

This study assessed the cytotoxicity of antimicrobial Photodynamic Inactivation (aPDI), mediated by curcumin, using human keratinocytes co-cultured with Candida albicans. Cells and microorganisms were grown separately for 24 hours and then kept in contact for an additional 24 hours. After this period, aPDI was applied. The conditions tested were: P+L+ (experimental group aPDI); P-L+ (light emitting diode [LED] group); P+L- (curcumin group); and P-L- (cells in co-culture without curcumin nor LED). In addition, keratinocytes and C. albicans were grown separately, were not placed in the co-culture and did not receive aPDI (control group). Cell proliferation was assessed using Alamar Blue, MTT, XTT and CFU tests. Qualitative and quantitative analyses were performed. Analysis of variance (ANOVA) was applied to the survival percentages of cells compared to the control group (considered as 100% viability), complemented by multiple comparisons using Tukey's test. A 5% significance level was adopted. The results of this study showed no interference in the metabolism of the cells in co-culture, since no differences were observed between the control group (cultured cells by themselves) and the P-L- group (co-culture cells without aPDI). The aPDI group reached the highest reduction (p = 0.009), which was equivalent to 1.7 log10 when compared to the control group. The P+L-, P-L+, P-L- and control groups were not statistically different (ρ > 0.05). aPDI inhibited the growth of keratinocytes and C. albicans in all tests, so the therapy was considered slightly (inhibition between 25 and 50% compared to the control group) to moderately (inhibition between 50 and 75% compared to the control group) cytotoxic.

Aim: The aim of the present study was to evaluate the effect of surface roughness (roughness average [Ra] μm) on the hydrophobicity of a denture-base acrylic resin and the initial adherence and biofilm formation of Candida albicans (C. albicans)., Methods: Disk-shaped specimens were divided into six groups: Ra 0.05, Ra 0.2, Ra 0.4, Ra 0.8, Ra 1.5, and Ra 3.0. Water contact angles (WCA) were measured, and the specimens incubated with C. albicans for 90 min (initial adherence, n = 108) or 48 h (biofilm formation, n = 108). Adhered and biofilm cells were evaluated by c.f.u./mL and 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT), and the correlation between the two methods was evaluated. The surface of the specimens and cells (adhered and biofilm) were also analyzed by scanning electron microscopy (SEM)., Results: Groups Ra 0.05 and 3.0 exhibited the lowest (~75°) and the highest (~100°) WCA mean values, respectively. For both initial adherence and biofilm formation, no statistically-significant differences were observed among all groups, as determined by c.f.u./mL and XTT. A positive correlation between these two methods was found. SEM analysis showed the presence of scratches and valleys on the acrylic specimens and densely-packed yeast cells covering the entire surface., Conclusions: Roughness significantly increased hydrophobicity (WCA), but had no effect on the number and metabolic activity of adherent and biofilm cells of C. albicans., (© 2014 Wiley Publishing Asia Pty Ltd.)

Statement of Problem: The longevity of dental implants depends on the absence of inflammation in the periimplant tissue. Similar to teeth, pathogenic bacteria can adhere on implant abutment surfaces and cause periimplant disease and consequently implant loss., Purpose: The purpose of this in vitro study was to evaluate the influence of physical and chemical properties of 2 common materials used as implant abutments, titanium (Ti) and zirconia (ZrO2), and the use of bovine enamel (BE) as a positive control on biofilm formation., Material and Methods: Biofilm formation was analyzed by growing Porphyromonas gingivalis and Fusobacterium nucleatum as monospecies and mixed species biofilms on the surfaces. The mean roughness (Ra) and surface free energy were evaluated for each material. Mature biofilm, formed after 7 days of incubation, was analyzed quantitatively and qualitatively by colony-forming unit and confocal laser scanning microscopy., Results: The mean roughness in all disks was ≤0.21 μm and did not affect the bacterial adhesion. Titanium showed a greater degree of hydrophilicity compared with BE after 90 minutes of immersion in saliva. The surface free energy did not show differences, with the highest values for BE. Monospecies biofilms formed by P. gingivalis on Ti, and mixed species biofilm on ZrO2 exhibited small numbers of cells on disk surfaces. By confocal imaging, the mixed species biofilm appeared as a thin layer on ZrO2 surfaces., Conclusions: Material surfaces could have a significant impact on biofilm formation. ZrO2 implant abutment surfaces showed a decrease in anaerobic biofilm compared with Ti and BE., (Copyright © 2016 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.)

