Resumo em Inglês:

Objective : The aim of this study was to review the reliability and reproducibility of 3D-CBCT (cone beam computed tomography) cephalometric landmark identification. Methods : Electronic databases (Pubmed, Scopus, Web of Science) were searched for papers published from 1998 to October 2014. Specific strategies were developed for each database, with the guidance of a librarian. Two reviewers independently analyzed the titles and abstracts for inclusion. The articles that met the inclusion and exclusion criteria were selected for full-text reading, and the selected articles went through methodological quality evaluation. After the exclusion of repeated articles, the titles of the remaining ones were read and 1,328 of them were excluded. The abstracts of 173 articles were read, of which 43 were selected, read in full and submitted to the inclusion and exclusion criteria. Fourteen articles or studies with reliable methodology and reproducibility remained. The data were collected, organized into figures and analyzed for determination of the reliability and reproducibility of the three-dimensional cephalometric landmarks. Results : Overall, the landmarks on the median sagittal line and dental landmarks had the highest reliability, while the landmarks on the condyle, porion and the orbitale presented lower levels of reliability. Point S must be marked in the multiplanar views associated with visualization in 3D reconstruction. Further studies are necessary for evaluating soft tissue landmarks.

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Objective: To investigate the mechanical properties of different classifications of composites indicated for posterior application as functions of the storage condition and of the energy dose. Material and Methods: Specimens (8x2x2 mm) were obtained according to the factors: I) Composites (3M ESPE): Filtek P60, Filtek Z350XT, and Filtek Silorane; II) Syringe storage conditions: room temperature, aged, oven, refrigerator, and freezer; and III) Energy dose: 24 J/cm2 and 48 J/cm2. After photoactivation, the specimens were stored at 37ºC for 24 h. After storage, a three-point bending test was carried out in a universal testing machine at 0.5 mm/min. Flexural strength (S) and flexural modulus (E) were calculated. Data were analyzed by three-way ANOVA and Tukey's test (α=0.05). Results: Different storage conditions significantly affected the silorane composite for S; conversely, no effects were noted in terms of E. The accelerated aging protocol significantly increased the S of Filtek P60 and Filtek Silorane, whereas storage in the oven significantly decreased the S for all of the composites tested. Filtek P60 was the only composite not affected by the lower storage temperatures tested for S, whereas for the silorane this parameter was impacted at the same conditions. The factor "dose" was not statistically significant. Conclusions: The syringe storage at different temperature conditions proved to influence mostly the flexural strength, a clinically important characteristic considering the posterior indication of the materials tested. The silorane composite should not be stored at lower temperatures.

Resumo em Inglês:

OBJECTIVES: This cross-sectional study aimed to evaluate the influence of Primary Headache (PH) on efficacy of a Temporomandibular Disorders (TMD) conservative therapy and its association with the presence of self-reported parafunctional habits. SAMPLE AND METHODS: Sample was composed of 400 medical records, divided into four groups: I) Muscular TMD (n=64); II) Muscular TMD+PH (n=48); III) Muscular TMD+Articular TMD (n=173); IV) Muscular TMD+Articular TMD+PH (n=115). All groups had undergone a TMD therapy for three months with a stabilization appliance and counseling for habits and behavioral changes, with no specific headache management. Current pain intensity and existence or not of self-reported bruxism were assessed. Repeated measures ANOVA and Chi-Square test followed by Odds were used for statistical analysis, with a significance level of 5%. RESULTS: results of this study showed that: (1) A conservative therapy with stabilization appliance and counseling for habits and behavioral changes was effective in the TMD pain relief; (2) Groups with an additional diagnosis of PH had worsened the pain improvement significantly; and (3) no association between the presence of self-reported bruxism and PH was found. CONCLUSIONS: this study could elucidate the important effect that headache may have on the TMD management.

