Combining Coaxial Electrospinning and 3D Printing: Design of Biodegradable Bilayered Membranes with Dual Drug Delivery Capability for Periodontitis Treatment.

Periodontitis is a chronic inflammatory disease that can lead to significant destruction of tooth-supporting tissues, compromising dental function and patient's health. Although the currently employed treatment approaches can limit the advance of the disease, the development of multifunctional and hierarchically structured materials is still in demand for achieving successful tissue regeneration. Here, we combine coaxial electrospinning and 3D printing techniques to prepare bilayered zein-based membranes as a potential dual drug delivery platform for periodontal tissue regeneration. A layer of core-sheath electrospun nanofibers consisting of poly(ethylene oxide) (PEO)/curcumin (Curc)/tetracycline hydrochloride (TH) as the core and zein/poly(ε-caprolactone)(PCL)/ß-glycerolphosphate (ß-GP) as the sheath was deposited over a 3D printed honeycomb PLA/zein/Curc platform in order to render a bilayered structure that can mimic the architecture of periodontal tissue. The physicochemical properties of engineered constructs as well as the release profiles of distinct drugs were mainly controlled by varying the concentration of zein (10, 20, 30%, w/w relative to dry PCL) on the sheath layer of nanofibers, which displayed average diameters ranging from 150 to 400 nm. In vitro experiments demonstrated that the bilayered constructs provided sustained release of distinct drugs over 8 days and exhibited biocompatibility toward human oral keratinocytes (Nok-si) (cell viability >80%) as well as antibacterial activity against distinct bacterial strains including those of the red complex such as Porphyromonas gingivalis and Treponema denticola, which are recognized to elicit aggressive and chronic periodontitis. Our study reveals the potential of zein-based bilayered membranes as a dual drug delivery platform for periodontal tissue regeneration.

A quest to find good primers for gene expression analysis of Candida albicans from clinical samples.

Biofilm production contributes to several human diseases, including oral candidiasis. Among the Candida species, Candida albicans is the most prevalent. The expression of virulence genes is implicated in the pathogenic potential of Candida biofilms. However, the evaluation of microbial gene expression from in vivo biofilm samples is not trivial, specifically, assessment via quantitative PCR (qPCR) can be a challenge because of several species present in clinical samples. Hence, the necessity of primers specificity. The aim of this study was to evaluate through in silico and in vitro analyses the specificity of published primers and newly designed primers for C. albicans virulence genes: ALS1, CAP1, CAT1, EFG1, HWP1, LIP3, PLB1, SAP1, SAP4, SOD1, SOD5 and ACT1 (normalizing gene). In silico analysis was performed through a PubMed search of articles with primer sequences that evaluated gene expression of C. albicans. Then, the sequence similarity of twenty-eight primers was checked through BLASTn and ClustalW2. The analysis of secondary structures was performed using mfold. When the primers did not present satisfactory characteristics (absence of secondary structures, not discrepant Tm of forward and reverse sequences and specificity) following in vitro analysis (i.e., end point PCR), new primers were designed using Beacon Designer™ and sequences obtained from the "Candida Genome Database". The selected primers were tested in vitro by end point PCR using a panel of genomic DNA from five different Candida species (C. albicans, Candida glabrata, Candida dubliniensis, Candida krusei, and Candida tropicalis). The resulting PCR products were visualized on agarose gel. qPCR reactions were performed to determine primers' optimal concentration and PCR efficiency. End point PCR demonstrated that published primers for the SAP1 and HWP1 were specific for C. albicans and the one for SOD1 reacted with C. albicans and C. dubliniensis. The sequence of primers designed for ACT1, ALS1 and HWP1 genes were specific for C. albicans, while the ones for CAP1, CAT1, EFG1, LIP3, and PLB1 were detected in C. albicans and C. dubliniensis. After optimization, all primers presented a single peak on melt curves, correlation coefficient of ≅1 and qPCR reaction efficiency of 90-110%, with slope of ≅-3.3. Therefore, these primers should be suitable for future gene expression analyses from clinical samples.

A Survey of the Management of Patients with Temporomandibular Disorders by General Dental Practitioners in Southern Brazil.

