Composite filling removal with erbium:yttrium-aluminum-garnet laser: morphological analyses

Considering the increase in esthetic restorative materials and need for improvement in unsatisfactory restoration substitution with minimal inadvertent removal of healthy tissues, this study assessed the efficacy of erbium:yttrium-aluminum-garnet (Er:YAG) laser for composite resin removal and the influence of pulse repetition rate on the morphological analyses of the cavity by scanning electron microscope. Composite resin fillings were placed in cavities (1.0 mm deep) prepared in bovine teeth, and the 75 specimens were randomly assigned to five groups according to the technique used for composite filling removal (high-speed diamond bur, group I, as a control, and Er:YAG laser, 250 mJ output energy and 80 J/cm(2) energy density, using different pulse repetition rates: group II, 2 Hz; group III, 4 Hz; group IV, 6 Hz; group V, 10 Hz). After the removal, the specimens were split in the middle, and we analyzed the surrounding and deep walls to check for the presence of restorative material. The estimation was qualitative. The surfaces were examined with a scanning electron microscope. The results revealed that the experimental groups presented bigger amounts of remaining restorative material. The scanning electron microscopy (SEM) analyses showed irregularities of the resultant cavities of the experimental groups that increased proportionally with increase in repetition rate.

Shear Bond Strength of A Sealant to Contaminated-Enamel Surface: Influence of Erbium : Yttrium-Aluminum-Garnet Laser Pretreatment

Salivary contamination is one of the factors that can disturb the sealing process and interfere in the longevity of pit and fissure sealants. Erbium : yttrium-aluminum-garnet (Er : YAG) laser could influence the bond strength of enamel and increase the acid resistance. To evaluate the influence of Er : YAG laser on the shear bond strength of a sealant to a salivary contaminated enamel surface. Twenty-four third molars had the roots sectioned 2 mm coronal to the cementoenamel junction. The crowns were mesiodistally sectioned providing 48 halves that were embedded in polyester resin. Enamel was flattened and a 2-mm diameter bonding area was demarcated. Specimens were randomly assigned to two groups according to the superficial pretreatment-37% phosphoric acid (A) and Er : YAG laser (80 mJ/2 Hz) + phosphoric acid (L), which were subdivided into two groups (N = 12), without salivary contamination (C) and with salivary contamination (SC). To contaminate the specimens, 0.25 mL of human fresh saliva was applied for 20 seconds and then dried. Fluroshield sealant was applied in all specimens. After storage, shear bond strength of samples were tested in a universal testing machine. Means in MPa were: AC-14.61 (+/- 2.52); ASC-6.66 (+/- 2.34); LC-11.91 (+/- 1.34); and LSC-2.22 (+/- 0.66). Statistical analysis revealed that surfaces without salivary contamination and with acid treatment had the highest mean (p < 0.05). The group with salivary contamination treated by Er : YAG laser followed by phosphoric acid application presented the lowest bond values (p < 0.05). The phosphoric acid etching under dry condition yielded better bonding performance. Er : YAG laser was not able to increase the effectiveness of conventional acid etching of enamel in the bond of sealants in both dry and wet conditions. Under the conditions of this study, the conventional etching protocol (phosphoric acid without salivary contamination) is still preferable to laser-conditioning enamel surface prior to sealant application.

Radiopacity Evaluation of Calcium Aluminate Cement Containing Different Radiopacifying Agents

Introduction: The aim of this study was to evaluate the radiopacity of calcium aluminate cement (Endo Binder) with 3 different radiopacifiers (bismuth oxide, zinc oxide, or zirconium oxide) in comparison with gray mineral trioxide aggregate (GMTA), white MTA, and dental structures (enamel and dentin). Methods: Eighteen test specimens of each cement with thicknesses of 0.5, 1.0, 1.5, 2.0, 2.5, and 3.0 mm (n = 3) were made by using a stainless steel matrix and were adapted to a standardizing device (8 x 7 cm) with a graduated aluminum stepwedge varying from 2.0-16.0 mm in thickness. To compare the radiopacity of the cements with that of dental structures, slices of first molars with a thickness increasing from 0.5-3.0 mm were obtained and placed on the standardizing device. One occlusal radiograph for each tested cement was taken, with exposure time of 0.1 seconds and focus-film distance of 20 cm. Films were processed in an automatic device, and the mean radiopacity values were obtained by using a photodensitometer. Results: Mean values showed that the thicker the specimen was, the greater was its radiopacity. Only EndoBinder + bismuth oxide (EBBO) and GMTA demonstrated radiopacity values greater than 3.0 mm of the aluminum scale for all thicknesses. When zinc oxide was used as radiopacifier agent, EndoBinder only reached the desired radiopacity with a thickness of 2.0 mm, and with zirconium oxide it was 2.5 mm. Conclusions: Bismuth oxide was the most efficient radiopacifier for EndoBinder, providing adequate radiopacity in all studied thicknesses, as recommended by ISO 6876, being similar to GMTA. (J Endod 2011;37: 67-71)