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The mandible condylar process cartilage (CP) of Wistar rats is a secondary cartilage and acts as a mandibular growth site. This phenomenon depends on adequate proteins intake and hormone actions, including insulin. Objectives The present study evaluated the morphological aspects and the expression of the insulin receptor (IR) in the cartilage of the condylar process (CP) of rats subjected to protein undernourishment. Material and Methods The nourished group received a 20% casein diet, while the undernourished group (U) received a 5% casein diet. The re-nourished groups, R and RR, were used to assess the effects of re-nutrition during puberty and adulthood, respectively. CPs were processed and stained with picro-sirius red, safranin-O and azocarmine. Scanning electron microscopy and immunohistochemistry were also performed. Results The area of the CP cartilage and the number of cells in the chondroblastic layer decreased in the U group, as did the thickness of the CP layer in the joint and hypertrophic layer. Renourishment during the pubertal stage, but not during the adult phase, restored these parameters. The cell number was restored when re-nutrition occurred in the pubertal stage, but not in the adult phase. The extracellular matrix also decreased in the U group, but was restored by re-nutrition during the pubertal stage and further increased in the adult phase. IR expression was observed in all CPs, being higher in the chondroblastic and hypertrophic cartilage layers. The lowest expression was found in the U and RR groups. Conclusions Protein malnutrition altered the cellularity, the area, and the fibrous cartilage complex, as well as the expression of the IRs.

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Periodontal ligament mesenchymal stem cells (PDLMSCs) are an important alternative source of adult stem cells and may be applied for periodontal tissue regeneration, neuroregenerative medicine, and heart valve tissue engineering. However, little is known about the impact of bacterial toxins on the biological properties of PDLSMSCs, including self-renewal, differentiation, and synthesis of extracellular matrix. Objective : This study investigated whether proliferation, expression of pro-inflammatory cytokines, and osteogenic differentiation of CD105-enriched PDL progenitor cell populations (PDL-CD105+ cells) would be affected by exposure to bacterial lipopolysaccharide fromEscherichia coli (EcLPS). Material and Methods : Toll-like receptor 4 (TLR4) expression was assessed in PDL-CD105+ cells by the immunostaining technique and confirmed using Western blotting assay. Afterwards, these cells were exposed to EcLPS, and the following assays were carried out: (i) cell viability using MTS; (ii) expression of the interleukin-1 beta (IL-1β), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor alpha (TNF-α) genes; (iii) osteoblast differentiation assessed by mineralization in vitro, and by mRNA levels of run-related transcription factor-2 (RUNX2), alkaline phosphatase (ALP) and osteocalcin (OCN) determined by quantitative PCR. Results : PDL-CD105+ cells were identified as positive for TLR4. EcLPS did not affect cell viability, but induced a significant increase of transcripts for IL-6 and IL-8. Under osteogenic condition, PDL-CD105+ cells exposed to EcLPS presented an increase of mineralized matrix deposition and higher RUNX2 and ALP mRNA levels when compared to the control group. Conclusions : These results provide evidence that CD105-enriched PDL progenitor cells are able to adapt to continuous Escherichia coli endotoxin challenge, leading to an upregulation of osteogenic activities.

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Although the interference of tongue-tie with breastfeeding is a controversial subject, The use of lingual frenotomy has been widely indicated by health professionals. Objective : To observe changes in breastfeeding patterns after lingual frenotomy concerning the number of sucks, pause length between groups of sucking and mother's complaints. Material and Methods : Oral yes/no questions about breastfeeding symptoms and sucking/swallowing/breathing coordination were answered by the mothers of 109, 30 day old infants. On the same day the infants had their lingual frenulum assessed by administering a lingual frenulum protocol. After the assessment, all tongue-tied infants were referred for frenotomy; nevertheless, only 14 underwent the surgery. Of the 109 infants, 14 infants who did not have frenulum alterations were included as controls. Birth order and gender were the criteria for recruiting the control group. The tongue-tied infants underwent lingual frenotomy at 45 days of age. At the conclusion of the frenotomy, the infants were breastfed. At 75 days old, both groups – control and post-frenotomy – were reassessed. Before the reassessment the same oral yes/no questions were answered by the mothers of the 14 infants who underwent frenotomy. The mothers of the control group answered the questionnaire only at the time of the first assessment. Data were subjected to statistical analysis. Results : After frenotomy, the number of sucks increased and the pause length between sucking decreased during breastfeeding. The controls maintained the same patterns observed in the first assessment. From the questionnaire answered by the mothers of the 14 tongue-tied infants, at 30 days and 75 days, we observed that the symptoms concerning breastfeeding and sucking/swallowing/breathing coordination were improved after lingual frenotomy Conclusions : after lingual frenotomy, changes were observed in the breastfeeding patterns of the the tongue-tied infants while the control group maintained the same patterns. Moreover, all symptoms reported by the mothers of the tongue-tied infants had improved after frenotomy.

