Histologic Study of Urethral Extracellular Matrix and Collagen From Aging and Long-Term Alloxan-induced Diabetic Male Rats

OBJECTIVES To evaluate the histological alterations of extracellular matrix in long-term alloxan-induced diabetes and aging urethras of male rats with descriptions of total connective tissue, muscle layer and collagen types I and III relative amounts. METHODS Histologic evaluations were performed in 3 animal groups: group 1, 8 weeks old; group 2, 44 weeks old; and group 3, 44 weeks old with alloxan-induced diabetes. The muscle layer thickness, extracellular matrix fibrosis, and collagen were quantified on digital images of the urethral samples. RESULTS A higher total thickness and muscle layer thickness and higher connective tissue and collagen content were observed in the urethras of group 3. No changes in the collagen type III/I ratio were found in the urethra of groups 2 and 3. CONCLUSIONS Our results suggest that the morphologic alterations of the urethra should also be considered in long-term studies of diabetic lower urinary tract dysfunction. These morphologic alterations due to diabetes differ from the changes induced by aging itself and could represent a final stage in decompensate urethras. Further studies are necessary to establish the real influence of the urethral morphologic changes on lower urinary tract diabetes dysfunction. UROLOGY 77: 510.e6-510.e11, 2011. (C) 2011 Elsevier Inc.

Hypertrophy and neuron loss: structural changes in sheep SCG induced by unilateral sympathectomy

Recently, superior cervical ganglionectomy has been performed to investigate a variety of scientific topics from regulation of intraocular pressure to suppression of lingual tumour growth. Despite these recent advances in our understanding of the functional mechanisms underlying superior cervical ganglion (SCG) growth and development after surgical ablation, there still exists a need for information concerning the quantitative nature of the relationships between the removed SCG and its remaining contralateral ganglion and between the remaining SCG and its modified innervation territory. To this end, using design-based stereological methods, we have investigated the structural changes induced by unilateral ganglionectomy in sheep at three distinct timepoints (2, 7 and 12 weeks) after surgery. The effects of time, and lateral (left-right) differences, were examined by two-way analyses of variance and paired t-tests. Following removal of the left SCG, the main findings were: (i) the remaining right SCG was bigger at shorter survival times, i.e. 74% at 2 weeks, 55% at 7 weeks and no increase by 12 weeks, (ii) by 7 weeks after surgery, the right SCG contained fewer neurons (no decrease at 2 weeks, 6% fewer by 7 weeks and 17% fewer by 12 weeks) and (iii) by 7 weeks, right SCG neurons were also larger and the magnitude of this increase grew substantially with time (no rise at 2 weeks, 77% by 7 weeks and 215% by 12 weeks). Interaction effects between time and ganglionectomy-induced changes were significant for SCG volume and mean perikaryal volume. These findings show that unilateral superior cervical ganglionectomy has profound effects on the contralateral ganglion. For future investigations, it would be interesting to examine the interaction between SCGs and their innervation targets after ganglionectomy. Is the ganglionectomy-induced imbalance between the sizes of innervation territories the milieu in which morphoquantitative changes, particularly changes in perikaryal volume and neuron number, occur? Mechanistically, how would those changes arise? Are there any grounds for believing in a ganglionectomy-triggered SCG cross-innervation and neuroplasticity? (C) 2011 ISDN. Published by Elsevier Ltd. All rights reserved.

Influence of Final Rinse Technique on Ability of Ethylenediaminetetraacetic Acid of Removing Smear Layer

Introduction: There is ongoing debate regarding the ideal sequence, volume, and concentration of irrigants, length of time for irrigation, and irrigation technique to achieve debridement of the root canal system. The aim of this study was to verify the impact of the final rinse technique on smear layer removal ability of 17% ethylenediaminetetraacetic acid (EDTA). Methods: Sixteen single-rooted human teeth were instrumented and divided into 2 groups at the final rinse step according to the following final rinse techniques used: continuous rinse group, continuous rinse with EDTA during 3 minutes, and rinse and soaking group, rinse with 1 mL of EDTA, soaking of the canal for 2 minutes and 30 seconds, and rinse completion with the remaining 4 mL for 30 seconds. The specimens were split lengthwise and observed under scanning electron microscope. Results: Data were analyzed with Kruskal-Wallis and Dunn tests. The continuous rinse group presented more debris-free surfaces when compared with the rinse and soaking group (P <. 01). When the root canal areas were compared within the groups, no statistical differences were found (P > .05). Conclusions: It can be concluded that a continuous rinse with 5 mL of EDTA for 3 minutes can more efficiently remove the smear layer from root canal walls. (J Endod 2010;36:512-514)