Nitric oxide detection in cell culture exposed to LPS after Er:YAG laser irradiation

P>Aim To evaluate in vitro the effect of calcium hydroxide [Ca(OH)(2)] and Er:YAG laser on bacterial endotoxin [also known as lipopolysaccharide (LPS)] as determined by nitric oxide (NO) detection in J774 murine macrophage cell line culture. Methodology Samples of LPS solution (50 mu gmL-1), Ca(OH)(2) suspension (25 mg mL-1) and LPS suspension with Ca(OH)(2) were prepared. The studied groups were: I - LPS (control); II - LPS + Ca(OH)(2); III - LPS + Er:YAG laser (15 Hz 140 mJ); IV - LPS + Er:YAG laser (15 Hz 200 mJ); V - LPS + Er:YAG laser (15 Hz 250 mJ), VI - Pyrogen-free water; VII - Ca(OH)(2). Murine macrophage J774 cells were plated and 10 mu L of the samples were added to each well. The supernatants were collected for NO detection by the Griess reaction. Data were analysed statistically by one-way anova and Tukey`s test at 5% significance level. Results The mean and SE (in mu mol L-1) values of NO release were: I - 10.48 +/- 0.58, II - 6.41 +/- 0.90, III - 10.2 +/- 0.60, IV - 8.35 +/- 0.40, V - 10.40 +/- 0.53, VI - 3.75 +/- 0.70, VII - 6.44 +/- 0.60; and the values for the same experiment repeated after 1 week were: I - 21.20 +/- 1.50, II - 9.10 +/- 0.60, III - 19.50 +/- 1.00, IV - 18.50 +/- 0.60, V - 21.30 +/- 0.90, VI - 2.00 +/- 0.20, VII - 6.80 +/- 1.70. There was no significant difference (P > 0.05) between the control and the laser-treated groups (III, IV and V), or comparing groups II, VI and VII to each other (P > 0.05). Group I had significantly higher NO release than group II (P < 0.05). Groups II and VI had similar NO release (P > 0.05). Conclusions Calcium hydroxide inactivated the bacterial endotoxin (LPS) whereas none of the Er:YAG laser parameter settings had the same effectiveness.

Bond durability in erbium:yttrium-aluminum-garnet laser-irradiated enamel

This study sought to evaluate the influence of thermocycling and water storage on the microtensile bond strength of composite resin bonded to erbium:yttrium-aluminum-garnet (Er:YAG)-irradiated and bur-prepared enamel. Eighty bovine incisors were selected and sectioned. Specimens were ground to produce a flat enamel surface. Samples were randomly assigned according to cavity preparation device: (I) Er:YAG laser and (II) high-speed turbine, and were subsequently restored with composite resin. They were subdivided according to the duration of water storage (WS)/number of thermocycles (TCs): 24 h WS/no TCs; 7 days WS/500 TCs; 1 month WS/2,000 TCs; 6 months WS/12,000 TCs. The teeth were sectioned into 1.0 mm(2)-thick slabs and subjected to tensile stress in a universal testing machine. Data were submitted to two-way analysis of variance (ANOVA) and Tukey`s test at a 0.05 significance level. The different periods of water storage and thermocycling did not influence the microtensile bond strength (A mu TBS) values in the Er:YAG laser-prepared groups. In bur-prepared enamel, the group submitted to 12,000 TCs/6 months` WS (IID) showed a significant decrease in bond strength values when compared to the group stored in water for 24 h and not submitted to thermocycling (IIA), but values were statistically similar to those obtained in all Er:YAG laser groups and in the bur- prepared groups degraded with 500 TCs/1 week WS (IIB) or 2,000 TCs/1 month WS (IIC). It may be concluded that adhesion of an etch-and-rinse adhesive to Er:YAG laser-irradiated enamel was not affected by the methods used to simulate degradation of the adhesive interface and was similar to adhesion in the bur-prepared groups in all periods of water storage and thermocycling.

