Analysis of the effects of thermal cycling on the microtensile shear bond strength of a self-etching and a conventional pit and fissure sealants to dental enamel

Objective: To analyze the effects of thermal cycling on the microtensile shear bond strength of a self-etching and a conventional pit and fissure sealants to dental enamel. Material and Method: Twenty-four healthy human molars extracted for orthodontic reasons, were sectioned in the mesio-distal direction and divided into two groups (n=24) according to the sealant to be applied: GI - conventional sealant Climpro (3M/ESPE) and GII - self-etching sealant Enamel Loc (Premier Dental). The sealants were applied on flattened enamel in matrixes 1 mm in diameter, in accordance with the manufacturers' recommendations. The specimens were stored in distilled water at 37°C for 24 hours. After this, half the samples of both groups were submitted to 500 thermal cycles in 30s baths at temperatures between 5 and 55°C. Forty-eight hours after the samples were made, the microtensile shear test was performed in an Instron 4411 test machine, with a stainless steel wire with a cylindrical cross section of 0.2mm in diameter at a constant speed of 0.5mm/s. The bond strength values were submitted to ANOVA for 2 factors and the fracture patterns were examined under an optical microscope at 65X magnification. Results: Thermal cycling did not influence the bond strength of the two sealants. The conventional sealant Climpro presented a statistically higher microtensile shear bond strength (11.72MPa, 11.34MPa with and without cycling, respectively) than the self-etching sealant Enamel Loc (5.92MPa, 5.02MPa with and without cycling, respectively). Fracture pattern analysis showed the occurrence of 100% of adhesive failures for Enamel Loc, while the conventional sealant Climpro presented 95% of adhesive failures and 5% of mixed failures. Conclusion: The conventional sealant presented higher microtensile shear bond strength to dental enamel in comparison with the self-etching sealant. Thermal cycling did not affect the bond strength of the sealants used in this study. © 2011 Nova Science Publishers, Inc.

Prevalence and microscopic features of enamel pearls from permanent human molars

Enamel pearls are ectopic structures observed mainly on the roots of permanent teeth and could be related to periodontal diseases. Aim: To evaluate the prevalence of enamel pearls in extracted human molars and characterize their structures using light and scanning electron microscopy. Methods: The study comprised 2,201 extracted human permanent molars. The teeth were analyzed and classified according to morphological features. The presence, location and shape of enamel pearls were investigated. Fifteen human molars with enamel pearls were embedded in acrylic resin and observed by light microscopy. Results: Seventy-one enamel pearls were identified on third molar root. Microscopically, most pearls were composed of prismatic irregular enamel and normal dentin. The dentinoenamel junction presented an irregular course. The number of dentinal tubules was normal and they presented curvature to continue within the root dentin of the carrier tooth. Dentinal tubules below the enamel pearls were closer to each other. Conclusions: Scanning electron microscopic analysis revealed that the enamel pearls were similar to coronal enamel.

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.

Fracture resistance of bovine incisors restored with different glass fiber posts: Effect of the diameter of fiber post

Aim: Compare the effect of three post designs on the fracture resistance and failure modes of composite core-fiber post-crownless tooth sets. Materials and Methods: Ninety bovine incisors were selected and divided into nine groups of 10 specimens. The teeth were assigned to three groups based on the post design: Cylindrical, tapered, and double-tapered. Each group was subdivided into three subgroups in accordance with the diameter of the post: Small (No.1), medium (No.2), and large (No.3). The Panavia F system was used for post cementation. The specimens were mounted in acrylic resin blocks with a layer of silicone rubber covering the roots. A universal testing machine compressively loaded the specimens from the palatal side at a crosshead speed of 1 mm/min and at an angle of 135I to the long axis of the teeth, until failure occurred. The failure mode was determined by a stereomicroscope inspection of all the specimens. Data were analyzed by one-way ANOVA and the Tukey test (P < 0.05). Results: The fracture resistance was affected by the type of post (P < 0.0001). A narrower diameter for all of the post systems allowed for higher resistance. The main failure mode in the large cylindrical group was catastrophic fractures, while the main failures in the other eight groups were favorable. Conclusion: Narrower diameter posts showed higher fracture resistance. The dominant failure pattern was repairable fracture, except for those with large cylindrical groups.

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.

