Proteomic analysis of kidney in rats chronically exposed to fluoride

Two-dimensional gel electrophoresis (2-DE) was used to better understand alterations in renal metabolism induced by fluoride (F). Three groups of weanling male Wistar rats were treated with drinking water containing 0 (control), 5, or 50 ppm F for 60 days (n=6/group). Kidneys were collected for proteomic and histological (HE) analysis. After protein isolation, renal proteome profiles were examined using 2-DE and Colloidal Coomassie Blue staining. Protein spots with a 2-fold significant difference as detected by quantitative intensity analysis (image Master Platinum software) and t-test (p < 0.05) were excised and analyzed by MALDI-TOF MS (matrix assisted laser desorption ionization-time-of-flight mass spectrometry). The histological analysis revealed no damage in kidneys induced by F, except for a vascular congestion in the 50 ppm F group. Between control vs 50 ppm F, and control vs 5 ppm F groups, 12 and 6 differentially expressed proteins were detected, respectively. Six proteins, mainly related with metabolism, detoxification and housekeeping, were successfully identified. At the high F group, pyruvate carboxylase, a protein involved in the formation of oxaloacetate was found to be downregulated, while enoyl coenzyme A hydratase, involved in fatty acids oxidation, was found to be upregulated. Thus, proteomic analysis can provide new insights into the alterations in renal metabolism after F exposure, even in low doses. (C) 2009 Elsevier Ireland Ltd. All rights reserved.

Bone repair and augmentation using block of sintered bovine-derived anorganic bone graft in cranial bone defect model

To histomorphometrically investigate the repair of critical size defects (CSDs) and bone augmentation in cranial walls using block of sintered bovine-derived anorganic bone (sBDAB) graft. Forty guinea-pigs were divided into test (n=20) and CSD control (n=20) groups. In each animal, a full-thickness bone defect with 9.5 mm diameter was made in the frontal bone. The defects were filled with an sBDAB block soaked in blood in the test group and with blood clot in the CSD control group. The skulls were collected at 0 h (n=2) and 30, 90 and 180 days (n=6/group and period) postoperatively. The volume density and total volume of newly formed bone, sBDAB, blood vessels and connective tissue, vertical thickness of removed bone plug, sBDAB block and graft area were evaluated. The vertical thickness of the adapted sBDAB block was 3.8 times higher than that of the removed bone plug and did not show significant difference between periods, filling in average 29.8% of the total graft region. The sBDAB block exhibited complete osseointegration with the borders of the defect at 90 days. At 90 and 180 days, the vertical thickness of the graft was 279% in the average, and the total volume of bone augmentation was, respectively, 78.8% and 148.5% higher compared with the removed bone plug. The defects of the CDS control group showed limited osteogenesis and filling by connective tissue plus tegument. The sBDAB block can be used to promote repair of CSDs and bone augmentation in the craniomaxillofacial region, due to its good osteoconductive and slow resorptive properties. To cite this article:Cestari TM, Granjeiro JM, de Assis GF, Garlet GP, Taga R. Bone repair and augmentation using block of sintered bovine-derived anorganic bone graft in cranial bone defect model.Clin. Oral Impl. Res. 20, 2009; 340-350.doi: 10.1111/j.1600-0501.2008.01659.x.

The role of surface sealants in the roughness of composites after a simulated toothbrushing test

Objectives: To evaluate the influence of two surface sealants (BisCover/Single Bond) and three application techniques (unsealed/conventional/co-polymerization) on the roughness of two composites (Filtek Z250/Z350) after the toothbrushing test. Methods: Seventy-two rectangular specimens (5 mm x 10 mm x 3 mm) were fabricated and assigned into 12 groups (n = 6). Each sample was subjected to three random roughness readings at baseline, after 100,000 (intermediate), and 200,000 (final) toothbrushing strokes. Roughness (R) at each stage was obtained by the arithmetic mean of the reading of each specimen. Sealant removal was qualitatively examined (optical microscope) and classified into scores (0-3). Data were analyzed by Student`s paired t-test, two-way ANOVA/Tukey`s test, and by Wilcoxon, Kruskal-Wallis and Miller`s test (alpha = 0.05). Results: Z250 groups at baseline did not differ statistically from each other. Unsealed Z350 at baseline had lower R values. All the unsealed groups presented gradual decrease in R from baseline to final brushing. From baseline to the inter-mediate stage, Z250 co-polymerized groups presented a significant reduction in R (score 3). Conventionally sealed groups had no significant changes in R (scores 2-0.8). From baseline to the intermediate stage, the conventionally sealed Z350 Single Bond group had an increase in R (score 1.5). In the final stage, all the conventionally sealed groups presented a reduction in R (scores 0.7-0). Co-polymerized Single Bond groups had a significant reduction in R (scores 2.5-2.7), and co-polymerized BisCover groups an increase in R (scores 2.8-3). Conclusions: At any brushing stage, sealed composites presented superior performance when compared with unsealed composites. (C) 2009 Elsevier Ltd. All rights reserved.

