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Objective:: To determine the occurrence and the in vitro antimicrobial susceptibility of enteric rods and pseudomonads from the denture biofilm of 52 subjects at the Center for Dental Specialties of Sobral/ Ceara, Brazil., Material and Methods:: Denture biofilm was collected and samples plated on MacConkey agar. The isolated bacterial colonies were identified using the BBL Crystal enteric/non-fermenter system. Antibiotic bacterial susceptibility was assessed by the disc diffusion method of amoxicillin, amoxicillin/clavulanic acid, doxycycline, tetracycline, tobramycin, imipenem, cefotaxime, and ciprofloxacin. The Minimum Inhibitory Concentration (MIC) of cefotaxime, tobramycin, doxycycline, imipenem, and ciprofloxacin was determined for 40 species by E-test., Results:: 34 subjects (65.4%) harbored enteric rods in their prostheses. Klebsiella pneumoniae (26.5%), Escherichia coli (23.5%), and Enterobacter aerogenes (23.5%) were the most prevalent species. All organisms were susceptible to ciprofloxacin and most species were resistant to amoxicillin or amoxicillin/clavulanic acid, demonstrating variable sensitivity patterns to other antimicrobials. However, the MIC showed the emergence of strains with reduced sensitivity to ciprofloxacin (MIC90≥3 μg/ mL) and cefotaxime (MIC90≥2 μg/mL)., Conclusion:: The findings show high prevalence of nosocomial diseases-related bacterial species and low susceptibility to antimicrobial drugs. Therefore, these results imply caution against the indiscriminate use of broad spectrum antibiotics in dental practice.

Background: Some authors have reported the antimicrobial action of photodynamic antimicrobial chemotherapy (PACT) on bacteria related to nosocomial infections but there are few studies evaluating PACT on Serratia marcescens grown as planktonic cultures or as biofilms. The purpose of this study was to analyze the S. marcescens resistance profile and its susceptibility to PACT., Methods: Initially, 55 S. marcescens strains isolated from environmental, oral and extra-oral infections were tested by antimicrobial resistance to cefotaxime (CTX), imipenem (IPM), ciprofloxacin (CIP), tobramycin (TOB) and doxycycline (DOX) using E-test(®). Following, isolates grown as planktonic cultures or biofilms were submitted to PACT using the association of a light-emitting diode and toluidine blue (TBO). The E-test(®) results demonstrated intermediated sensitive strains to CTX, IMP, TOB, and DOX; and resistant strains to CTX, TOB, DOX and CIP. Also, CTX and IMP demonstrated variation when CLSI 2007 and CLSI 2015 were compared., Results: Planktonic cultures and biofilms submitted to PACT demonstrated counts varying from 10(11) to 10(7) for planktonic cultures and 10(10) to 10(7) for biofilms. There were no statistical differences in the results when planktonic cultures and biofilms were compared., Conclusions: Increase in the profile of S. marcescens resistance was observed when CLSI 2007 and CLSI 2015 were compared. Also, IMP remains as the drug with lower rate of resistance. Additionally, both S. marcescens planktonic cultures and early biofilms are susceptible to PACT under tested conditions., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Background: Ultraconservative removal of carious tissue is becoming increasingly highlighted for management of deep caries lesions, and combined with an antimicrobial photochemistry-based treatment modality (PACT), this approach can be enhanced favoring dental tissue repair and preservation. The aim was to investigate the effectiveness of PACT using a light emitting diode (LED) associated with a photosensitizer toluidine blue ortho (TBO) on deep caries lesions., Methods: For that, a single blind, randomized, controlled, split-mouth clinical trial where 45 patients with at least two deep carious lesions on permanent posterior teeth was performed. The primary intervention was deep caries lesion management with disinfection of remaining dentin tissue using PACT. Bacterial counts were measured following treatments as the main outcome. The remaining dentinal samples of each lesion were treated with either non-PACT-control or PACT. The PACT procedure were characterized by 100 μg mL(-1) TBO followed by 94J cm(-2) LED irradiation. Samples of dentin were collected before and immediately after treatments for microbiological analysis of total viable bacteria, mutans streptococci and Lactobacillus spp. counts. Microbial reduction was data were submitted to unpaired t test (α=5%)., Results: PACT led to statistically significant reductions in mutans streptococci (1.08 ± 1.20 log), Lactobacillus spp. (1.69 ± 1.37 log), and total viable bacteria (1.07 ± 1.01 log) compared to the control, which showed log reductions respectively of 0.05 ± 0.49, 0.52 ± 0.89, and 0.47 ± 0.77 for the same microorganisms., Conclusion: Dentin from deep carious lesions treated with PACT showed a decrease in cariogenic microbial load., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Background: The use of blue light has been proposed as a direct means of affecting local bacterial infections, however the use of blue light without a photosensitizer to prevent the biofilm development has not yet been explored. The aim of this study was to determine how the twice-daily treatment with blue light affects the development and composition of a matrix-rich biofilm., Methodology/principal Findings: Biofilms of Streptococcus mutans UA159 were formed on saliva-coated hydroxyapatite discs for 5 days. The biofilms were exposed twice-daily to non-coherent blue light (LumaCare; 420 nm) without a photosensitizer. The distance between the light and the sample was 1.0 cm; energy density of 72 J cm-2; and exposure time of 12 min 56 s. Positive and negative controls were twice-daily 0.12% chlorhexidine (CHX) and 0.89% NaCl, respectively. Biofilms were analyzed for bacterial viability, dry-weight, and extra (EPS-insoluble and soluble) and intracellular (IPS) polysaccharides. Variable pressure scanning electron microscopy and confocal scanning laser microscopy were used to check biofilm morphology and bacterial viability, respectively. When biofilms were exposed to twice-daily blue light, EPS-insoluble was reduced significantly more than in either control group (CHX and 0.89% NaCl). Bacterial viability and dry weight were also reduced relative to the negative control (0.89% NaCl) when the biofilms were treated with twice-daily blue light. Different morphology was also visible when the biofilms were treated with blue light., Conclusions: Twice-daily treatment with blue light without a photosensitizer is a promising mechanism for the inhibition of matrix-rich biofilm development.

