Evaluation of epoxy resin sealer after three root canal filling techniques by confocal laser scanning microscopy

The aim of this study was to evaluate the penetration of endodontic sealer into the dentin tubules, the integrity of the sealer layer perimeter, and the sealer area at the apical third after different filling techniques by confocal laser scanning microscopy (CLSM). Forty-five mandibular premolars were mechanically prepared with ProTaper files, until F5 file. Thereafter, they were filled with an epoxy-resin sealer (AH Plus) mixed with Rhodamine B dye (0.1% proportion) and allocated in three groups: Group 1, single master cone; Group 2, cold lateral compaction; and Group 3, Thermafil. For confocal laser scanning microscopy analysis, the specimens were transversely sectioned at 4 mm from the apex. The images at x10 and x40 were analyzed by Imagetool 3.0 software. Significant differences were not found among the three experimental groups according the dentin-impregnate area by the sealer (P = 0.68) and between the sealer and root canal perimeter (P = 0.18). However, root canal filling techniques were significantly different when apical sealer areas were compared (P = 0.001). Thermafil group showed smaller sealer areas (8.09%) while cold lateral compaction and gutta-percha master cone showed similar areas (17.37 and 21.18%, respectively). The dentin-impregnated area was not dependent on the root canal filling technique. Single master cone, cold lateral condensation and Thermafil techniques presented integrity of the sealer perimeter close to 100% and Thermafil resulted in a significantly thinner sealer layer. Microsc. Res. Tech. 75:12771280, 2012. (C) 2012 Wiley Periodicals, Inc.

Microtensile bond strength of resin composite to dentin treated with Er:YAG laser of bleached teeth

The objective of this study was to evaluate the influence of Er:YAG laser (lambda = 2.94 mu m) on microtensile bond strength (mu TBS) and superficial morphology of bovine dentin bleached with 16% carbamide peroxide. Forty bovine teeth blocks (7 x 3 x 3 mm(3)) were randomly assigned to four groups: G1- bleaching and Er:YAG irradiation with energy density of 25.56 J/cm(2) (focused mode); G2 - bleaching; G3 - no-bleaching and Er:YAG irradiation (25.56 J/cm(2)); G4 - control, non-treated. G1 and G2 were bleached with 16% carbamide peroxide for 6 h during 21 days. Afterwards, all blocks were abraded with 320 to 600-grit abrasive papers to obtain flat standardized dentin surfaces. G1 and G3 were Er:YAG irradiated. Blocks were immediately restored with 4-mm-high composite resin (Adper Single Bond 2, Z-250-3 M/ESPE). After 24 h, the restored blocks (n = 9) were serially sectioned and trimmed to an hour-glass shape of approximately 1 mm(2) at the bonded interface area, and tested in tension in a universal testing machine (1 mm/ min). Failure mode was determined at a magnification of 100x using a stereomicroscope. One block of each group was selected for scanning electron microscope (SEM) analysis. mu TBS data was analyzed by two-way ANOVA and Tukey test (alpha = 0.05). Mean bond strengths (SD) in MPa were: G1- 32.7 (5.9)(A); G2- 31.1 (6.3)(A); G3- 25.2 (8.3)(B); G4- 36.7 (9.9).(A) Groups with different uppercase letters were significantly different from each other (p < .05). Enamel bleaching procedure did not affect mu TBS values for dentin adhesion. Er:YAG laser irradiation with 25.56 J/cm(2) prior to adhesive procedure of bleached teeth did not affect mu TBS at dentin and promoted a dentin surface with no smear layer and opened dentin tubules observed under SEM. On the other hand, Er:YAG laser irradiation prior to adhesive procedure of non-bleached surface impaired mu TBS compared to the control group.

