In vitro evaluation of apical microleakage using different root-end filling materials

The objective of this study was to evaluate the apical leakage of retrograde cavities filled with Portland Cement (Concrebrás S/A-MG-Brazil), ProRoot MTA™ (Dentsply International, Johnson City, TN, USA) and Sealapex (Kerr Corporation, Orange, California, USA) with addition of zinc oxide (Odahcam Herpo Produtos Dentários Ltda, Rio de Janeiro, RJ, Brazil). Forty-two extracted single-rooted human teeth were decoronated and used for this study. The root canals were instrumented at 1.0mm short of the apical foramen using the step-back technique to an apical ISO size 60. The roots were obturated with gutta-percha points and sealer Sealapex (Kerr Corporation-USA) and then 3mm of each root apex was sectioned at a 90° angle. Ultrasonic retrograde preparation was performed with a diamond tip to 3mm depth and the roots were randomly divided into 3 groups according to the filling material: G1-Portland, G2-ProRoot MTA, G3- Sealapex zinc oxide-added cement. The root surfaces were covered with nail varnish up to 2mm from the apical foramen, immersed in simulated tissue fluid for 30 days, and then immersed in 0.2% Rhodamine B solution for 24 hours for evaluation of marginal leakage. The results showed mean leakage of 0.75, 0.35 and 0.35 for groups 1, 2 and 3, respectively; however, Kruskal-Wallis test revealed that there was no statistically significant difference among the results (p>0.05).

In vitro evaluation of the precision of working casts for implant-supported restoration with multiple abutments

Objective: The purpose of this study was to compare the accuracy of two working cast fabrication techniques using strain-gauge analysis. Methods: Two working cast fabrication methods were evaluated. Based on a master model, 20 working casts were fabricated by means of an indirect impression technique using polyether after splinting the square transfer copings with acrylic resin. Specimens were assigned to 2 groups (n=10): Group A (GA): type IV dental stone was poured around the abutment analogs in the conventional way; Group B (GB), the dental stone was poured in two stages. Spacers were used over the abutment analogs (rubber tubes) and type IV dental stone was poured around the abutment analogs in the conventional way. After the stone had hardened completely, the spacers were removed and more stone was inserted in the spaces created. Six strain-gauges (Excel Ltd.), positioned in a cast bar, which was dimensionally accurate (perfect fit) to the master model, recorded the microstrains generated by each specimen. Data were analyzed statistically by the variance analysis (ANOVA) and Tukey's test (α= 5%). Results: The microstrain values (με) were (mean±SD): GA: 263.7±109.07με, and GB: 193.73±78.83με. Conclusion: There was no statistical difference between the two methods studied.

In vitro evaluation of surface roughness of 4 resin composites after the toothbrushing process and methods to recover superficial smoothness.

OBJECTIVE: This study evaluated the efficiency of repolishing, sealing with surface sealant, and the joining of both in decreasing the surface roughness of resin-based composites after a toothbrushing process. METHOD AND MATERIALS: Ten specimens of each composite (Alert, Z100, Definite, and Prodigy Condensable), measuring 2 mm in thickness and 4 mm in diameter, were made and submitted to finishing and polishing processes on both sides of the specimens using the Sof-Lex system. The specimens were then subjected to toothbrushing (30,000 cycles), and surface roughness (Ra) was analyzed with a Surfcorder SE 1700 profilometer. The upper surface of each composite was etched with 37% phosphoric acid, and the surface-penetrating sealant Protect-it was applied on 1 surface. The roughness of these surfaces was again measured. On the other side, the surface of the specimen was repolished, and the efficiency of this procedure was measured using the profilometer. The surface roughness resulting from the joining of the 2 methods was verified by applying, in the final stage, the surface-penetrating sealant on the repolished surface. Data were analyzed with analysis of variance and Tukey test (P <.05). RESULTS: Results showed that the lowest surface roughness values were obtained for Definite, Z100, and Prodigy Condensable after the repolishing process and after the repolishing plus sealing. For Alert, the joining of repolishing plus sealing promoted the lowest values of surface roughness. CONCLUSION: Of the resin-based composites, Alert demonstrated the highest values of surface roughness in all the techniques tested.