Aim: This in situ study assessed the effects of adhesive systems containing or not fluoride and/or the antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB) on the microbiological composition of dental biofilm and enamel demineralization., Materials and Methods: During two phases of 14 days, ten volunteers wore intraoral palatal appliances containing two slabs of human enamel according to a double-blind, crossover design. The slabs were randomly restored using a composite resin and one of the following adhesive systems: All-Bond SE(TM) (self-etch, fluoride/MDPB free adhesive, AB) and Clearfl Protect Bond (self-etch containing fluoride and MDPB adhesive, CB). The biofilm formed on the slabs was analyzed with regard to total and mutans streptococci and lactobacilli counts. Demineralization represented by integrated area of hardness × lesion depth Delta S ( ΔS) was determined on enamel by analysis of cross-sectional microhardness, at 20 and 70 μm from the restoration margin. Data were analyzed by ANOVA., Results: No statistically significant difference was found either in enamel demineralization or in the microbiological composition of dental biofilm., Conclusion: All adhesive systems containing or not fluoride and/or MDPB tested were unable to inhibit secondary caries in the in situ model used in the present research.

This study utilized the confocal Raman microspectroscopy (CRM) technique for the first time to investigate the degree of the penetration of toluidine blue-orto (TBO) in artificial caries lesions produced by two distinct caries-inducing models. The dentin specimens (n = 10) were divided into three groups: control, in vitro and in situ. Thereafter, the lesion depth and the demineralization level were evaluated by cross-sectional microhardness (CSMH). CRM mapping across the dentin surface was assessed after the dye application. The CSMH and CRM data were analyzed by t-test and ANOVA, respectively (P < 0.05). The values of the lesion depth and the demineralization areas were higher for in situ samples (P < 0.05). The TBO penetration values (μm) for the control, in vitro and in situ groups were 44.8 ± 5.6, 46.1 ± 4.5 and 51.2 ± 8.5, respectively. There were no statistically significant differences among the groups (P > 0.05). The rate of TBO penetration was detected up to about <50 μm and the demineralization level did not influence the results. These results have showed promising parameters to develop new protocols for deep caries lesions management using photodynamic antimicrobial chemotherapy., (© 2013 The American Society of Photobiology.)