Antimicrobial effect of endodontic solutions used as final irrigants on a dentine biofilm model

Aim To evaluate the residual biovolume of live bacterial cells, the mean biofilm thickness and the substratum coverage found in mixed biofilms treated with different endodontic irrigant solutions. Methodology Twenty-five bovine dentine specimens were infected intraorally using a removable orthodontic device. Five samples were used for each irrigant solution: 2% chlorhexidine, 1% sodium hypochlorite (NaOCl), 10% citric acid, 17% EDTA and distilled water. The solutions were used for 5 min. The samples were stained using the Live/Dead technique and evaluated using a confocal microscope. Differences in the amount of total biovolume (mu m3), number of surviving cells (mu m3), mean biofilm thickness (mu m) and substratum coverage (%) of the treated biofilms were determined using nonparametric statistical tests (P < 0.05). Results Similar values of biovolume total, biovolume of live subpopulations and substratum coverage were found in 2% chlorhexidine, 10% citric acid, 17% EDTA and distilled water-treated biofilms (P > 0.05). The lower values of the studied parameters were found in 1% NaOCl-treated dentine (P < 0.05) with the exception of the mean biofilm height criteria that did not reveal significant differences amongst the irrigant solutions (P > 0.05). Conclusions One per cent sodium hypochlorite was the only irrigant that had a significant effect on biofilm viability and architecture.

Anisotropic Brownian motion in ordered phases of DNA fragments

Using Fluorescence Recovery After Photobleaching, we investigate the Brownian motion of DNA rod-like fragments in two distinct anisotropic phases with a local nematic symmetry. The height of the measurement volume ensures the averaging of the anisotropy of the in-plane diffusive motion parallel or perpendicular to the local nematic director in aligned domains. Still, as shown in using a model specifically designed to handle such a situation and predicting a non-Gaussian shape for the bleached spot as fluorescence recovery proceeds, the two distinct diffusion coefficients of the DNA particles can be retrieved from data analysis. In the first system investigated (a ternary DNA-lipid lamellar complex), the magnitude and anisotropy of the diffusion coefficient of the DNA fragments confined by the lipid bilayers are obtained for the first time. In the second, binary DNA-solvent system, the magnitude of the diffusion coefficient is found to decrease markedly as DNA concentration is increased from isotropic to cholesteric phase. In addition, the diffusion coefficient anisotropy measured within cholesteric domains in the phase coexistence region increases with concentration, and eventually reaches a high value in the cholesteric phase.

Effects of Natural Radiation, Photosynthetically Active Radiation and Artificial Ultraviolet Radiation-B on the Chloroplast Organization and Metabolism of Porphyra acanthophora var. brasiliensis (Rhodophyta, Bangiales)

We undertook a study of Porphyra acanthophora var. brasiliensis to determine its responses under ambient conditions, photosynthetically active radiation (PAR), and PAR+UVBR (ultraviolet radiation-B) treatment, focusing on changes in ultrastructure, and cytochemistry. Accordingly, control ambient samples were collected in the field, and two different treatments were performed in the laboratory. Plants were exposed to PAR at 60 mu mol photons m(-2) s(-1) and PAR+UVBR at 0.35 W m(-2) for 3 h per day during 21 days of in vitro cultivation. Confocal laser scanning microscopy analysis of the vegetative cells showed single stellate chloroplast in ambient and PAR samples, but in PAR+UVBR-exposed plants, the chloroplast showed alterations in the number and form of arms. Under PAR+UVBR treatment, the thylakoids of the chloroplasts were disrupted, and an increase in the number of plastoglobuli was observed, in addition to mitochondria, which appeared with irregular, disrupted morphology compared to ambient and PAR samples. After UVBR exposure, the formation of carpospores was also observed. Plants under ambient conditions, as well as those treated with PAR and PAR+UVBR, all showed different concentrations of enzymatic response, including glutathione peroxidase and reductase activity. In summary, the present study demonstrates that P. acanthophora var. brasiliensis shows the activation of distinct mechanisms against natural radiation, PAR and PAR+UVBR.

Schistosoma mansoni: In vitro schistosomicidal activity and tegumental alterations induced by piplartine on schistosomula

Schistosomiasis is one of the most important parasitic infections in humans that occur in many tropical and subtropical countries. Currently, the control of schistosomiasis rests with a single drug, praziquantel, which is effective against adult worms but not the larval stages. Recent studies have shown that piplartine, an amide isolated from plants of the genus Piper (Piperaceae), reveals interesting antischistosomal properties against Schistosoma mansoni adult worms. Here, we report the in vitro antischistosomal activity of piplartine on S. mansoni schistosomula of different ages (3 h old and 1, 3, 5, and 7 days old), and examine alterations on the tegumental surface of worms by means of confocal laser scanning microscopy. Piplartine at a concentration of 7.5 mu M caused the death of all schistosomula within 120 h. The lethal effect occurred in a dose-dependent manner and was also dependent on the age of the parasite. Microscopy observation revealed extensive tegumental destruction, including blebbing, granularity, and a shorter body length. This report provides the first evidence that piplartine is able to kill schistosomula of different ages and reinforce that piplartine is a promising compound that could be used for the development of new schistosomicidal agent. (c) 2012 Elsevier Inc. All rights reserved.