Influence of Blood Contamination on Bond Strength of a Self-etching Adhesive to Dental Tissues

Purpose: The aim of this study was to detect the influence of (1) storage period of heparinized blood, (2) type of blood and presence of contaminant, (3) application mode of cleansing agents, and (4) efficacy of cleansing agents on contaminated enamel and dentin during the adhesion process of a one-step adhesive system. Materials and Methods: One hundred four human molars were sectioned into halves along the long axis for enamel and dentin tests. Heparinized and fresh blood were obtained from the same donor, applied and dried to maintain a layer of dry blood on the top of samples. The cleansing agents used were hydrogen peroxide, anionic detergent, and antiseptic solution. A one-step adhesive system (Clearfil S3 Bond) was applied on the dental surface, and composite resin cylinders were built up using Tygon tubing molds. After 24 h, the mu SBS test (1 mm/min) and fracture analysis were performed. Results: There was no statistically significant difference in bond strength values regarding the storage period of heparinized blood and the types of blood. Groups without contamination presented higher bond strengths than contaminated groups. The application mode of the cleansing agents had no influence on bond strength results. There was no statistically significant difference among cleansing agents and they were as effective as a water stream in counteracting the effect of blood contamination. Conclusion: Heparinized blood can be used as a contaminant for up to one week, and it is a reliable procedure to standardize the contaminant. The cleansing agents can be used without friction. A water stream is sufficient to remove blood contamination from dental tissues, before the application of a one-step adhesive system.

Influence of different volumes of EDTA for final rinse on smear layer removal

Objective. The aim of this study was to analyze the influence of different volumes of 17% EDTA for final rinse on smear layer removal on the different areas of the root canal. Study design. Forty single-rooted teeth were instrumented using rotary instruments. The teeth were divided into 3 test groups according to the EDTA volume for final rinse (5 mL, 10 mL, 15 mL) and 1 control group (10 mL of 1% sodium hypochlorite). The roots were axially split into halves, and the smear layer removal from the canals was determined under scanning electron microscope. The data were analyzed using Kruskal Wallis and Dunn tests. Results. The 3 experimental groups showed no statistical difference (P > .05); however, when the test groups were compared to the control group statistically significant differences were found (P < .01). The root canal wall surfaces of teeth in the control group showed the presence of heavy smear layer through the entire length of the root canals. The other groups showed mainly smear layer-free surfaces or a small amount of debris. When coronal, middle and apical thirds were compared, there was no statistically significant difference (P > .05). Conclusions. Based on the results, it appears that a final rinse with 5 mL of EDTA per canal provides good smear layer removal, with root canal walls free of debris and mostly open dentinal tubules in all areas. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2008;106:e40-e43)

GAPD and tubulin are suitable internal controls for qPCR analysis of oral squamous cell carcinoma cell lines

The selection of housekeeping genes is critical for gene expression studies. To address this issue, four candidate housekeeping genes, including several commonly used ones, were investigated in oral squamous cell carcinoma cell lines. A simple quantitative RT-PCR approach was employed by comparing relative expression of the four candidate genes within two cancerous cell tines (HN6 and HN31) and one noncancerous cell tine (HaCaT) treated or not with EGF and TGF-beta 1. Data were analyzed using ANOVA followed by the NormFinder software program. On this basis, stability of the candidate housekeeping genes was ranked and non statistical differences were found using ANOVA test. On the other hand, the NormFinder was able to show that GAPD and TUBB presented the less variable results, representing appropriated housekeeping genes for the samples and conditions anatyzed. In conclusion, this study suggests that the GAPD and the TUBB represent adequate normalizers for gene profiting studies in OSCC cell. tines, covering, respectively, high and low expression levels genes. (C) 2008 Elsevier Ltd. All rights reserved.