In vitro evaluation of the abrasiveness of a commercial low-abrasive dentifrice and an experimental dentifrice containing vegetable oil

Toothpastes usually contain detergents, humectants, water colorant, fluoride and thickeners (e.g. silica). Tooth wear has a multi-factorial etilology and the use of abrasive dentifrices is related to abrasion of dental tissues during toothbrushing. This study evaluated in vitro the abrasiveness of a commercial silica gel low-abrasive dentrifice compared to an experimental dentifrice containing vegetable (almond) oil. Distilled water served as a control group. Acrylic specimens (8 per group) were submitted to simulated toothbrushing with slurries of the commercial dentifrice experimental dentifrice, almond oil and water in an automatic brushing machine programmed to 30,000 brush strokes for each specimen which is equivalent to 2 years of manual toothbrushing. Thereafter, surface roughness (Ra) of the specimens was analyzed with a Surfcorder SE 1700 profilometer. Data were analyzed statistically by ANOVA and Tukey's test at 5% significance level. There was no statistically significant differences (p>0.05) in the surface roughness after brushing with water almond oil experimental dentifrice. The commercial dentifrice produced rougher surfaces compared to the control and abrasive free products (p<0.05). Further studies are necessary in confirm the potential benefits of using vegetable oil in toothpaste as an alternative in abrasives in an attempt to minimize the tooth wear caused by toothbrushing.

In vitro assessment of pulp chamber temperature of different teeth submitted to dental bleaching associated with LED/laser and halogen lamp appliances

This study sought to assess the pulp chamber temperature in different groups of human teeth that had been bleached using hydrogen peroxide gel activated with halogen lamps or hybrid LED/laser appliances. Four groups of ten teeth (maxillary central incisors, mandibular incisors, mandibular canines, and maxillary canines) were used. A digital thermometer with a K-type thermocouple was placed inside pulp chambers that had been filled with thermal paste. A 35% hydrogen peroxide-based red bleaching gel was applied to all teeth and photocured for a total of three minutes and 20 seconds (five activations of 40 seconds each), using light from an LED/laser device and a halogen lamp. The temperatures were gauged every 40 seconds and the data were analyzed by three-way ANOVA, followed by Tukey's test. Regardless of the light source, statistically significant differences were observed between the groups of teeth. The mean temperature values (±SD) were highest for maxillary central incisors and lowest for mandibular canines. The halogen lamp appliance produced more pulp chamber heating than the LED/laser appliance. The increase in irradiation time led to a significant increase in temperature.

In vitro-evaluation of secondary caries formation around restoration.

The objective of this in vitro study was to evaluate demineralization around restorations. Class V preparations were made on the buccal and lingual surfaces of each tooth. TPH (Group 1), Fuji II LC (Group 2), Tetric (Group 3), Dyract (Group 4), GS 80 (Group 5) and Chelon Fil (Group 6) were randomly placed in equal numbers of teeth. The teeth were submitted to a pH-cycling model associated with a thermocycling model. Sections were made and the specimens were examined for the presence of demineralization under polarized light microscopy. Demineralization was significantly reduced with Chelon Fil (Group 6). Furthermore, a similar inhibitory effect on the development of demineralization was observed in Groups 2, 4 and 5.