Endodontic Chelators Induce Nitric Oxide Expression by Murine-cultured Macrophages

Introduction: Endodontic chelators may extrude to apical tissues during instrumentation activating cellular events on periapical tissues. This study assessed in vitro the expression of nitric oxide (NO) concentrations by murine peritoneal macrophages after contact with MTAD (Dentsply/Tulsa, Tulsa, OK), Tetraclean (Ogna Laboratori Farmaceutici, Muggio, Italy), Smear Clear (Sybron Endo, Orange, CA), and EDTA (Biodinamica, Ibipora, PR, Brazil). Methods: Macrophage cells were obtained from Swiss mice after peritoneal lavage. Chelators were diluted in distilled water obtaining 12 concentrations, and MTT assay identified the concentrations, per group, displaying the highest cell viability (analysis of variance, p < 0.01). Selected concentrations were tested for NO expression using Griess reaction. Culture medium and lipopolysaccharide (LPS) were used as controls. Results: Analysis of variance and Tukey tests showed that all chelators displayed elevated NO concentrations compared with the negative control (p < 0.01). MTAD induced the lowest NO expression, followed by Tetraclean, EDTA, and Smear Clear. No difference was observed between MTAD and Tetraclean (p > 0.01), Tetraclean and EDTA (p > 0.01), and EDTA and Smear Clear (p > 0.01). LPS ranked similar to both EDTA and Smear Clear (p > 0.01). Conclusion: The tested endodontic chelators displayed severe proinflammatory effects on murine-cultured macrophages. Citric acid-based solutions induce lower No release than EDTA-based irrigants. (J Endod 2009;35:824-828)

Trigeminal nitric oxide synthase expression correlates with new bone formation during distraction osteogenesis

Nitric oxide synthase (NOS) has been reported to be involved with both bone healing and bone metabolism. The aim of this study was to test the null hypothesis that there is no correlation between new bone formation during mandibular distraction osteogenesis and NOS expression in the trigeminal ganglion of rats. Newly formed tissue during distraction osteogenesis and trigeminal NOS expression measured by the NADPH-diaphorase (NADPH-d) reaction were evaluated in 72 male Wistar rats by histomorphometric and histochemical methods. In animals submitted to 0.5 mm/day distraction osteogenesis, the percentage of bone tissue was higher in the basal area of the mandibles compared with the center and significantly increased through the experimental periods (P < 0.05). At the sixth postoperative week, the difference in bone formation between the continuous and acute distraction osteogenesis groups was the highest. Significant correlation between new bone formation by distraction osteogenesis and NADPH-d-reactive neurons was found, varying according to neuronal cell size (r = -0.6, P = 0.005, small cells strongly stained; r = 0.5, P = 0.018, large cells moderately stained). The results suggest that NOS may play a role in the bone healing process via neurogenic pathways, and the phenomenon seems to be neuronal cell morphotype-dependent. Further studies are now warranted to investigate the mechanistic link between the expression of trigeminal NOS and mandibular new bone formation by distraction osteogenesis.

Endothelial and inducible nitric oxide synthases in oocytes of cattle

Nitric oxide (NO) is a chemical messenger generated by the activity of the nitric oxide synthases (NOS). The NOS/NO system appears to be involved in oocyte maturation, but there are few studies on gene expression and protein activity in oocytes of cattle. The present study aimed to investigate gene expression and protein activity of NOS in immature and in vitro matured oocytes of cattle. The influence of pre-maturation culture with butyrolactone I in NOS gene expression was also assessed. The following experiments were performed: (1) detection of the endothelial (eNOS) and inducible (iNOS) isoforms in the ovary by immunohistochemistry; (2) detection of eNOS and iNOS in the oocytes before and after in vitro maturation (W) by immunofluorescence; (3) eNOS and iNOS mRNA and protein in immature and in vitro matured oocytes, with or without pre-maturation, by real time PCR and Western blotting, respectively; and (4) NOS activity in immature and in vitro matured oocytes by NADPH-diaphorase. eNOS and iNOS were detected in oocytes within all follicle categories (primary, secondary and tertiary), and other compartments of the ovary and in the cytoplasm of immature and in vitro matured oocytes. Amount of mRNA for both isoforms decreased after IVM but was maintained after pre-maturation culture. The NOS protein was detected in immature (pre-mature or not) and was still detected in similar amount after pre-maturation and maturation for both isoforms. NOS activity was detected only in part of the immature oocytes. In conclusion, isoforms of NOS (eNOS and iNOS) are present in oocytes of cattle from early folliculogenesis up to maturation; in vitro maturation influences amount of mRNA and NOS activity. (C) 2009 Elsevier B.V. All rights reserved.