Effect of the Combination of Sodium Hypochlorite and Chlorhexidine on Dentinal Permeability and Scanning Electron Microscopy Precipitate Observation

Introduction: This study compared the combined use of sodium hypochlorite (NaOCl) and chlorhexidine (CXH) with citric acid and CXH on dentinal permeability and precipitate formation. Methods: Thirty-four upper anterior teeth were prepared by rotary instrumentation and NaOCl. The root canal surfaces were conditioned for smear layer removal using 15% citric acid solution under ultrasonic activation and a final wash with distilled water. All teeth were dried, and 30 specimens were randomly divided into three equal groups as follows: positive control group (PC), no irrigation; 15% citric acid + 2% CHX group (CA + CHX); and 1% NaOCl + 2% CHX group (NaOCl + CHX). All roots were immersed in a 0.2% Rhodamine B solution for 24 hours. One-millimeter-thick slices from the cementum-enamel junction were scanned at 400 dpi and analyzed using the software ImageLab (LIDO-USP, Sao Paulo, Brazil) for the assessment of leakage in percentage. For scanning electron microscopy analysis, four teeth, irrigated for NaOCl + CHX samples, were split in half, and each third was evaluated at 1,000x and 5,000x (at the precipitate). Results: Using the analysis of variance test followed by the Bonferroni comparison method, no statistical differences between groups were found when analyzed at the cervical and medium thirds. At the apical third, differences between the PC and NaOCl + CHX (p<0.05) and CA + CHX and NaOCl + CHX could be seen (p < 0.05). Conclusion: The combination of 1% NaOCl and 2% CHX solutions results in the formation of a flocculate precipitate that acts as a chemical smear layer reducing the dentinal permeability in the apical third. (J Endod 2010;36:847-850)

Compromised Bond Strength after Root Dentin Deproteinization Reversed with Ascorbic Acid

Introduction: The present study evaluated the effect of a reducing agent on the bond strength of deproteinized root canal dentin surfaces when using a self-adhesive versus dual-cured cement. Regional differences were also evaluated. Methods: A total of 45 bovine incisor roots were divided into 3 groups: irrigation with physiologic solution (control), 10-minute deproteinization with 5% NaOCl, and 10-minute deproteinization with 5% NaOCl followed by 10 minutes of 10% ascorbic acid. Fiber posts were cemented with either RelyX 0100 or RelyX ARC (with SingleBond 2 or Clearfil SE Bond). The push-out bond strength was evaluated after 24 hours of storage. Data were submitted to three-way analyses of variance and Dunnett 13 tests (alpha = 0.05). Results: No differences between cements were observed within the testing conditions, regardless of the adhesive (P < .05). Deproteinization reduced bond strengths. Subsequent treatment with ascorbic acid was capable of reversing bond strength value changes to levels similar to those of controls. Regional radicular differences were also found, where coronal > middle > apical. Conclusions: The reducing agent was capable. of reversing the effect of dentin deproteinization, and RelyX U100 behaved similarly to RelyX ARC. (J Endod 2010;36:130-134)

Histological evaluation of the effectiveness of increased apical enlargement for cleaning the apical third of curved canals

P>Objective To evaluate the influence of apical size on cleaning of the apical third of curved canals prepared with rotary instruments. Methodology Forty-four mesiobuccal canals of maxillary molars teeth were instrumented to different apical sizes (30, 0.02; 35, 0.02; 40, 0.02; 45, 0.02) using a crown-down technique. After canal preparation, the apical thirds of the roots were submitted to histological processing and examination. The specimens were analysed at 40x magnification and the images were submitted to morphometric analysis with an integration grid to evaluate the percentage of debris and uninstrumented root canal walls. The action of the instruments on the root canal walls was assessed based on the surface regularity, abrupt change on the continuity of root canal walls, and partial or total pre-dentine removal. The results were statistically compared using one-way anova with post hoc Tukey test. Pearson`s correlation was performed to identify potential correlations between values. Results The percentage of uninstrumented root canal dentine was higher when apical enlargement was performed with instruments 30, 0.02 taper (55.64 +/- 4.62%) and 35, 0.02 taper (49.03 +/- 5.70%) than with instruments 40, 0.02 taper (38.08 +/- 10.44%) and 45, 0.02 taper (32.65 +/- 8.51%) (P < 0.05). More debris were observed when apical enlargement was performed with instruments 30, 0.02 taper (34.62 +/- 9.49%) and 35, 0.02 taper (25.33 +/- 7.37%) (P < 0.05). There was a significant correlation between the amount of remaining debris and the perimeter of uninstrumented root canal dentine (r = 0.9130, P < 0.001). Conclusion No apical enlargement size allowed the root canal walls to be prepared completely. Apical third cleanliness could be predicted by instrument diameter.