Evaluation of the temperature of different refrigerant sprays used as a pulpal test

The temperature of different refrigerant sprays (Endo-Ice, Endo-Frost, Coolermatic and Sprayon Contact and Tuner Cleaner) used as pulpal tests were evaluated in vitro. A thermocouple placed inside the pulp chamber of a maxillary central incisor was used to register the temperature changes when the refrigerant sprays were applied with a cotton swab, for 10 s. Results indicate that Endo-Ice and Endo-Frost presented the lowest temperatures among the refrigerant sprays tested. Temperatures measured inside the pulp chamber, however, were statistically similar in all groups.

Quantitative analysis of Langerhans` cells in oral chronic graft-vs.-host disease

The Langerhans cells (LCs) are scattered throughout the epithelium of skin and mucosa and have been associated with the graft-vs.-host disease (GVHD), which is the highest cause of morbidity and mortality in patients who underwent bone marrow transplant (BMT). This study aims at quantifying the LCs in the oral chronic GVHD (cGVHD). Microscopic sections from biopsies carried out in the buccal mucosa of 40 patients who underwent allogenic BMT and developed (20) or not (20) oral cGVHD (Groups 1 and 2, respectively) were utilised. For the control group, free surgical margins of 20 biopsies of non-inflammatory lesions in the buccal mucosa (Group 3) were used. The sections were studied in routine colouration and immunostained for CD1a. Group 1 (with cGVHD) presented a greater number of Langerhans` cells/mm(2) (50.6 +/- 37.2) when compared with the other groups (Group 2, 23.11 +/- 19.7; Group 3, 16.6 +/- 17.3). Our results suggest a greater recruitment of LCs in patients transplanted with cGVHD, probably as a result of cytokines secreted by the inflammatory cells.

Inflammatory Papillary Hyperplasia of the Palate: Quantitative Analysis of Candida albicans and its Negative Correlation with Microscopic and Demographic Aspects

Inflammatory papillary hyperplasia of the palate (IPHP) is a tissue-reactive overgrowth characterized by hyperemic mucosa with nodular or papillary appearance in the palate. The exact pathogenesis is still unclear. In this study, the presence of Candida albicans in the epithelial lining was evaluated using the indirect immunofluorescence staining technique. Strongly stained C albicans was observed only in the lesions of the IPHP group. Therefore, the detection of C albicans in almost all samples from IPHP tissue enabled a suggestion as to the microbial etiology of the disease, since the use of dental prostheses was reported. Int J Prosthodont 2011;24:235-237

Progressive recovery of osseoperception as a function of the combination of implant-supported prostheses