Influence of nitric oxide during maturation on bovine oocyte meiosis and embryo development in vitro

The effect of s-nitroso-N-acetyl-1,1-penicillamine (SNAP, a nitric oxide donor) during in vitro maturation (IVM) on nuclear maturation and embryo development was investigated. The effect of increasing nitric oxide (NO) during prematuration or maturation, or both, on embryo development was also assessed. 10(-3) M SNAP nearly blocked oocytes reaching metaphase II (MII) (7%, P < 0.05) while 10(-5) M SNAP showed intermediate proportions (55%). For 10(-7) M SNAP and controls (without SNAP), MII percentages were similar (72% for both, P > 0.05), but superior to the other treatment groups (P < 0.05). Blastocyst development, however, was not affected (38% for all treatments, P < 0.05). TUNEL-positive cells in hatched blastocysts (Day 9) increased when IVM included 10(-5) M SNAP (8 v. 3 to 4 cells in the other treatments, P > 0.05), without affecting total cell numbers (240 to 291 cells, P > 0.05). When oocytes were prematured followed by IVM with or without 10(-7) M SNAP, during either culture period or both, blastocyst development was similar (26 to 40%, P > 0.05). When SNAP was included during both prematuration and IVM, the proportion of Day 9 hatched embryos increased (28% v. 14 to 19% in the other treatments, P < 0.05). Apoptotic cells, however, increased when SNAP was included (6 to 10 cells) in comparison to prematuration and maturation without SNAP (3 cells, P < 0.05). NO may be involved in meiotic progression and apoptosis during embryo development.

Dielectric properties and physical features of phosphate glasses containing iron oxide

Microwave techniques were applied to the study of dielectric properties of phosphate glasses on the basis of contributions from permanent and induced dipolar polarization of local structural units interacting with the electrical component of the electromagnetic radiation. The dielectric constant of the selected glass system (100-x)(50P(2)O(5)center dot 25Li(2)O center dot 25Na(2)O)center dot xFe(2)O(3), where 0 <= x <= 21 is in mol%, was measured using a microwave setup assembled to measure the phase shift of the standing wave pattern produced by the insertion of the sample. It is shown that the Fe2+ ions contribute effectively to the dielectric constant, as expected from the interactions of the dipoles of the local charge compensation pairs with the microwave radiation. However, there is the possibility of occurrence of some ions Fe3+, in general, at low iron content, which reinforces the glass structure and, therefore, decreases the dielectric constant. There is a gradual conversion from Fe3+ to Fe2+ as the iron ions increases. This is possibly the reason of the anomaly in the dielectric constant values observed in the results. These assumptions can be checked by results of electronic paramagnetic resonance (EPR) and optical absorption (OA). The dielectric constant of the glasses studied in this work was found to increase with the temperature in the range of 25-330 degrees C. (C) 2007 Elsevier B.V. All rights reserved.

Aluminum as an endocrine disruptor in female Nile tilapia (Oreochromis niloticus)

The effects of aluminum on plasma ion, lipid, protein and steroid hormone concentration were evaluated in Oreochromis niloticus broodstock females. Lipid and protein concentrations from the gonads and liver were also measured Experiments were performed at neutral and acidic water pH Four groups of fish were tested for 96 h. 1) control conditions at neutral water pH, 2) control conditions at acidic water pH (CTR-Ac). 3) aluminum at neutral water pH (Al-N), and 4) aluminum at acidic water pH (Al-Ac) Aluminum and acidic water pH exposure caused no ionoregulatory disturbances Total lipid concentration increased in the mature gonads and decreased in the liver, suggesting an acceleration of lipid mobilization to the ovaries in animals exposed to aluminum However, a decreased protein concentration in ovaries was also observed Exposure of control fish to acidic water pH caused an increased concentration of plasma 17 alpha-hydroxyprogesterone However, females exposed to aluminum at acidic water pH showed a decreased of plasma 17 alpha-hydroxyprogesterone and cortisol. No differences in plasma 17 beta-estradiol were observed The physiological mechanisms underlying the disturbances observed are discussed focusing on reproduction We suggest that aluminum can be considered an endocrine disrupting compound in mature O. mloticus females (C) 2010 Elsevier Inc. All rights reserved

Nitrosyl ethylenediaminetetraacetate ruthenium(II) complex promotes cellular growth and could be used as nitric oxide donor in plants