PURPOSE: Temporomandibular disorders (TMD) are recognized as one of the most controversial topics in dentistry, despite the fact that both basic science and clinical researchers have currently reached some degree of consensus. This study aimed to conduct a questionnaire-based survey about the management of TMD patients by general dental practitioners (GDPs). MATERIALS AND METHODS: One hundred fifty-one GDPs with a private practice in a city of southern Brazil were included, independent of school of origin, gender, graduation year, and curriculum content. All participants were administered a questionnaire about the management of patients with TMD, and the responses were analyzed by binomial and chi-square tests (α = 0.05). RESULTS: Of the GDPs, 88.7% received TMD patients, who were primarily diagnosed on the basis of medical history (36.6%) or physical examination (30.4%). Of these, 65.4% referred the patients elsewhere, primarily to specialists in occlusion (36.1%) or orthodontics (29.7%). Occlusal splinting was the most commonly used management modality (20.8%), followed by occlusal adjustment (18.1%) and pharmacotherapy (16.6%). Splints were fabricated in maximum habitual intercuspation or centric relation depending on individual patient (54.8%). The hard stabilization form was the most common type of appliance used (35.0%). Moreover, 73.8% of the GDPs did not employ semi-adjustable articulators, and 69.5% adjusted the appliances at the time of fixing. The duration of splint use and the frequency of follow-up were considered patient dependent by 62.1% and 72.8%, respectively. GDPs considered the two major TMD etiologic categories as multifactorial (20.8%) and occlusion (19.9%). Multidisciplinary medical and dental treatment was considered necessary by 97.9%. CONCLUSIONS: The evaluated general dental practitioners manage TMD patients according to international guidelines.

Surface roughness and Candida albicans biofilm formation on a reline resin after long-term chemical disinfection and toothbrushing.

STATEMENT OF PROBLEM: Routine cleaning of a denture may increase the surface roughness of the material. PURPOSE: The purpose of this study was to investigate the effect of denture cleansers and time on the roughness and Candida albicans biofilm formation on a reline resin. MATERIAL AND METHODS: Specimens of Tokuyama Rebase Fast II (20 × 10 × 2 mm) were prepared and divided into 9 groups (n=15): Test groups were toothbrushed (30 cycles per day) in 1 of the following solutions: water, soap, or toothpaste. After toothbrushing, the specimens were immersed in solutions of water, sodium perborate, or chlorhexidine. These treatments were done once a day for 365 days. The surface roughness was evaluated at 0, 7, 15, 30, 90, 180, 270, and 365 days, and the C albicans biofilm formation was measured after 365 days. The roughness data were analyzed by 3-way repeated measures ANOVA and the Tukey test (α=.05), and the C albicans biofilm formation was analyzed by 2-way ANOVA. RESULTS: Significant differences were found in the toothbrush and time interaction and in the toothbrush, immersion, and time interaction (P<.001). No significant differences were found between the toothbrush agent and chemical disinfection (P=.085) or between chemical disinfection and time interaction (P=.604). Brushing with dentifrice (PPb and PCh) showed a gradual decrease in surface roughness. The comparison among groups showed that PCh produced the lowest values of roughness. For C albicans biofilm formation, no significant differences were found among the experimental groups. CONCLUSIONS: The roughness values ranged from 0.31 to 0.69 µm for all evaluated groups. For all groups, no significant differences were found in the quantification of C albicans.

Effect of human whole saliva on the in vitro adhesion of Candida albicans to a denture base acrylic resin: a focus on collection and preparation of saliva samples.

OBJECTIVE: In studies on Candida albicans adhesion to surfaces, diverse protocols have been used for collection and preparation of saliva samples. Thus, this study investigated whether variations in the centrifugation parameters and number of donors of saliva would influence the adhesion of C. albicans to a denture base resin. METHODS: Resin acrylic samples (n = 72) were made and then divided into four groups: (a) control - specimens were left without preconditioning in saliva; (b) three experimental groups, in which the specimens were preconditioned with saliva collected from 15 volunteers and centrifuged at 12 000 g for 5 min (G1 ); from 15 volunteers and centrifuged at 18 000 g for 30 min (G2 ); and from one volunteer and centrifuged at 12 000 g for 5 min (G3 ). Candida adhesion was evaluated by both the 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide (XTT) reduction method and crystal violet staining. Data were analyzed by one-way analyses of variance (P = 0.05). RESULTS: For XTT reduction assay, groups G2 , G3 , and control were not significantly different, whereas group G1 showed significantly higher absorbance value than control. For crystal violet staining there were no significant differences among all groups. CONCLUSION: Variations in the centrifugation parameters and number of donors of saliva may influence C. albicans adhesion to denture base resins.