Determination of para-Chloroaniline and Reactive Oxygen Species in Chlorhexidine and Chlorhexidine Associated with Calcium Hydroxide

The aim of this study was to determine whether para-chloroaniline (PCA) and/or reactive oxygen species (ROS) are generated by chlorhexidine (CHX) alone or after CHX is mixed with calcium hydroxide at different time points. Mass spectrometry was performed to detect PCA in samples of 0.2% CHX and Ca(OH)2 mixed with 0.2% CHX. High-performance liquid chromatography was used to confirm the presence of CHX in the mixture with Ca(OH)2. The samples were analyzed immediately after mixing and after 7 and 14 days. During the intervals of the experiment, the samples were maintained at 36.5 degrees C and 95% relative humidity. PCA was detected in the 0.2% CHX solution after 14 days. The mixture of CHX with Ca(CH)2 liberated ROS at all time points, but no traces of CHX were present in the mixture as a result of immediate degradation of the CHX. (J Endod 2008;34:1508-1514)

Effect of 980-nanometer diode laser on root canal permeability after dentin treatment with different chemical solutions

This study evaluated the effect of 980-nm diode laser at different parameters on root canal dentin permeability associated with different irrigants. Seventy-five canines were sectioned at 15 mm from the apex, prepared mechanically up to #40 .02 instrument, and irrigated with 2 mL distilled water. Final irrigation (10 mL) was used as follows: (1) distilled water; (2) 1% NaOCl; (3) 17% ethylenediaminetetraacetic acid + a cationic surfactant cetyltrimethylammonium bromide (EDTAC). Laser was applied at 1.5 or 3.0 W as either continuous wave or pulsed wave (100 Hz). The teeth were then processed histochemically, the percentage of copper ion penetration into the dentin of the canal walls was counted, and the data were analyzed statistically with the Tukey-Kramer test (alpha < .01). When laser was associated with water, an increase in permeability was found, whereas permeability decreased when associated with EDTAC. Dentin permeability after laser irradiation was directly dependent on the solution used for final irrigation.

Effect of Chelating Solutions on the Microhardness of Root Canal Lumen Dentin

Introduction: The greatest reduction in microhardness of the most superficial layer of dentin of the root canal lumen is desired. The use of chelating agents during biomechanical preparation of root canals removes smear layer, increasing the access of the irrigant into the dentin tubules to allow adequate disinfection, and also reduces dentin microhardness, facilitating the action of endodontic instruments. This study evaluated the effect of different chelating solutions on the microhardness of the most superficial dentin layer from the root canal lumen. Methods: Thirty-five recently extracted single-rooted maxillary central incisors were instrumented, and the roots were longitudinally sectioned in a mesiodistal direction to expose the entire canal extension. The specimens were distributed in seven groups according to the final irrigation: 15% EDTA, 10% citric acid, 5% malic acid, 5% acetic acid, apple vinegar, 10% sodium citrate, and control (no irrigation). A standardized volume of 50 mu L of each chelating solution was used for 5 minutes. Dentin microhardness was measured with a Knoop indenter under a 10-g load and a 15-second dwell time. Data were analyzed statistically by one-way analysis of variance and Tukey-Kramer multiple-comparison test at 5% significance level. Results: EDTA and citric acid had the greatest overall effect, causing a sharp decrease in dentin microhardness without a significant difference (p > .05) from each other. However, both chelators differed significantly from the other solutions (p < .001). Sodium citrate and deionized water were similar to each other (p > .05) and did not affect dentin microhardness. Apple vinegar, acetic acid, and malic acid were similar to each other (p > .05) and presented intermediate results. Conclusion: Except for sodium citrate, all tested chelating solutions reduced microhardness of the most superficial root canal dentin layer. EDTA and citric acid were the most efficient. (J Endod 2011;37:358-362)