Dispersed hydroxyapatite bioglass 45S5 composites: Comparative evaluation of the use of bovine bone and synthetic hydroxyapatite

The bovine bone and sintetic hydroxyapatite (HA) bioceramics are reference materials to employment as a bone substitute, however, their slow rate of degradation and its low rate of bioactivity index (Ib) are presented as limiting factors for application as bone graft. In contrast, the bioglass is a resorbable and osteoinductive material. the present work objective the development of composites of dispersed bovine bone or sintetic HA in silicate-phosphate bioglass, seeking to obtain a biomaterial with properties suitable for application as bone grafts. The composites were prepared by mixing between the powder components followed by sintering for 1h. Were used HA and bioglass (45S5) with particle size <240μm. The tested proportions of HA/45S5 were 20/80, 30/70 and 40/60 (wt%). The composites characterization was made employing scanning electron microscopy, Infra-Red Spectrometry and hydrolytic resistance test. The test results indicate the potential use of the materials developed for applications such as bone graft. © (2012) Trans Tech Publications, Switzerland.

Effect of fluoride solutions on the shear bond strength of orthodontic brackets

The aim of this in vitro study was to evaluate the shear bond strength of brackets after pre-treatment with different fluoride solutions. This study used 48 freshly extracted sound bovine incisors that were randomly assigned to 4 experimental groups (n=12). CG: (control) without treatment; NF: 4 min application of neutral fluoride; APF: application of 1.23% acidulated phosphate fluoride (APF) for 4 min; and SFV: application of 5% sodium fluoride varnish for 6 h. For each group, after surface treatment, prophylaxis of enamel and bracket bonding with Transbond XT composite resin (3M) were performed following the manufacturer's specifications. The shear bond strength was performed with a universal testing machine 24 h after fixing the brackets. The tooth surfaces were analyzed to verify the adhesive remnant index (ARI). Data were analyzed statistically by ANOVA and Tukey's test (α=0.05). There was statistically significant difference among the groups (p<0.0001). CG and NF groups presented significantly higher bond strength than APF and SFV. There was no significant difference between CG and NF or between APF and SFV (p>0.05). The analysis of ARI scores revealed that most failures occurred at the enamel-resin interface. It may be concluded that the pre-treatment of enamel with 1.23% APF and 5% SFV prior to fixing orthodontic brackets reduces shear bond strength values.

Efficacy and cytotoxicity of a bleaching gel after short application times on dental enamel

Objectives: This study aimed to evaluate and correlate the efficacy and cytotoxicity of a 35 % hydrogen peroxide (HP) bleaching gel after different application times on dental enamel. Materials and methods: Enamel/dentin disks in artificial pulp chambers were placed in wells containing culture medium. The following groups were formed: G1, control (no bleaching); G2 and G3, three or one 15-min bleaching applications, respectively; and G4 and G5, three or one 5-min bleaching applications, respectively. Extracts (culture medium with bleaching gel components) were applied for 60 min on cultured odontoblast-like MDPC-23 cells. Cell metabolism (methyl tetrazolium assay) (Kruskal-Wallis/Mann-Whitney; α = 5 %) and cell morphology (scanning electron microscopy) were analyzed immediately after the bleaching procedures and the trans-enamel and trans-dentinal HP diffusion quantified (one-way analysis of variance/Tukey's test; α = 5 %). The alkaline phosphatase (ALP) activity was evaluated 24 h after the contact time of the extracts with the cells (Kruskal-Wallis/Mann-Whitney; α = 5 %). Tooth color was analyzed before and 24 h after bleaching using a spectrophotometer according to the Commission Internationale de l'Eclairage L*a*b* system (Kruskal-Wallis/Mann-Whitney; α = 0.05). Results: Significant difference (p < 0.05) in cell metabolism occurred only between G1 (control, 100 %) and G2 (60.6 %). A significant decrease (p < 0.05) in ALP activity was observed between G2, G3, and G4 in comparison with G1. Alterations on cell morphology were observed in all bleached groups. The highest values of HP diffusion and color alterations were observed for G2, with significant difference among all experimental groups (p < 0.05). G3 and G4 presented intermediate color change and HP diffusion values with no statistically significant differences between them (p > 0.05). The lowest amount of HP diffusion was observed in G5 (p < 0.05), which also exhibited no significant color alteration compared to the control group (p > 0.05). Conclusions: HP diffusion through dental tissues and its cytotoxic effects were proportional to the contact time of the bleaching gel with enamel. However, shorter bleaching times reduced bleaching efficacy. Clinical relevance: Shortening the in-office tooth bleaching time could be an alternative to minimize the cytotoxic effects of this clinical procedure to pulp tissue. However, the reduced time of bleaching agent application on enamel may not provide adequate esthetic outcome. © 2012 Springer-Verlag Berlin Heidelberg.