In vitro evaluation of the effect of natural orange juices on dentin morphology

The patient's diet has been considered an important etiological factor of dentin hypersensitivity. The frequent ingestion of acidic substances can promote the loss of dental structure or remove the smear layer. The purpose of this study was to evaluate the degree of smear layer removal and dentinal tubules exposure by different natural orange juices. Extracted human teeth were submitted to manual scaling in order to develop the smear layer. Seventy dentin samples were obtained and distributed into the following groups: Control, lime orange, lime, valência orange, navel orange, mandarin, and tangerine. Each group included 2 methods of application: Topical and topical + friction. After preparation for SEM analysis, photomicrographs were assessed by a blind calibrated examiner using an index system. The Kruskal-Wallis test indicated a significant influence of the orange juices on smear layer removal. Significant difference was observed between navel orange, valência orange, mandarin and the control group (p < 0.05). These orange juices resulted in greater removal of the smear layer and greater opening of dentinal tubules. The comparison between the application methods for each group using the Mann-Whitney test showed that friction increased smear layer removal significantly only for lime orange and lime. The data suggest that certain natural orange juices are more effective in terms of smear layer removal and dentinal tubules exposure than others.

Human eosinophil adhesion and degranulation stimulated with eotaxin and RANTES in vitro: Lack of interaction with nitric oxide

Background: Airway eosinophilia is considered a central event in the pathogenesis of asthma. The toxic components of eosinophils are thought to be important in inducing bronchial mucosal injury and dysfunction. Previous studies have suggested an interaction between nitric oxide (NO) and chemokines in modulating eosinophil functions, but this is still conflicting. In the present study, we have carried out functional assays (adhesion and degranulation) and flow cytometry analysis of adhesion molecules (VLA-4 and Mac-1 expression) to evaluate the interactions between NO and CC-chemokines (eotaxin and RANTES) in human eosinophils. Methods: Eosinophils were purified using a percoll gradient followed byimmunomagnetic cell separator. Cell adhesion and degranulation were evaluated by measuring eosinophil peroxidase (EPO) activity, whereas expression of Mac-1 and VLA-4 was detected using flow cytometry. Results: At 4 h incubation, both eotaxin (100 ng/ml) and RANTES (1000 ng/ml) increased by 133% and 131% eosinophil adhesion, respectively. L-NAME alone (but not D-NAME) also increased the eosinophil adhesion, but the co-incubation of L-NAME with eotaxin or RANTES did not further affect the increased adhesion seen with chemokines alone. In addition, L-NAME alone (but not D-NAME) caused a significant cell degranulation, but it did not affect the CC-chemokine-induced cell degranulation. Incubation of eosinophils with eotaxin or RANTES, in absence or presence of L-NAME, did not affect the expression of VLA-4 and Mac-1 on eosinophil surface. Eotaxin and RANTES (100 ng/ml each) also failed to elevate the cyclic GMP levels above baseline in human eosinophils. Conclusion: Eotaxin and RANTES increase the eosinophil adhesion to fibronectin-coated plates and promote cell degranulation by NO-independent mechanisms. The failure of CC-chemokines to affect VLA-4 and Mac-1 expression suggests that changes in integrin function (avidity or affinity) are rather involved in the enhanced adhesion. © 2008 Lintomen et al; licensee BioMed Central Ltd.

Scientific article in vitro evaluation of the influence of air abrasion on detection of occlusal caries lesions in primary teeth

The purpose of this study was to assess the influence of cleaning pits and fissures with an aluminum oxide air abrasion system on the detection of occlusal caries in primary teeth using laser fluorescence (LF) and visual examination. Methods: The sample comprised 65 pit and fissure sites on extracted primary teeth suspected to be carious. The sites were submitted to 2 visual examinations (examiner JAR) and 2 LF readings (examiner TMV). Next, the occlusal surfaces were air-abraded and re-examined thereafter using both methods. The teeth were sectioned, and the histological analysis of the sites with a stereoscopic magnifying lens at X32 magnifi cation was used as the gold standard. Results: Cohen's kappa statistic for LF and visual examination were, respectively, 0.282/0.884 before and 0.896/0.905 after air abrasion. LF showed a sensitivity of 0.28 increasing to 0.49 and a specifi city of 0.50 increasing to 0.92. Visual examination showed sensitivity of 0.78 and specifi city of 0.73. Both increased after air abrasion. Conclusion: The findings suggest that cleaning pits and fissures with aluminum oxide air abrasion increased the accuracy of LF and visual examination for detection of occlusal caries in primary teeth.