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Phosphoric acid has been suggested as an irrigant due to its effectiveness in removing the smear layer. Objectives : The purpose of this study was to compare the antimicrobial and cytotoxic effects of a 37% phosphoric acid solution to other irrigants commonly used in endodontics. Material and Methods : The substances 37% phosphoric acid, 17% EDTA, 10% citric acid, 2% chlorhexidine (solution and gel), and 5.25% NaOCl were evaluated. The antimicrobial activity was tested against Candida albicans, Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Actinomyces meyeri, Parvimonas micra, Porphyromonas gingivalis, and Prevotella nigrescensaccording to the agar diffusion method. The cytotoxicity of the irrigants was determined by using the MTT assay. Results : Phosphoric acid presented higher antimicrobial activity compared to the other tested irrigants. With regard to the cell viability, this solution showed results similar to those with 5.25% NaOCl and 2% chlorhexidine (gel and solution), whereas 17% EDTA and 10% citric acid showed higher cell viability compared to other irrigants. Conclusion : Phosphoric acid demonstrated higher antimicrobial activity and cytotoxicity similar to that of 5.25% NaOCl and 2% chlorhexidine (gel and solution).

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The search for new instruments to promote an appropriate cervical preparation has led to the development of new rotary instruments such as TripleGates. However, to the best of the authors' knowledge, there is no study evaluating TripleGates effect on the “risk zone” of mandibular molars. Objectives : The aim of this study was to evaluate the effects of a crown-down sequence of Gates-Glidden and TripleGates burs on the remaining cervical dentin thickness and the total amount of dentin removed from the root canals during the instrumentation by using cone beam computed tomography. The number of separated instruments was also evaluated. Material and Methods : Mesial roots of 40 mandibular first molars were divided into 2 equal groups: crown-down sequence of Gates-Glidden (#3, #2, #1) and TripleGates burs. Cervical dentin thickness and canal area were measured before and after instrumentation by using cone beam computed tomography and image analysis software. Student’s t-test was used to determine significant differences at p<0.05. Results : No significant differences (p>0.05) were observed between the instruments, regarding the root canal area and dentin wall thickness. Conclusion : Both tested instruments used for cervical preparation were safe to be used in the mesial root canal of mandibular molars.

Resumo em Inglês:

Oral squamous cell carcinoma (OSCC) is one of the most common malignances. In epithelial-mesenchymal transition (EMT), epithelial cells switch to mesenchymal-like cells exhibiting high mobility. This migratory phenotype is significant during tumor invasion and metastasis. Objective : The aim of this study is to evaluate the expression of the EMT markers E-cadherin, N-cadherin and vimentin in OSCC. Material and Methods : Immunohistochemical detection of E-cadherin, N-cadherin and vimentin was performed on 20 OSCC samples. Differences in the expression of each protein at the invasive front (IF) and in the central/superficial areas (CSA) of the tumor were assessed. Differences in the expression of each protein at the IF of both histologically high- and low-invasive OSCCs were evaluated. Associations among expression of proteins at the IF were assessed. Correlations between the expression levels of each protein at the IF and the tumor stage and clinical nodal status were also evaluated. Results : Reduced expression of E-cadherin was detected in 15 samples (75%). E-cadherin expression was reduced at the IF when compared to the CSA and in high-invasive tumors when compared to low-invasive tumors. All samples were negative for N-cadherin, even though one sample showed an inconspicuous expression. Positive expression of vimentin was observed in 6 samples (30%). Nevertheless, there was no difference in vimentin expression between the IF and the CSA regions or between the low- and high-invasive tumors. Furthermore, no association was observed among protein expression levels at the IF. Finally, no correlations were observed between each protein’s expression levels and tumor stage or clinical nodal status. Conclusions : Reduced E-cadherin expression at the IF and its association with histological invasiveness suggest that this protein is a noteworthy EMT marker in OSCC. Although vimentin was also detected as an EMT marker, its expression was neither limited to the IF nor was it related to histological invasiveness.