Mast cells act as phagocytes against the periodontopathogen Aggregatibacter Actinomycetemcomitans

Background: Evidence to date shows that mast cells play a critical role in immune defenses against infectious agents, but there have been no reports about involvement of these cells in eliminating periodontopathogens. In this study, the phagocytic ability of mast cells against Aggregatibacter actinomycetemcomitans compared with macrophages is evaluated. Methods: In vitro phagocytic assays were conducted using murine mast cells and macrophages, incubated with A. actinomycetemcomitans, either opsonized or not, with different bacterial load ratios. After 1 hour, cells were stained with acridine orange and assessed by confocal laser-scanning electronmicroscopy. Results: Phagocytic ability of murine mast cells against A. actinomycetemcomitans was confirmed. In addition, the percentage of mast cells with internalized bacteria was higher in the absence of opsonization than in the presence of opsonization. Both cell types showed significant phagocytic activity against A. actinomycetemcomitans. However, the percentage of mast cells with non-opsonized bacteria was higher than that of macrophages with opsonized bacteria in one of the ratios (1:10). Conclusions: This is the first report about the participation of murine mast cells as phagocytes against A. actinomycetemcomitans, mainly in the absence of opsonization with human serum. Our results may indicate that mast cells act as professional phagocytes in the pathogenesis of biofilmassociated periodontal disease

In vitro effects of Er,Cr:YSGG laser on dentine hypersensitivity. Dentine permeability and scanning electron microscopy analysis

The aim of the present study was to determine clinical parameters for the use of Er,Cr:YSGG laser in the treatment of dentine hypersensitivity. Two antagonist areas were determined as control and experimental areas for irradiation in 90 premolar roots. Each surface was conditioned with 24% EDTA (sub-group 1) and 35% phosphoric acid (sub-group 2) and irradiated with the following settings: 1) Er:YAG, 60 mJ, 2 Hz, defocused; groups 2 to 9: irradiation with Er,Cr:YSGG laser, 20 Hz, Z6 tip, 0% of air and water: 2) Er,Cr:YSGG 0.25 W; 3) 0.5 W; 4) 0.75 W; 5) 1.0 W; 6) 1.25 W, 7) 1.50 W, 8) 2 W; 9) 2 W. After irradiation, samples were immersed in methylene blue solution and included in epoxy resin to obtain longitudinal cuts. The images were digitalized and analyzed by computer software. Although the samples irradiated with Er:YAG laser showed less microleakage, sub-group 1 showed differences between the groups, differing statistically from groups 3, 6, and 9. The results of sub-group 2 showed that the mean values of Er:YAG samples showed a negative trend, however, no differences were detected between the groups. For scanning electron microscopy analysis, dentine squares were obtained and prepared to evaluate the superficial morphology. Partial closure of dentinal tubules was observed after irradiation with Er:YAG and Er,Cr:YSGG laser in the 0.25 and 0.50 W protocols. As the energy densities rose, open dentinal tubules, carbonization and cracks were observed. It can be concluded that none of the parameters were capable of eliminating microleakage, however, clinical studies with Er:YAG and Er,Cr:YSGG lasers should be conducted with the lowest protocols in order to determine the most satisfactory setting for dentine hypersensitivity.

Effect of pulse repetition rate and number of pulses in the analysis of polypropylene and high density polyethylene by nanosecond infrared laser induced breakdown spectroscopy