The extraction of teeth involves the elimination of extremely sensitive periodontal mechanoreceptors, which play an important role in oral sensory perception. Objectives: The aim of this study was to evaluate the recovery of interocclusal sensory perception for micro-thickness in individuals with different types of implant-supported prostheses. Materials and Methods: Wearers of complete dentures (CDs) comprised the negative control group (group A, n=17). The experimental group consisted of wearers of prostheses supported by osseointegrated implants (Group B, n=29), which was subsequently divided into 4 subgroups: B(1) (n=5) - implant supported overdentures (ISO) occluding with CD; B(2) (n=6) - implant-supported fixed prostheses (ISFP) occluding with CD; B3 (n=8) - wearers of maxillary and mandibular ISFP, and B(4) (n=10) - ISFP occluding with natural dentition (ND). Individuals with ND represented the positive control group (Group C, n=24). Aluminum foils measuring 10 mu m, 24 mu m, 30 mu m, 50 mu m, 80 mu m, and 104 mu m thickness were placed within the premolar area, adding up to 120 tests for each individual. Results: The mean tactile thresholds of groups A, B(1), B(2), B(3), B(4), and C were 92 mu m, 27 mu m, 27 mu m, 14 mu m, 10 mu m, and 10 mu m, respectively. [Correction added after publication online 18 April 2008: in the preceding sentence 92 mu m, 27 mu m, 14 mu m, 10 mu m and 10 mu m, was corrected to 92 mu m, 27 mu m, 27 mu m, 14 mu m, 10 mu m and 10 mu m.] The Kruskal-Wallis test revealed significant difference among groups (P < 0.05). The Dunn test revealed that group A was statistically different from groups C, B(3), and B(4), and that B(1) and B(2) were statistically different from group C. Conclusion: Progressive recovery of osseoperception as a function of the combination of implant-supported prostheses could be observed. Moreover, ISO and/or ISFP combinations may similarly maximize the recovery of osseoperception.

Analysis of enamel microbiopsies in shed primary teeth by Scanning Electron Microscopy (SEM) and Polarizing Microscopy (PM)

The aims of this study were 1) to verify how close to the theoretically presumed areas are the areas of enamel microbiopsies carried out in vivo or in exfoliated teeth; 2) to test whether the etching solution penetrates beyond the tape borders: 3) to test whether the etching solution demineralizes the enamel in depth. 24 shed upper primary central incisors were randomly divided into two groups: the Rehydrated Teeth Group and the Dry Teeth Group. An enamel microbiopsy was performed, and the enamel microbiopsies were then analyzed by Scanning Electron Microscopy (SEMI) and Polarizing Microscopy (PM). Quantitative birefringence measurements were performed. The ""true"" etched area was determined by measuring the etched enamel using the NIH Image analysis program. Enamel birefringence was compared using the paired t test. There was a statistically significant difference when the etched areas in the Rehydrated teeth were compared with those of the Dry teeth (p = 0.04). The etched areas varied from -11.6% to 73.5% of the presumed area in the Rehydrated teeth, and from 6.6% to 61.3% in the Dry teeth. The mean percentage of variation in each group could be used as a correction factor for the etched area. Analysis of PM pictures shows no evidence of in-depth enamel demineralization by the etching solution. No statistically significant differences in enamel birefringence were observed between values underneath and outside the microbiopsy area in the same tooth, showing that no mineral loss occurred below the enamel superficial layer. Our data showed no evidence of in-depth enamel demineralization by the etching solution used in the enamel microbiopsy proposed for primary enamel. This study also showed a variation in the measured diameter of the enamel microbiopsy in nineteen teeth out of twenty four, indicating that in most cases the etching solution penetrated beyond the tape borders. (C) 2009 Elsevier B.V. All rights reserved.

A simple, inexpensive and easily reproducible model of spinal cord injury in mice: Morphological and functional assessment

Spinal cord injury (SCI) causes motor and sensory deficits that impair functional performance, and significantly impacts life expectancy and quality. Animal models provide a good opportunity to test therapeutic strategies in vivo. C57BL/6 mice were subjected to laminectomy at T9 and compression with a vascular clip (30 g force, 1 min). Two groups were analyzed: injured group (SCI, n = 33) and laminectomy only (Sham, n = 15). Locomotor behavior (Basso mouse scale-BMS and global mobility) was assessed weekly. Morphological analyses were performed by LM and EM. The Sham group did not show any morphofunctional alteration. All SCI animals showed flaccid paralysis 24 h after injury. with subsequent improvement. The BMS score of the SCI group improved until the intermediate phase (2.037 +/- 1.198): the Sham animals maintained the highest BMS score (8.981 +/- 0.056). p < 0.001 during the entire time. The locomotor speed was slower in the SCI animals (5.581 +/- 0.871) than in the Sham animals (15.80 +/- 1.166), p < 0.001. Morphological analysis of the SCI group showed, in the acute phase, edema, hemorrhage, multiple cavities, fiber degeneration, cell death and demyelination. In the chronic phase we observed glial scarring, neuron death, and remyelination of spared axons by oligodendrocytes and Schwann cells. In conclusion, we established a simple, reliable, and inexpensive clip compression model in mice, with functional and morphological reproducibility and good validity. The availability of producing reliable injuries with appropriate outcome measures represents great potential for studies involving cellular mechanisms of primary injury and repair after traumatic SCI. (C) 2008 Elsevier B.V. All rights reserved.