Surface attachment is the first step in biofilm formation, and the ability of bacteria to adhere to surfaces and develop a biofilm is directly influenced by electrostatic interactions between the bacteria and the chemical composition of material surfaces. Here, we investigated the influence of physical and chemical characteristics of titanium (Ti) and zirconia (ZrO2) as implant abutment surfaces on the bacterial adhesion phase and compared the results to bovine enamel (BE) simulating a human tooth. To achieve this goal, we used 2 common pathogens of the oral cavity, Streptococcus mutans UA140 and Porphyromonas gingivalis 33277. To investigate the influence of material surfaces on bacterial adhesion, we studied the surface free energy as well as the topography by atomic force microscopy, and the chemical elements composition by scanning electron microscopy equipped with an energy dispersive X-ray spectroscope. Our results indicated a hydrophobic characteristic for all of the materials; however, the presence of polar and nonpolar components could aid in understanding why greater numbers of bacteria had adhered to BE compared to the other surfaces. Our confocal microscopy data support the proposition that electrostatic interactions, indeed, affected the initial adhesion phase. Within the limitations of a laboratory study, the results revealed bacterial adhered on BE and no bacteria could be observed by confocal images on Ti and ZrO2 implant abutment surfaces.

The aim of this study was to establish a reproducible protocol using the methodology of hyaline zones around the colonies on specific agar plates for phospholipase and proteinase production. This was an in vitro double-blind experiment, in which the dependent variables were the enzymatic activity measurements (Pz) for the production of phospholipase (Pz-ph) and the production of secreted aspartyl proteinases (Pz-sap). Three independent variables give rise to different measurement protocols. All measurements were carried out at two different moments by four examiners (E1, E2, E3, and E4). The minimum sample size was 30 Candida albicans clinical isolates. Specific agar plates for phospholipase and SAPs production were prepared according the literature. The intra-and inter-examiner reproducibility for each protocol was estimated using the Intraclass Correlation Coefficient (ICC) and its confidence interval (95% CI). Based on the results obtained for both phospholipase and SAPs, there appears to be no consensus on the protocol chosen for each particular examiner. Measuring the colonies in triplicate may be the main factor associated with the increase in measurement accuracy and should therefore take precedence over measuring only one colony. When only one examiner is responsible for taking measurements, a standard protocol should be put in place and the statistical calibration of this researcher should be done prior to data collection. However, if two or more researchers are involved in the assessment of agar plates, our results suggest that the protocols using software to undertake plate reading is preferred., (© The Author 2015. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Polymicrobial biofilms are an understudied and a clinically relevant problem. This study evaluates the interaction between C. albicans, and methicillin- susceptible (MSSA) and resistant (MRSA) S. aureus growing in single- and dual-species biofilms. Single and dual species adhesion (90 min) and biofilms (12, 24, and 48 h) were evaluated by complementary methods: counting colony-forming units (CFU mL-1), XTT-reduction, and crystal violet staining (CV). The secretion of hydrolytic enzymes by the 48 h biofilms was also evaluated using fluorimetric kits. Scanning electron microscopy (SEM) was used to assess biofilm structure. The results from quantification assays were compared using two-way ANOVAs with Tukey post-hoc tests, while data from enzymatic activities were analyzed by one-way Welch-ANOVA followed by Games-Howell post hoc test (α = 0.05). C. albicans, MSSA and MRSA were able to adhere and to form biofilm in both single or mixed cultures. In general, all microorganisms in both growth conditions showed a gradual increase in the number of cells and metabolic activity over time, reaching peak values between 12 h and 48 h (ρ<0.05). C. albicans single- and dual-biofilms had significantly higher total biomass values (ρ<0.05) than single biofilms of bacteria. Except for single MRSA biofilms, all microorganisms in both growth conditions secreted proteinase and phospholipase-C. SEM images revealed extensive adherence of bacteria to hyphal elements of C. albicans. C. albicans, MSSA, and MRSA can co-exist in biofilms without antagonism and in an apparent synergistic effect, with bacteria cells preferentially associated to C. albicans hyphal forms.