Pulse repetition rates and the number of laser pulses are among the most important parameters that do affect the analysis of solid materials by laser induced breakdown spectroscopy, and the knowledge of their effects is of fundamental importance for suggesting analytical strategies when dealing with laser ablation processes of polymers. In this contribution, the influence of these parameters in the ablated mass and in the features of craters was evaluated in polypropylene and high density polyethylene plates containing pigment-based PbCrO4. Surface characterization and craters profile were carried out by perfilometry and scanning electron microscopy. Area, volume and profile of craters were obtained using Taylor Map software. A laser induced breakdown spectroscopy system consisted of a Q-Switched Nd:YAG laser (1064 nm, 5 ns) and an Echelle spectrometer equipped with ICCD detector were used. The evaluated operating conditions consisted of 10, 25 and 50 laser pulses at 1, 5 and 10 Hz, 250 mJ/pulse (85 J cm(-2)), 2 mu s delay time and 6 mu s integration time gate. Differences in the topographical features among craters of both polymers were observed. The decrease in the repetition rate resulted in irregular craters and formation of edges, especially in polypropylene sample. The differences in the topographical features and ablated masses were attributed to the influence of the degree of crystallinity, crystalline melting temperature and glass transition temperature in the ablation process of the high density polyethylene and polypropylene. It was also observed that the intensities of chromium and lead emission signals obtained at 10 Hz were two times higher than at 5 Hz by keeping the number of laser pulses constant. (C) 2011 Elsevier B. V. All rights reserved.

Nd:YAG Laser Irradiation of Etched/Unetched Dentin Through an Uncured Two-step Etch-and-Rinse Adhesive and its Effect on Microtensile Bond Strength

Purpose: To evaluate whether Nd:YAG laser irradiation of etched and unetched dentin through an uncured adhesive affected the microtensile bond strength (pTBS). Materials and Methods: Flat dentin surfaces were created in 19 extracted human third molars. Adper Single Bond (SB) adhesive was applied over etched (groups 1 to 3) or unetched dentin (groups 4 to 6). The dentin was then irradiated with a Nd:YAG laser through the uncured adhesive, using 0.75 or 1 W power settings, except for the control groups (groups 1 and 4). The adhesive was light cured and composite crowns were built up. After 24 h, the teeth were sectioned into beams, with cross-sectional areas of 0.49 mm(2), and were stressed under tension. Data were statistically analyzed using two-way ANOVA and Tukey's test (alpha = 5%). Dentin surfaces of fractured specimens and the interfaces of untested beams were observed under scanning electron microscopy (SEM). Results: Acid etching, laser irradiation, and their interaction significantly affected bonding (p < 0.05). Laser irradiation did not improve bonding of etched dentin to resin (p > 0.05). However, higher pTBS means were found on unetched lased dentin (groups 5 and 6), but only in comparison to group 4, where neither lasing nor etching was performed. Groups 4 to 6 showed the lowest pTBS means among all groups tested (p < 0.05). Laser irradiation did not change the characteristics of the hybrid layers created, while solidification globules were observed on lased dentin surfaces under SEM. Conclusion: Laser irradiation of dentin through the uncured adhesive did not significantly improve the pTBS in comparison to the suggested manufacturer's technique.

Influence of prior 810-nm-diode intracanal laser irradiation on hydrophilic resin-based sealer obturation

Dentin wall structural changes caused by 810-nm-diode laser irradiation can influence the sealing ability of endodontic sealers. The objective of this study was to evaluate the apical leakage of AH Plus and RealSeal resin-based sealers with and without prior diode laser irradiation. Fifty-two single-rooted mandibular premolars were prepared and divided into 4 groups, according to the endodontic sealer used and the use or non-use of laser irradiation. The protocol for laser irradiation was 2.5W, continuous wave in scanning mode, with 4 exposures per tooth. After sample preparation, apical leakage of 50% ammoniacal silver nitrate impregnation was analyzed. When the teeth were not exposed to irradiation, the Real Seal sealer achieved the highest scores, showing the least leakage, with significant differences at the 5% level (Kruskal-Wallis test, p = 0.0004), compared with AH Plus. When the teeth were exposed to the 810-nm-diode laser irradiation, the sealing ability of AH Plus sealer was improved (p = 0282). In the Real Seal groups, the intracanal laser irradiation did not interfere with the leakage index, showing similar results in the GRS and GRSd groups (p = 0.1009).