Unlabelled: Abstract Objective: The aim of the present study was to examine the size and shape of Streptococcus mutans bacterial cells of infected dentin substrate subjected to photodynamic therapy (PDT) using atomic force microscopy (AFM)., Background Data: New trends in the application of AFM have been developed in the field of dentistry, making AFM a useful technique in high resolution imaging of biological structures and processes., Materials and Methods: PDT was completed using an efficient light-emitting diode source (LED - λ=620-660 nm) with total light dose of 94 J/cm(2) in the presence of the photosensitizer toluidine blue O (TBO). Dentin specimens were immersed in brain heart infusion (BHI) broth inoculated with S. mutans for 5 days to induce caries in vitro. After demineralization, the samples were subjected to a series of treatments in which carious dentin infected by S. mutans was exposed to 0.9% sodium chlorite (NaCl) solution (control) for 10 min, or subjected to PDT-TBO photosensitizer followed by light exposure (energy density of 94 J/cm(2))., Results: Three-dimensional (3-D) images and cross-sectional measurements showed rod and diplococcic cell shapes. Photoinactivated bacterial cells did not differ from the control with respect to their cross-sectional shape, but they did show a reduction in size., Conclusions: Photodynamic therapy decreased the diameter of S. mutans cells and AFM may be used as a technique for bacterial cell analysis.

Unlabelled: Several photosensitizers have been used against oral bacteria without standardization. Singlet oxygen ((1)O(2)) is an aggressive chemical species that can kill cells through apoptosis or necrosis., Objective: to compare the antimicrobial activity of photodynamic therapy (PDT) with different photosensitizers at the same concentration against Streptococcus mutans. In addition, the (1)O(2) production of each photosensitizer was determined. The photosensitizers (163.5 μM) methylene blue (MB), toluidine blue ortho (TBO) and malachite green (MG) were activated with a light-emitting diode (LED; λ=636 nm), while eosin (EOS), erythrosine (ERI) and rose bengal (RB) were irradiated with a curing light (λ=570 nm). Light sources were operated at 24 J cm(-2). For each photosensitizer, 40 randomized assays (n=10 per condition) were performed under one of the following experimental conditions: no light irradiation or photosensitizer, irradiation only, photosensitizer only or irradiation in the presence of a photosensitizer. After treatment, serial dilutions of S. mutans were seeded onto brain heart infusion agar to determine viability in colony-forming units per milliliter (CFU mL(-1)). Generation of (1)O(2) was analyzed by tryptophan photooxidation, and the decay constant was estimated. Results were analyzed by one-way ANOVA and the Tukey-Kramer test (p<0.05). PDT with irradiation in the presence of the photosensitizers TBO and MG was effective in reducing S. mutans counts by 3 and 1.4 logs, respectively (p<0.01), compared to their respective untreated controls. MB generated 1.3 times more (1)O(2) than TBO, and both produced significantly higher concentrations of singlet oxygen than the other photosensitizers. Since in vitro bulk (1)O(2) production does not indicate that (1)O(2) was generated in the bacterial activity site, the bactericidal action against S. mutans cannot be related to in vitro singlet O(2) generation rate. In vitroS. mutans-experiments demonstrated TBO as the only photosensitizer that effectively reduced 99.9% of these microorganisms., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Aim: The effect of photodynamic antimicrobial chemotherapy (PACT) on oral biofilms was evaluated., Methods: Biofilms formed in vitro were treated with sensitizer (S) and/or light (L) in the test (S+L+) and control (S-L-, S+L-, S-L+) groups. Additionally, 21 volunteers wore intraoral devices and the biofilms formed on each side of the device were referred to as S+L+ or S-L- groups., Results: Significant decreases in the viability of in vitro biofilms were observed after PACT. PACT was ineffective in inhibiting multi-species biofilms formed in situ., Conclusion: PACT was effective on in vitro biofilms and was ineffective on in situ biofilms., (Copyright © 2012 S. Karger AG, Basel.)