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Since the beginning of their lives, all living organisms are exposed to the influence of geomagnetic fields. Objectives : With respect to the positive effects that magnetic fields have on human tissues, especially the bactericidal effect, this investigation aimed to assess their influence on the reduction of oral microorganisms. Material and Methods : In order to obtain adequate specimens of dental plaque deposit, microbes such as Streptococcus parasanguinis, Staphylococcus epidermidis, Rhodococcus equi andCandida albicans were isolated from the human mouth. To establish the intensity of microbial growth on the basis of the modified optical density (OD) of agar turbidimetry assay, microbial count and spectrophotometry were applied. The study was carried out with two microbial concentrations (1 and 10 CFU/ml) after periods of incubation of 24 and 48 h and using micromagnets. Results : A positive effect of the magnetic field, resulting in the reduction of dental plaque microbes in vitro, was found. In the first 24 hours of exposure to the magnetic field, the number of all isolated microbes was significantly reduced. The most potent influence of magnets and the most intensified reduction after 24 hours were evident in Candida albicans colonies. The decrease in the influence of magnets on microbes in vitro was also detected. Conclusions : Magnets reduce the number of microbes and might be recommended as a supplement in therapy for reduced periodontal tissues. This is important because periodontal tissues that are in good conditions provide prolonged support to the oral tissues under partial and supradental denture.

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During healing following tooth extraction, inflammation and the immune response within the extraction socket are related to bone resorption. Objective : We sought to identify how the alloplastic material used for socket preservation affects the immune responses and osteoclastic activity within extraction sockets. Material and Methods : Using a porcine model, we extracted teeth and grafted biphasic calcium phosphate into the extraction sockets. We then performed a peptide analysis with samples of gingival tissue from adjacent to the sockets and compared the extraction only (EO) and extraction with socket preservation (SP) groups. We also used real-time polymerase chain reaction (PCR) to evaluate the expression level of immunoglobulins, chemokines and other factors related to osteoclastogenesis. Differences between the groups were analyzed for statistical significance using paired t tests. Results : Levels of IgM, IgG and IGL expression were higher in the EO group than in the SP group 1 week post-extraction, as were the levels of CCL3, CCL5, CXCL2, IFN-γ and TNF-α expression (p<0.05). In addition, receptor activator of nuclear factor kappa-B ligand (RANKL) was also significantly upregulated in the EO group (p<0.05), as were IL-1β, IL-6 and IL-8 (p<0.05). Conclusions : These results suggest that the beneficial effect of socket preservation can be explained by suppression of immune responses and inflammation.

Resumo em Inglês:

Recently, digital wax-up is proposed as a tool to aid prosthodontic planning. However, there are no data about the effect of prosthodontic planning on lateral occlusion scheme. Objective : This study aims to evaluate the impact of conventional and digital prosthodontic planning on lateral occlusion scheme. Material and Methods : Dental models of 10 patients were collected. All models had Angle Class I occlusion and were undergoing prosthodontic treatment that would influence the lateral occlusion scheme. Each set of models had received both conventional wax-up and digital wax-up. In relation to the lateral occlusion scheme, the following variables were evaluated: the prevalence of the different lateral occlusion scheme, number of contacting teeth and percentage of each contacting tooth. Four excursive positions on the working side were included: 0.5, 1.0, 2.0 and 3.0 mm from the maximal intercuspation position. Results : The lateral occlusion scheme of the two wax-up models was subjected to alterations following excursion. There was a tendency for the prevalence of canine-guided occlusion to increase and for the prevalence of group function occlusion to decrease with increasing excursion. The number of contacting teeth was decreasing with the increasing magnitude of excursion. For the 0.5 mm and 1.0 mm positions, the two wax-ups had significantly greater contacts than the pre-treatment models, while at the 2.0 mm and 3.0 mm positions, all the models were similar. For all models, canines were the most commonly contacting teeth, followed by the teeth adjacent to them. No difference was observed between the two wax-ups in relation to the number of contacting teeth. Conclusion : Although the prosthodontic planning had influenced the pattern of the lateral occlusion scheme and contacts, there was no difference between the conventional and digital prosthodontic planning.