Thermal alteration and morphological changes of sound and demineralized primary dentin after Er:YAG laser ablation

The purpose of this study was to assess the influence of Er:YAG laser pulse repetition rate on the thermal alterations occurring during laser ablation of sound and demineralized primary dentin. The morphological changes at the lased areas were examined by scanning electronic microscopy (SEM). To this end, 60 fragments of 30 sound primary molars were selected and randomly assigned to two groups (n = 30); namely A sound dentin (control) and B demineralized dentin. Each group was divided into three subgroups (n = 10) according to the employed laser frequencies: I4 Hz; II6 Hz, and III10 Hz. Specimens in group B were submitted to a pH-cycling regimen for 21 consecutive days. The irradiation was performed with a 250 mJ pulse energy in the noncontact and focused mode, in the presence of a fine water mist at 1.5 mL/min, for 15 s. The measured temperature was recorded by type K thermocouples adapted to the dentin wall relative to the pulp chamber. Three samples of each group were analyzed by SEM. The data were submitted to the nonparametric Kruskal-Wallis test and to qualitative SEM analysis. The results revealed that the temperature increase did not promote any damage to the dental structure. Data analysis demonstrated that in group A, there was a statistically significant difference among all the subgroups and the temperature rise was directly proportional to the increase in frequency. In group B, there was no difference between subgroup I and II in terms of temperature. The superficial dentin observed by SEM displayed irregularities that augmented with rising frequency, both in sound and demineralized tissues. In conclusion, temperature rise and morphological alterations are directly related to frequency increment in both demineralized and sound dentin. Microsc. Res. Tech., 2011. (c) 2011 Wiley Periodicals, Inc.

Screening of CO2 Laser (10.6 mu m) Parameters for Prevention of Enamel Erosion

Objective: The aim of this study was to screen CO2 laser (10.6 mu m) parameters to increase enamel resistance to a continuous-flow erosive challenge. Background data: A new clinical CO2 laser providing pulses of hundreds of microseconds, a range known to increase tooth acid-resistance, has been introduced in the market. Methods: Different laser parameters were tested in 12 groups (n = 20) with varying fluences from 0.1 to 0.9 J/cm(2), pulse durations from 80 to 400 mu s and repetition rates from 180 to 700 Hz. Non-lased samples (n = 30) served as controls. All samples were eroded by exposure to hydrochloric acid (pH 2.6) under continuous acid flow (60 mu L/min). Calcium and phosphate release into acid was monitored colorimetrically at 30 sec intervals up to 5 min and at 1 min intervals up to a total erosion time of 15 min. Scanning electron microscopic (SEM) analysis was performed in lased samples (n = 3). Data were statistically analysed by one-way ANOVA (p < 0.05) and Dunnett's post-hoc tests. Results: Calcium and phosphate release were significantly reduced by a maximum of 20% over time in samples irradiated with 0.4 J/cm(2) (200 mu s) at 450 Hz. Short-time reduction of calcium loss (<= 1.5 min) could be also achieved by irradiation with 0.7 J/cm(2) (300 mu s) at 200 and 300 Hz. Both parameters revealed surface modification. Conclusions: A set of CO2 laser parameters was found that could significantly reduce enamel mineral loss (20%) under in vitro erosive conditions. However, as all parameters also caused surface cracking, they are not recommended for clinical use.

Thermal effects and morphological aspects of human dentin surface irradiated with different frequencies of Er:YAG laser

This study reports the effects on micromorphology and temperature rise in human dentin using different frequencies of Er:YAG laser. Sixty human dentin fragments were randomly assigned into two groups (n = 30): carious or sound dentin. Both groups were divided into three subgroups (n = 10), according to the Er:YAG laser frequency used: 4, 6, or 10 Hz (energy: 200 mJ; irradiation distance: 12 mm; and irradiation time: 20 s). A thermocouple adapted to the tooth fragment recorded the initial temperature value (degrees C); then, the temperature was measured after the end of the irradiation (20 s). Morphological analysis was performed using images obtained with scanning electron microscope. There was no difference between the temperatures obtained with 4 and 6 Hz; the highest temperatures were achieved with 10 Hz. No difference was observed between carious and sound dentin. Morphological analyses revealed that all frequencies promoted irregular surface in sound dentin, being observed more selectively ablation especially in intertubular dentin with tubule protrusion. The caries dentin presented flat surface for all frequencies used. Both substrates revealed absence of any signs of thermal damage. It may be concluded that the parameters used in this study are capable to remove caries lesion, having acceptable limits of temperature rise and no significant morphological alterations on dentin surface. Microsc. Res. Tech. 2012. (c) 2012 Wiley Periodicals, Inc.