Clinical evaluation of failures in removable partial dentures.

The aim of this clinical study was to evaluate the effects of removable partial dentures on the support tissues and changes occurring in lower tooth-supported and bilateral distal-extension dentures, 5 years after placement. The study involved analysis of a total of 53 patients who received prosthetic treatment for removable partial dentures. The patients were divided into two groups. In group 1, the patients had a completely edentulous maxilla and an edentulous area with natural teeth remaining in both the anterior and posterior regions. In group 2, the patients had a completely edentulous maxilla and partially edentulous mandible with preserved anterior teeth. Tooth mobility, prevalence of caries, fracture of the abutment teeth, fracture and/or deformation of the removable partial denture components and stability of the denture base were evaluated. The use of a removable partial denture increased tooth mobility, reduced the prevalence of caries, and did not cause loss or fracture of the abutments or damage to their components, when compared with the baseline. It was concluded that there was no difference between the groups as evaluated in terms of tooth mobility, prevalence of caries, loss and fracture of the abutments or damage to the components of the removable partial denture.

Prevalence of Candida spp. associated with bacteria species on complete dentures.

OBJECTIVE: This study evaluated the prevalence of Candida spp., Streptococcus mutans and Staphylococcus aureus isolated from complete upper dentures. BACKGROUND: Factors that may increase the number of microorganisms in the oral cavity include denture-wearing and deficient denture hygiene. MATERIALS AND METHODS: Ninety denture wearers participated in this study. Collection of biological material from each denture was performed on all denture surfaces. Conventional biochemical methods were used to identify the microorganisms. Descriptive statistical analysis was performed on percentage data. RESULTS: Candida spp. were the most frequent microorganisms found on dentures (65.5%), while Strep. mutans and Staph. aureus were present in 53.3 and 34.4%, respectively. CONCLUSION: Candida species were the most frequent microorganisms on dentures, followed by Strep. mutans and Staph. aureus.

Evaluation of the occlusion vertical dimension of complete dentures after microwave disinfection.

OBJECTIVES: An increase in occlusal vertical dimension (OVD) after microwave disinfection may result in the need for adjustments in the complete dentures. This in vitro study evaluated the increase in OVD of maxillary complete dentures submitted to microwave disinfection protocols. MATERIAL AND METHODS: Thirty sets of complete dentures were evaluated as follows: Group 1-15 sets had the maxillary complete dentures submitted to microwave disinfection (650 W/3 min), once a week, for 4 weeks. Group 2-15 sets had the maxillary complete dentures submitted to microwave disinfection (650 W/3 min), three times a week, for 4 weeks. The vertical dimension was measured with a micrometre (in mm) before disinfection protocols (baseline readings) and after each week of disinfection. Data were analysed using Wilcoxon and Friedman tests (α = 0.05). RESULTS: For Group 1, no significant difference was found between the increases in OVD and zero, and no significant difference was found between the weeks. For Group 2, the increases in OVD were significantly greater than zero, and the Friedman test showed that weeks 3 and 4 had significantly greater changes than week 1 and that week 4 had significantly greater change than week 2. CONCLUSION: Microwave disinfection only promoted significant increase in OVD in Group 2, in which the values increased progressively.

Susceptibility of clinical isolates of Candida to photodynamic effects of curcumin.