Over recent years nitric oxide (NO) not only has appeared as an important endogenous signaling molecule in plants and as a mediator in many developmental and physiological processes, but has also received recognition as a plant hormone. The impressive recent achievements in elucidating the role of NO in plants have come about by the application of NO donors. The aim herein was to study the effects of the different NO donors, sodium nitroprusside (SNP) and the nitrosyl ethylenediaminetetraacetate ruthenium(II) ([Ru(NO)(Hedta)]) complex on cellular growth in embryogenic suspension cultures of Araucaria angustifolia. Appraisal of our data revealed that [Ru(NO)(Hedta)] stimulated about 60% of cellular growth in embryogenic suspension cultures of A. angustifolia, with results similar to those observed with the SNP donor. Nevertheless, application of the NO scavenger PTIO (2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide) inhibited this cellular growth in both. Cellular growth was correlated with an increase in endogenous NO levels after 21 days of culture, especially in treatments with NO donors. Our results demonstrated that the [Ru(NO)Hedta] complex could possibly be used as a NO donor in plants. (C) 2010 Elsevier Ireland Ltd. All rights reserved.

In vivo blockade of Ca(+2)-dependent nitric oxide synthases impairs expressions of L-selectin and PECAM-1

Interactions of leukocytes with endothelium play a role for the immune system modulated by endogenous agents, such as glucocorticoids and nitric oxide (NO). Glucocorticoids inhibit leukocyte-endothelial interactions whereas the role of NO is still controversial. In this study, the activity of Ca(+2)-dependent nitric oxide synthases was in vivo blocked in male Wistar rats by given L-NAME, 20 mg kg(-1) for 14 days dissolved in drinking water and expression of adhesion molecules involved in leukocyte-endothelial interactions was investigated. Expressions of L-selectin and PECAM-I in peripheral leukocytes and PECAM-1 in endothelial cells were reduced by L-NAME treatment. Only L-selectin expression was controlled at transcriptional levels. These effects were not dependent on endogenous glucocorticoids, as corticosterone levels were not altered in NAME-treated rats. Our results show that NO, produced at physiological levels, controls expression of constitutive adhesion molecules expressions in cell membranes by different mechanisms of action. Published by Elsevier Inc.

Apolipoprotein E genotype is related to nitric oxide production in platelets

The presence of the, 4 allele of apolipoprotein E (APOE) is considered a risk factor for sporadic Alzheimer`s disease (AD). Our recent data demonstrated that the systemic modulation of oxidative stress in platelets and erythrocytes is disrupted in aging and AD. In this study, the relationship between APOE genotype and oxidative stress markers, both in AD patients and controls, was evaluated. The AD group showed an increase in the content of thiobarbituric acid-reactive substances (TBARS) and in the activities of nitric oxide synthase (NOS) and Na, K-ATPase, when compared to controls. Both groups had a similar cGMP content and superoxide dismutase activity. APOE epsilon 4 allele carriers showed higher NOS activity than non-carriers. These results suggest a possible influence of APOE genotype on nitric oxide (NO) production that might enhance the effects of age-related specific factor(s) associated with neurodegenerative disorders. Copyright (C) 2008 John Wiley & Sons, Ltd.

Chronic ouabain treatment increases the contribution of nitric oxide to endothelium-dependent relaxation

The aim of this study was to analyze the contribution of nitric oxide, prostacyclin and endothelium-dependent hyperpolarizing factor to endothelium-dependent vasodilation induced by acetylcholine in rat aorta from control and ouabain-induced hypertensive rats. Preincubation with the nitric oxide synthase inhibitor N-omega-nitro-L-arginine methyl esther (L-NAME) inhibited the vasodilator response to acetylcholine in segments from both groups but to a greater extent in segments from ouabain-treated rats. Basal and acetylcholine-induced nitric oxide release were higher in segments from ouabain-treated rats. Preincubation with the prostacyclin synthesis Inhibitor tranylcypromine or with the cyclooxygenase inhibitor indomethacin inhibited the vasodilator response to acetylcholine in aortic segments front both groups. The Ca(2+)-dependent potassium channel blocker charybdotoxin inhibited the vasodilator response to acetylcholine only In segments from control rats. These results indicate that hypertension induced by chronic ouabain treatment is accompanied by increased endothelial nitric oxide participation and impaired endothelium-dependent hyperpolarizing factor contribution In acetylcholine-induced relaxation. These effects might explain the lack of effect of ouabain treatment oil acetylcholine responses in rat aorta.