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Objective : To investigate the influence of a three-dimensional cell culture model on the expression of osteoblastic phenotype in human periodontal ligament fibroblast (hPDLF) cultures. Material and Methods : hPDLF were seeded on bi-dimensional (2D) and three-dimensional (3D) collagen type I (experimental groups) and and on a plastic coverslip (control) for up to 14 days. Cell viability and alkaline phosphatase (ALP) activity were performed. Also, cell morphology and immunolabeling for alkaline phosphatase (ALP) and osteopontin (OPN) were assessed by epifluorescence and confocal microscopy. The expression of osteogenic markers, including alkaline phosphatase, osteopontin, osteocalcin (OC), collagen I (COL I) and runt-related transcription factor 2 (RUNX2), were analyzed using real-time polymerase chain reaction (RT-PCR). Mineralized bone-like nodule formation was visualized by microscopy and calcium content was assessed quantitatively by alizarin red assay. Results : Experimental cultures produced an increase in cell proliferation. Immunolabeling for OPN and ALP in hPDLF were increased and ALP activity was inhibited by three-dimensional conditions. OPN and RUNX2 gene expression was significantly higher on 3D culture when compared with control surface. Moreover, ALP and COL I gene expression were significantly higher in three-dimensional collagen than in 2D cultures at 7 days. However, at 14 days, 3D cultures exhibited ALP and COL I gene expression significantly lower than the control, and the COL I gene expression was also significantly lower in 3D than in 2D cultures. Significant calcium mineralization was detected and quantified by alizarin red assay, and calcified nodule formation was not affected by tridimensionality. Conclusion : This study suggests that the 3D cultures are able to support hPDLF proliferation and favor the differentiation and mineralized matrix formation, which may be a potential periodontal regenerative therapy.

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Injectable bone substitutes and techniques have been developed for use in minimally invasive procedures for bone augmentation. Objective : To develop a novel injectable thermo-sensitive alginate hydrogel (TSAH) as a scaffold to induce bone regeneration, using a minimally invasive tunnelling technique. Material and Methods : An injectable TSAH was prepared from a copolymer solution of 8.0 wt% Poly(N-isopropylacrylamide) (PNIPAAm) and 8.0 wt% AAlg-g-PNIPAAm. In vitro properties of the material, such as its microstructure and the sustained release of recombinant human bone morphogenetic protein-2 (rhBMP-2), were investigated. Then, with the subperiosteal tunnelling technique, this material, carrying rhBMP-2, was injected under the labial periosteum of the maxillary anterior alveolar ridge in a rabbit model. New bone formation was evaluated by means of X-ray, micro-computed tomography (micro-CT), fluorescence labelling, histological study, and immunohistochemistry study. Results : The material exhibited good injectability and thermo-irreversible properties. SEM showed an interconnected porous microstructure of the TSAH. The result of ALP activity indicated sustained delivery of BMP-2 from the TSAH from days 3 to 15. In a rabbit model, both TSAH and TSAH/rhBMP-2 induced alveolar ridge augmentation. The percentage of mineralised tissue in the TSAH/rhBMP-2 group (41.6±3.79%) was significantly higher than in the TSAH group (31.3±7.21%; p<0.05). The density of the regenerating tissue was higher in the TSAH/rhBMP-2 group than in the other groups (TSAH group, positive control, blank control; p<0.05). Conclusions : The TSAH provided convenient handling properties for clinical application. To some extent, TSAH could induce ridge augmentation and mineral deposition, which can be enhanced when combined with rhBMP-2 for a minimally invasive tunnelling injection.

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A 43-year-old woman with a unilateral cleft lip and palate, presenting a totally edentulous maxilla and mandible with marked maxillomandibular discrepancy, attended the Prosthodontics section of the Hospital for Rehabilitation of Craniofacial Anomalies, University of São Paulo for treatment. She could not close her mouth and was dissatisfied with her complete dentures. Treatment planning comprised placement of six implants in the maxilla, four in the mandible followed by prostheses installation and orthognathic surgery. The mandibular full arch prosthesis guided the occlusion for orthognathic positioning of the maxilla. The maxillary complete prosthesis was designed to assist the orthognathic surgery with a provisional prosthesis (no metal framework), allowing reverse treatment planning. Maxillary and mandibular realignment was performed. Three months later, a relapse in the position of the maxilla was observed, which was offset with a new maxillary prosthesis. This isa complex interdisciplinary treatment and two-year follow-up is presented and discussed. It should be considered that this type of treatment could also be applied in non-cleft patients.