BACKGROUND AND OBJECTIVE: The resistance of Candida species to antifungals represents a major challenge for therapeutic and prophylactic strategies. This study evaluated photodynamic therapy (PDT) mediated by Curcumin (CUR) against clinical isolates of C. albicans, C. tropicalis, and C. glabrata, both in planktonic and biofilm forms. STUDY DESIGN/MATERIALS AND METHODS: Suspensions of Candida were treated with three CUR concentrations and exposed to four LED fluences. The protocol that showed the best outcomes for inactivation of the planktonic phase was selected to be evaluated against Candida biofilms. In addition, two higher CUR concentrations were tested. The metabolic activity of biofilms was evaluated by means of XTT reduction assay and the biofilm biomass was evaluated using crystal violet (CV) staining assay. Data were analyzed in a mixed model nested ANOVA, Wilcoxon's nonparametric tests, and the Kruskal-Wallis test (α = 5%). RESULTS: The use of CUR in association with light was able to promote a significant antifungal effect against the planktonic form of the yeasts. When using 40 µM of CUR, the metabolic activity of C. albicans, C. glabrata, and C. tropicalis biofilms was reduced by 85%, 85%, and 73%, respectively, at 18 J/cm(2) . CUR-mediated PDT also decreased the biofilm biomass of all species evaluated. In addition, CV staining showed that C. albicans isolates were strong biofilm-forming strains, when compared with C. glabrata and C. tropicalis isolates. CONCLUSION: The results from the present investigation showed that low CUR concentrations can be highly effective for inactivating Candida isolates when associated with light excitation.

Effect of water-bath post-polymerization on the mechanical properties, degree of conversion, and leaching of residual compounds of hard chairside reline resins.

OBJECTIVES: This study evaluated the effect of water-bath post-polymerization at 55 degrees C for 10 min (WB) on the content and leaching of residual compounds, degree of conversion, flexural strength, and hardness of hard chairside reline resins (Kooliner: K, New Truliner: N, Ufi Gel hard: U, and Tokuso Rebase Fast: T). METHODS: Leaching experiments were made by storing specimens (n=48) in artificial saliva at 37+/-1 degrees C and analyzing residual monomers and plasticizer by HPLC. Analysis of residual monomer and plasticizer content (n=48) was also made by HPLC. Degree of conversion (n=40) was analyzed by using FT-Raman spectroscopy. A 3-point loading test was used to evaluate the flexural strength of the specimens (n=80). One fragment of each flexural test specimen was then submitted to Vickers microhardness test. RESULTS: WB produced a significant decrease (p<0.050) in the amount of residual compounds eluted from the materials within the first hour of immersion. With the exception of material U, WB decreased the duration of release of the residual compounds evaluated. All materials evaluated exhibited significantly (p<0.050) lower values of residual monomer and plasticizer (material N) after WB compared with the control groups. WB increased the degree of conversion of K and T resins and the hardness of N, K, and T resins (p<0.050). Only material K showed an increase in flexural strength after WB (p<0.050). SIGNIFICANCE: Immersion of relined dentures in water at 55 degrees C for 10 min can be used to reduce the amounts and duration of release of residual compounds and improve mechanical properties of some of the materials evaluated.

Residual monomer of reline acrylic resins. Effect of water-bath and microwave post-polymerization treatments.

OBJECTIVES: This study compared the residual monomer (RM) in four hard chair-side reline resins (Duraliner II-D, Kooliner-K, Tokuso Rebase Fast-TRF and Ufi Gel hard-UGH) and one heat-polymerized denture base resin (Lucitone 550-L), which was processed using two polymerization cycles (short-LS and long-LL). It was also investigated the effect of two after polymerization treatments on this RM content. METHODS: Specimens (n=18) of each material were produced following the manufacturers' instructions and then divided into three groups. Group I specimens were left untreated (GI-control). Specimens of group II (GII) were given post-polymerization treatment by microwave irradiation. In group III (GIII), specimens were submitted to immersion in water at 55 degrees C (reline resins-10 min; denture base resin L-60 min). The RM was analyzed using high performance liquid chromatography (HPLC) and expressed as a percentage of RM. Data were analyzed by two-way ANOVA followed by Tukey's test (alpha=0.05). RESULTS: Comparing control specimens, statistical differences were found among all materials (p<0.05), and the results can be arranged as K (1.52%)>D (0.85%)>UGH (0.45%)>LL (0.24%)>TRF (0.14%)>LS (0.08%). Immersion in hot water (GIII) promoted a significant (p<0.05) reduction in the RM for all materials evaluated compared to control (GI), with the exception of LL specimens. Materials K, UGH and TRF exhibited significantly (p<0.05) lower values of RM after microwave irradiation (GII) than in the control specimens. SIGNIFICANCE: The reduction in RM promoted by water-bath and microwave post-polymerization treatments could improve the mechanical properties and biocompatibility of the relining and denture base materials.