Aim: The aim of this study was to evaluate a very short-term in vitro and in situ effect of 2% chlorhexidine-digluconate-based (CHX) cavity cleanser on the disinfection of dentin demineralized by cariogenic bacteria., Materials and Methods: Human dentin slabs were randomly allocated and used in 2 distinct phases, in vitro and in situ, for obtaining demineralized dentin. In vitro, the slabs (n=15) were immersed for 5 days in BHI broth inoculated with Streptococcus mutans CTT 3440. In situ, a double-blind design was conducted in one phase of 14 days, during which 20 volunteers wore palatal devices containing two human dental dentin slabs. On 5(th) day in vitro and 14(th) day in situ, the slabs were allocated to the two groups: Control group (5 μl of 0.9% NaCl solution) and CHX group (5 μl of 2% chlorhexidine digluconate solution, Cavity Cleanser™ BISCO, Schaumburg, IL, EUA), for 5 minutes. The microbiological analyses were performed immediately before and after the treatments., Results: The log reductions means found for CHX treatment on tested micro organisms were higher when compared to Control group either in vitro or in situ conditions., Conclusions: Our results showed that CHX was effective in reducing the cultivable microbiota in contaminated dentin. Furthermore, although the use of chlorhexidine-digluconate-based cavity disinfectant did not completely eliminate the viable microorganisms, it served as a suitable agent to disinfect tooth preparations.

Objective: The aim of this multidisciplinary study was to evaluate quantitatively and qualitatively the effect of a 660-nm diode laser in the prevention and treatment of human oral mucositis (OM) in patients suffering from head and neck cancer who had undergone radiotherapy and chemotherapy., Background Data: OM is a severe oral lesion resulting from the toxic effects of treatment for cancer in the head and neck region. Low-level laser therapy is indicated to prevent and treat this oral complication and may be used alone or in association with conventional drug treatment, producing pain relief and wound repair., Methods: This study included 72 patients with head and neck cancer treated at the Cancer Hospital of Mato-Grosso, Brazil, and divided into a control group (C; n = 36) and a laser group (L; n = 36). Laser therapy was performed in combination with radiotherapy and chemotherapy twice a week using a diode laser (lambda = 660 nm, power = 30 mW, spot size = 2 mm, energy = 2 J per point)., Results: Statistically significant differences were observed between the two groups. Patients in group L usually did not present with OM or pain, but all patients in group C presented with OM ranging from Level I to III associated with pain. This difference was significant from week 1 on, increased until week 4 and remained stable up to week 7., Conclusion: Laser therapy was effective in preventing and treating oral effects induced by radiotherapy and chemotherapy, thus improving the patient's quality of life.

Photodynamic antimicrobial therapy (PACT) promotes bacterial death as a result of the photosensitization of microbial components. This study evaluated the effect of PACT on dentine caries produced in situ. Over the course of 14 d, 20 volunteers wore intra-oral devices containing human dentine slabs that were treated 10 times daily with a 40% sucrose solution. Afterwards, the antimicrobial effect of toluidine blue O, associated with 47 or 94 J cm(-2) of a light-emitting diode, was evaluated. Before and after the treatments, dentine samples were analysed with regard to the total number of microorganisms, total streptococci, mutans streptococci, and lactobacilli. Significant reductions in the bacterial count were observed for PACT with both energy densities tested, with the following values observed for 47 and 94 J cm(-2) of irradiation: for total streptococci, 3.45 and 5.18; for mutans streptococci, 3.08 and 4.16; for lactobacilli, 3.24 and 4.66; and for total microorganisms, 4.29 and 5.43, respectively. The control, treated with 94 J cm(-2) of irradiation alone, was also effective against all bacteria. To conclude, PACT was effective in killing oral microorganisms present in dentine caries produced in situ and may be a useful technique for eliminating bacteria from dentine carious lesions before restoration.