Photoelastic Study of the Support Structures of Distal-Extension Removable Partial Dentures

Purpose: The double system of support, in which the distal-extension removable partial denture adapts, causes inadequate stress around abutment teeth, increasing the possibility of unequal bone resorption. Several ways to reduce or more adequately distribute the stress between abutment teeth and residual ridges have been reported; however, there are no definitive answers to the problem. The purpose of this study was to analyze, by means of photoelasticity, the most favorable stress distribution using three retainers: T bar, rest, proximal plate, I bar (RPI), and circumferential with mesialized rest. Materials and Methods: Three photoelastic models were made simulating a Kennedy Class II inferior arch. Fifteen dentures with long saddles, five of each design, were adjusted to the photoelastic patterns and submitted first to uniformly distributed load, and then to a load localized on the last artificial tooth. The saddles were then shortened and the tests repeated. The quantitative and qualitative analyses of stress intensity were done manually and by photography, respectively. For intragroup analyses the Wilcoxon test for paired samples was used, while for intergroup analyses Friedman and Wilcoxon tests were used to better identify the differences (p < 0.05). Results: The RPI retainer, followed by the T bar, demonstrated the best distribution of load between teeth and residual ridge. The circumferential retainer caused greater concentration of stress between dental apexes. Stress distribution was influenced by the type of retainer, the length of the saddle, and the manner of load application. Conclusions: The long saddles and the uniformly distributed loads demonstrated better distribution of stress on support structures.

Bond Strength of Fiber Posts to Weakened Roots After Resin Restoration With Different Light-Curing Times

Introduction: This study evaluated the bond strength of translucent fiber posts to experimentally weakened radicular dentin restored with composite resin and polymerized with different light-exposure time. Methods: Roots of 60 maxillary incisors were used. Twenty-four hours after obturation, the filling materials of root canals were removed to a depth of 12 mm, and 4 groups were randomly formed. In 3 groups, root dentin was flared to produce a space between fiber post and canal walls. In the control group, the roots were not experimentally weakened. The flared roots were bulk restored with composite resin, which was light-activated through the translucent post for 40, 80, or 120 seconds. Posts were cemented, and after 24 hours, all roots were sectioned transversely in the coronal, middle, and apical regions, producing 1-mm-thick slices. Push-out test was performed, and failure modes were observed. Results The quantitative analysis showed significant statistical difference only among groups (P <.001). Comparing the weakened/restored groups, composite light-exposure time did not influence the results. Overall, adhesive failures occurred more frequently than other types of failures. Cohesive failures occurred only in the weakened/restored roots. Conclusions Intracanal root restoration with composite resin and translucent fiber posts provided similar or higher bond strength to dentin than the control group, regardless of the light-exposure time used for polymerization. (J Endod 2009;35:1034-1039)

Quantitative 3D profilometry and SEM analysis of the adaptation of root-end filling materials placed under an optical microscope

P>Aim To evaluate by 3D profilometry and scanning electron microscopy (SEM), the marginal adaptation of mineral trioxide aggregate (MTA) and Sealer 26 placed in root-end cavities with direct vision or under an optical microscope. Methodology The root ends of 52 root filled canine teeth were filled with MTA or Sealer 26 under direct vision or optical microscope (n = 13). In each group, eight specimens were analysed by profilometry for measurement of the area and depth of gaps. In the other five specimens, gap area was measured using SEM to verify marginal adaptation and surface characteristic. Data were analysed by parametric (anova and Tukey) and non-parametric (Kruskal-Wallis and Dunn) tests. Results The assessment of the adaptation of both materials to dentine was not influenced by the mode of visualization, which was confirmed by both profilometry and SEM observations. The voids measured with profilometry for Sealer 26 under direct vision were significantly wider and deeper than those for MTA under direct vision (P < 0.05). In SEM, significantly larger gap areas were observed with Sealer 26 (P < 0.05). Conclusion Root-end cavities filled with MTA had smaller gaps and better marginal adaptation than Sealer 26.