In vitro evaluation of the effect of dietary acids and toothbrushing on human root dentin permeability

OBJECTIVES: The purpose of this in vitro study was to quantify the alterations in human root dentin permeability after exposure to dietary acids and to evaluate the effect of toothbrushing after acid application. METHOD AND MATERIALS: Extracted human third molars had their crowns sectioned above the CEJ, pulp tissue removed, and cervical root dentin exposed using a high-speed bur (approximately 1 mm in depth of substance loss). From each root fragment, one specimen was prepared. A total of 25 specimens were used and distributed randomly into five groups. The specimens were attached to a hydraulic pressure apparatus to evaluate the alterations of root dentin permeability after exposure to different acids. Dentin permeability was measured after the following sequential steps: (1) treatment with EDTA for 3 minutes to obtain the maximum permeability; (2) root planing to create a smear layer; (3) exposure to different acidic substances for 5 minutes (vinegar, cola drink, lemon juice, white wine, and orange juice); and (4) brushing for 3 minutes. RESULTS: All acidic substances increased dentin permeability after root planing. Lemon juice produced higher values for permeability when compared to the other substances (P = .009); moreover, orange juice showed similar results (P < .02) except when compared to vinegar (P = .12). Brushing right after acid exposure significantly reduced dentin permeability except in the vinegar group (P = .07). CONCLUSION: Under the experimental conditions, dietary acids increased root dentin permeability, and immediate brushing reduced permeability levels.

Chromatin modifying agents in the in vitro production of bovine embryos

The low efficiency observed in cloning by nuclear transfer is related to an aberrant gene expression following errors in epigenetic reprogramming. Recent studies have focused on further understanding of the modifications that take place in the chromatin of embryos during the preimplantation period, through the use of chromatin modifying agents. The goal of these studies is to identify the factors involved in nuclear reprogramming and to adjust in vitro manipulations in order to better mimic in vivo conditions. Therefore, proper knowledge of epigenetic reprogramming is necessary to prevent possible epigenetic errors and to improve efficiency and the use of in vitro fertilization and cloning technologies in cattle and other species. Copyright © 2011 Fabio Morato Monteiro et al.

In vitro susceptibility of oral Candida albicans strains to different pH levels and calcium hydroxide saturated aqueous solution

Candida albicans is present in the oral cavity and in the whole digestive tract of humans and other animals, being frequently related to endodontic treatment failure. The present study determined the incidence of C. albicans in the oral cavity and the susceptibility of isolates to different pH values and saturated calcium hydroxide aqueous solution at pH 12.5. Sixty-five patients attending the Endodontic Clinic at the Sagrado Coração University participated in the study. The collected samples were cultivated in selective media for C. albicans and the isolates were tested in terms of resistance to both alkaline pH and saturated aqueous solution of calcium hydroxide. In relation to time variables, yeast viability was assessed by the Sabouraud's agar culture and fluorescein diacetate and ethidium bromide fluorescent staining method. Results from the different pHs and experimental times, including those from different techniques measuring fungal viability, were compared using the chi-square and Fisher's exact tests (α=0.05). The yeasts became completely inviable after 48 h of contact with the calcium hydroxide solution. On the other hand, when exposed to the alkaline culture broth, the yeasts were found to be viable at pHs 9.5 and 10.5 for up to 7 days. In conclusion, C. albicans can only be completely inhibited by direct contact with saturated calcium hydroxide aqueous solution after 48 h of exposure.