Background: The purpose of this study was to evaluate the adjunctive clinical, microbiologic, and immunologic effects of the systemic administration of amoxicillin and metronidazole in the full-mouth ultrasonic debridement of patients with severe chronic periodontitis., Methods: Twenty-five patients presenting at least eight teeth with probing depth (PD) > or =5 mm and bleeding on probing (BOP) were selected and randomly assigned to full-mouth ultrasonic debridement + placebo (control group) or full-mouth ultrasonic debridement + amoxicillin and metronidazole (test group). The clinical outcomes evaluated were visible plaque index, BOP, position of the gingival margin, relative attachment level (RAL), and PD. Real-time polymerase chain reaction (PCR) was used for quantitative analysis of Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans), Porphyromonas gingivalis, and Tannerella forsythia (previously T. forsythensis). The enzyme-linked immunosorbent assay (ELISA) technique permitted the detection of prostaglandin E(2,) interleukin-1beta, and interferon-gamma levels in gingival crevicular fluid. All parameters were evaluated at baseline and at 3 and 6 months post-treatment., Results: At 6 months, the test treatment resulted in lower BOP and an additional reduction (0.83 mm) in PD (P <0.05). Data also showed RAL gain > or =2 mm at 43.52% of sites in control patients compared to 58.03% of sites in test patients (P <0.05). However, both groups had similar mean RAL gain (1.68 and 1.88 mm for the control and test groups, respectively). Real-time PCR and ELISA failed to identify significant differences between the groups., Conclusions: Both treatments resulted in significant clinical improvements; however, there was a slight, but significantly greater, improvement in BOP and the percentage of sites with PD > or =5 mm exhibiting RAL gain > or =2 mm in the test group. Nevertheless, no improvement in the microbiologic or immunologic outcome was observed with the adjunctive use of systemic amoxicillin and metronidazole.

Objectives: Since secondary caries is one of the main reasons for replacing restorations, this study assessed the effects of different restorative materials on the microbiological composition of dental biofilm and on enamel demineralisation around the restoration., Methods: A randomized, double-blind, split-mouth in situ design was conducted in one phase of 14 days, during which, 20 volunteers wore palatal devices containing five human dental enamel slabs. Each slab was randomly restored with one of the following materials: Filtek-Z-250/Single Bond, control group (composite resin), Permite (amalgam), Fuji II (encapsulated resin-modified glass ionomer), Vitremer (resin-modified glass ionomer) and Ketac Molar (conventional glass ionomer). The volunteers used fluoride dentifrice, 3x/day and a 20% sucrose solution was dripped onto the slabs 8x/day. The biofilm formed on the slabs was analyzed to determine the counts of total streptococci, mutans streptococci and lactobacilli. Enamel demineralisation was determined by cross-sectional microhardness (CSMH) at 20 and 70 microm from the margin of the restoration. Kruskal-Wallis and analysis of variance, followed by least mean squares (LMS) test, were used to evaluate microbiota and CSMH among the groups. The significance level used was 5%., Results: No statistically significant differences were found in the cariogenic microbiota grown on the slabs. At a 20-mum distance, only Fuji II statistically differed from the other groups, showing the lowest demineralisation. At 70 microm, Fuji II significantly inhibited demineralisation when compared to Permite, Filtek-Z-250 and Ketac Molar., Conclusions: In the context of fluoride dentifrice and under the cariogenic exposure conditions of this study, only the encapsulated resin-modified glass ionomer material provided additional protection against secondary caries.

Aim: Causes of temporomandibular disorders (TMDs) are multifactorial, involving both physical and psychological factors. The aim of this study was to evaluate the salivary cortisol levels in young adults with TMDs., Methods: The Research Diagnostic Criteria (RDC)/TMDs were used to establish the diagnosis of TMDs regarding the physical (axis I) and psychological factors (depression and somatization; axis II). Young adults of both genders composed the TMDs males (21.7+/-2.21 years, n=10) and females (22.7+/-1.95 years n=10) groups, and 20 healthy matched controls composed the control male (22.5+/-1.96 years n=10) and female (22.4+/-2.17years n=10) groups. Saliva samples were collected immediately on awakening and at bedtime. The cortisol levels were measured by enzyme immunoassay., Results: The female group with TMDs showed a significantly higher cortisol level (2.890+/-0.411 mg/dL) than the corresponding control group in the morning evaluation (2.470+/- 0.486 mg/dL). The depression scores were significantly higher for the female group with TMDs (1.3+/-0.18) in comparison with the control female (0.72+/-0.18) and male groups (0.51+/-0.13). The groups with TMDs showed significantly higher somatization scores (0.69+/-0.16 for males and 1.38+/-0.23 for females) in comparison with the controls (0.23+/-0.10 for males and 0.78+/-0.15 for females), these also being significantly higher for the females than males., Conclusion: It was concluded that cortisol levels are higher in TMDs in females, who had elevated depression and somatization scores. The results infer that the evaluation of salivary cortisol could be a promising tool in the diagnosis of TMDs associated with psychological factors.