Antimicrobial photodynamic therapy represents an alternative method of killing resistant pathogens. Efforts have been made to develop delivery systems for hydrophobic drugs to improve the photokilling. This study evaluated the photodynamic effect of chloro-aluminum phthalocyanine (ClAlPc) encapsulated in nanoemulsions (NE) on methicillin-susceptible and methicillin-resistant Staphylococcus aureus suspensions and biofilms. Suspensions and biofilms were treated with different delivery systems containing ClAlPc. After the pre-incubation period, the drug was washed-out and irradiation was performed with LED source (660 ± 3 nm). Negative control samples were not exposed to ClAlPc or light. For the suspensions, colonies were counted (colony-forming units per milliliter (CFU/mL)). The metabolic activity of S. aureus suspensions and biofilms were evaluated by the XTT assay. The efficiency was dependent on the delivery system, superficial load and light dose. Cationic NE-ClAlPc and free-ClAlPc caused photokilling of the both strains of S. aureus. For biofilms, cationic NE-ClAlPc reduced cell metabolism by 80 and 73% of susceptible and resistant strains, respectively. Although anionic NE-ClAlPc caused a significant CFU/ml reduction for MSSA and MRSA, it was not capable of reducing MRSA biofilm metabolism. This therapy may represent an alternative treatment for eradicating resistant strains.

This study is a framework proposal for understanding the antimicrobacterial effect of both α-Ag2WO4 microcrystals (AWO) synthesized using a microwave hydrothermal (MH) method and α-Ag2WO4 microcrystals with Ag metallic nanofilaments (AWO:Ag) obtained by irradiation employing an electron beam to combat against planktonic cells of methicillin-resistant Staphylococcus aureus (MRSA). These samples were characterized by X-ray diffraction (XRD), FT-Raman spectroscopy, ultraviolet visible (UV-vis) measurements, field emission scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM), and high resolution transmission electron microscopy (HRTEM). The results reveal that both AWO and AWO:Ag solutions have bacteriostatic and bactericidal effects, but the irradiated sample is more efficient; i.e., a 4-fold of the MRSA planktonic cells as compared to the nonirradiated sample was observed. In addition, first principles calculations were performed to obtain structural and electronic properties of AWO and metallic Ag, which provides strong quantitative support for an antimicrobacterial mechanism based on the enhancement of electron transfer processes between α-Ag2WO4 and Ag nanoparticles.

Objective: To evaluate the expression of phospholipase (PL) and secreted aspartyl proteinase (SAP) by Candida glabrata and C tropicalis obtained from the denture biofilms of healthy participants (16 isolates), patients with oral candidiasis with diabetes (10 isolates), and patients with oral candidiasis without diabetes (25 isolates)., Study Design: After incubation, the supernatants and pellets of the isolates were used for the enzymatic assays and quantification of colony-forming units (CFU), respectively. Colorimetric tests were used with phosphatidylcholine as a substrate for PL and azocasein as a substrate for SAP, and the absorbances of the samples were measured. Enzymatic rates were calculated, and values were normalized by CFU. Results were analyzed with factorial analyses of variance (α = .05)., Results: C tropicalis and C glabrata were proteolytic and phospholipolytic. The clinical sources of isolates had no significant effect on the enzymatic activities (P > .05). C tropicalis had significantly higher enzymatic activity for both PL and SAP (P < .001) than did C glabrata., Conclusions: C tropicalis isolates produced significantly higher amounts of both enzymes than did the C glabrata isolates., (Copyright © 2014 Elsevier Inc. All rights reserved.)