Seed and seedling surface-sterilization for in vitro culture of tillandsia gardneri (Bromeliaceae)

Tillandsia gardneri is a bromeliad with ornamental value and a wide geographical distribution over Brazil. However, due to habitat loss and illegal overcollection in the wild it is included as a vulnerable species in the official list of endangered plants of the State of Rio Grande do Sul, Brazil. The development of a protocol for T. gardneri seed propagation in vitro may be useful for reintroducing plants in their natural habitats, and for germplasm conservation. A difficult problem encountered during the establishment of an in vitro culture is explants disinfection, especially when working with endangered species, from which explant availability is restricted. Thus, the establishment of a sterilization protocol is crucial for the initiation and success of a micropropagation system for T. gardneri. The objective of this study was to evaluate the effect of sodium hypochlorite concentration and exposure time in seed and seedling surface disinfection, tissue sensitivity and development. Sodium hypochlorite solutions (10 or 20%/5, 10 or 15 min; 25%/5 or 10 min; and 50%/5 min) were effective in eliminating seed superficial contaminants. There was no significant difference among the effective sterilization treatments in relation to seed germination (%), and seedling length and number of leaves, after 120 days in vitro. Also, no damage to seed and seedling tissues were observed. Surface sterilization of seedlings, for initiation of an in vitro culture, required higher concentrations of sodium hypochlorite (25%/15 min; 20 or 50%/5, 10 or 15 min; and 40%/5 and 10 min) for controlling fungal and yeast contamination, compared to seed sterilization. No significant differences among these treatments were found in relation to seedling length and number of leaves, after 60 days in vitro.

Effects of ascorbic acid on in vitro culture of bovine preantral follicles

The objective of this study was to evaluate the effects of adding ascorbic acid to the media for in vitro culture of cattle ovarian fragments and to determine their effects on growth activation and viability of early-stage follicles. The ovarian cortex was divided into small fragments; one fragment was immediately fixed (control) and the other fragments were cultured in minimum essential medium (MEM) supplemented or not with various doses of ascorbic acid. Ovarian tissue was processed for histology, transmission electron microscopy (TEM) and immunohistochemical demonstration of proliferating cell nuclear antigen (PCNA). Compared with control fragments, the percentage of primordial follicles was reduced (p < 0.05) and the percentage of growing follicles had increased (p < 0.05) in cultured cortical fragments, independent of the tested medium or incubation time. Furthermore, compared with control tissue, culture of ovarian cortex for 8 days reduced the percentages of healthy, viable follicles (p < 0.05), but not when cultures were supplemented with 25, 50 or 100 μg/ml of ascorbic acid. Ultrastructural and immunohistochemical analysis of 8 day cultured ovarian cortical fragments, however, showed the integrity and viability of follicles only when fragments were cultured in presence of 50 μg/ml of ascorbic acid. In conclusion, this study demonstrated that addition of ascorbic acid to MEM at a concentration of 50 μg/ml not only stimulates the activation of 8 day in vitro cultured cattle primordial follicles and subsequent growth of activated follicles, but also safeguards the viability of these early-stage follicles. © 2012 Copyright Cambridge University Press.

In vitro assessment of dentin erosion after immersion in acidic beverages: Surface profile analysis and energy-dispersive X-ray fluorescence spectrometry study

The aim of this study was to investigate the effects of some acidic drinks on dentin erosion, using methods of surface profile (SP) analysis and energy-dispersive X-ray fluorescence spectrometry (EDXRF). One hundred standardized dentin slabs obtained from bovine incisor roots were used. Dentin slabs measuring 5x5 mm were ground flat, polished and half of each specimen surface was protected with nail polish. For 60 min, the dentin surfaces were immersed in 50 mL of 5 different drinks (Gatorade®, Del Valle Mais orange juice®, Coca-Cola®, Red Bull® and white wine), 20 blocks in each drink. The pH of each beverage was measured. After the erosive challenge, the nail polish was removed and SP was analyzed. The mineral concentration of dentin surfaces was determined by means of EDXRF. Data were analyzed statistically by ANOVA and Tukey's test (α=0.05). SP analysis showed that Red-Bull had the highest erosive potential (p<0.05). EDXRF results exhibited a decrease in phosphate in the groups immersed in Red-Bull, orange juice and white wine (p<0.05), and no significant difference in calcium content between the reference surface and eroded surface. In conclusion, this study demonstrated that all studied beverages promoted erosion on root dentin and Red Bull had the highest erosive potential. There was no correlation between pH of beverages and their erosive potential and only the P content changed after erosive challenge.