In natural ecosystems, micro-organisms grow preferentially attached to surfaces, forming matrix-enclosed biofilms. The aim of this study was to determine photodestruction levels in biofilms after subjecting them to photodynamic therapy. Biofilms of Streptococcus mutans, S. sobrinus, and S. sanguinis were grown on enamel slabs for 3, 5 or 7 d. Both the number of viable micro-organisms and the concentration of water-insoluble polysaccharides were analysed, and mineral loss (DeltaZ) analyses were performed on the enamel slabs. The antimicrobial potential of toluidine blue O (0.1 mg ml(-1)), associated with 85.7 J cm(-2) of a light-emission diode, was evaluated on the viability of 5-d biofilms. Both the number of micro-organisms and the concentration of water-insoluble polysaccharide increased with the age of the biofilms. A significant reduction ( approximately 95%) in viability was observed for S. mutans and S. sobrinus biofilms following photosensitization, with a > 99.9% reduction in the viability of S. sanguinis biofilms. In conclusion, a biofilm model was shown to be suitable for studying changes in bacterial numbers and enamel mineralization and for demonstrating the potential value of photosensitization in the control of in vitro biofilms.

Objectives: The purpose of this study was to evaluate the antimicrobial effect of toluidine blue O (TBO), in combination with either a helium/neon (HeNe) laser or a light-emitting diode (LED), on the viability and architecture of Streptococcus mutans biofilms., Methods: Biofilms were grown on hydroxyapatite discs in a constant depth film fermentor fed with artificial saliva that was supplemented with 2% sucrose four times a day, thus producing a typical 'Stephan pH curve'. Photodynamic therapy was subsequently carried out on biofilms of various ages with light from either the HeNe laser or LED using energy densities of between 49 and 294 J/cm(2)., Results: Significant decreases in the viability of S. mutans biofilms were only observed when biofilms were exposed to both TBO and light, when reductions in viability of up to 99.99% were observed with both light sources. Overall, the results showed that the bactericidal effect was light dose-dependent and that older biofilms were less susceptible to photodynamic therapy. Confocal laser scanning microscopy images suggested that lethal photosensitization occurred predominantly in the outermost layers of the biofilms., Conclusions: Photodynamic therapy may be a useful approach in the treatment of dental plaque-related diseases.

Objective: The purpose of this study was to evaluate the effect of erbium:yttrium-aluminum-garnet laser instrumentation of root surfaces on the morphology of fibroblasts from continuous lineage., Method and Materials: Dentinal slices with 4 mm2 of surface area were obtained from teeth extracted for severe periodontal involvement. Specimens were assigned to one of three treatment groups: group 1, application of the laser with an energy level of 250 mJ at 103 pulses per second; group 2, application of the laser with an energy level of 80 mJ at 166 pulses per second; and group 3, similar to group 2, but with concomitant water irrigation of the device. The specimens were incubated in multiwell plates containing cell culture media. After 24 hours, the specimens were submitted to routine preparation for scanning electron microscopy. Three independent and blind examiners used photomicrographs to evaluate the morphology of the fibroblasts: 0 = without cells; 1 = flat cells; 2 = round cells; and 3 = combination of round and flat cells., Results: Statistical analysis indicated that there were significant differences among treatment groups and that group 3 was significantly different from groups 1 and 2., Conclusion: There was no difference between groups 1 and 2 in the morphology of fibroblasts. Laser instrumentation with concomitant irrigation impaired the adhesion of fibroblasts to dentinal surfaces.