HDAC inhibition decreases XIST expression on female IVP bovine blastocysts

During initial development, both X chromosomes are active in females, and one of them must be silenced at the appropriate time in order to dosage compensate their gene expression levels to male counterparts. Silencing involves epigenetic mechanisms, including histone deacetylation. Major X chromosome inactivation (XCI) in bovine occurs between hatching and implantation, although in vitro culture conditions might disrupt the silencing process, increasing or decreasing X-linked gene expression. In this study, we aimed to address the roles of histone deacetylase inhibition by trichostatin A (TSA) on female preimplantation development.We tested the hypothesis that by enhancing histone acetylation, TSA would increase the percentage of embryos achieving 16-cell stage, reducing percentage of embryos blocked at 8-cell stage, and interfere with XCI in IVF embryos. We noticed that after TSA treatment, acetylation levels in individual blastomeres of 8-16 cell embryos were increased twofold on treated embryos, and the samewas detected for blastocysts. Changes among blastomere levels within the same embryo were diminished on TSA group, as low-acetylated blastomeres were no longer detected. The percentage of embryos that reached the 5th cleavage cycle 118 h after IVF, analyzed by Hoechst staining, remained unaltered after TSA treatment. Then, we assessed XIST and G6PD expression in individual female bovine blastocysts by quantitative real-time PCR. Even though G6PD expression remained unaltered after TSA exposure, XIST expression was eightfold decreased, and we also detected a major decrease in the percentage of blastocysts expressing detectable XIST levels after TSA treatment. Based on these results, we conclude that HDAC is involved on XCI process in bovine embryos, and its inhibition might delay X chromosome silencing and attenuate aberrant XIST expression described for IVF embryos. © 2013 Society for Reproduction and Fertility.

Efficacy of mouth rinses and toothpaste on tooth whitening.

People increasingly desire tooth whitening. Considering the wide range of whitening products on the market, this study evaluated the efficacy of whitening toothpastes and mouth rinses compared with the 10% carbamide peroxide (CP) whitening gel. We obtained 120 cylindrical specimens from bovine teeth, which were darkened for 24 hours in a coffee solution. The color measurement was performed by a spectrophotometer using the CIE L*a*b* system, and specimens were divided into six groups according to the use of the following agents: group 1, conventional fluoridated toothpaste; group 2, Close Up White Now; group 3, Listerine Whitening; group 4, Colgate Plax Whitening; group 5, experimental mouth rinse with Plasdone; and group 6, 10% CP Whiteness Perfect. After the simulation of 12 weeks of treatment for groups 1 to 5 and 14 days of treatment for group 6, the specimens were subjected to a new color reading. Data were subjected to one-way analysis of variance (α=0.05), which showed significant differences among groups after 12 weeks for ΔE (p=0.001). Results of the Tukey test revealed that groups 3, 4, and 6 presented significantly higher color alteration than groups 1, 2, and 5. The whitening toothpaste Close Up White Now and the experimental mouth rinse with Plasdone showed similar color alteration as conventional toothpaste after a 12-week treatment simulation. These groups presented significantly lower color alteration compared with whitening mouth rinses Listerine and Colgate Plax Whitening, which showed similar results to those observed after 14 days of bleaching with 10% CP treatment.

Autogenous bone combined with anorganic bovine bone for maxillary sinus augmentation: Analysis of the osteogenic potential of cells derived from the donor and the grafted sites

Objectives: This study aimed to comparatively evaluate the in vitro osteogenic potential of cells obtained from the mandibular ramus (MR, autogenous bone donor site) and from the maxillary sinus (MS) bone grafted with a mixture of anorganic bovine bone (ABB) and MR prior to titanium implant placement (MS, grafted implant site). Material and methods: Cells were obtained from three patients subjected to MS floor augmentation with a 1: 1 mixture of ABB (GenOx Inorg®) and MR. At the time of the sinus lift procedure and after 8 months, prior to implant placement, bone fragments were taken from MR and MS, respectively, and subjected to trypsin-collagenase digestion for primary cell culturing. Subcultured cells were grown under osteogenic condition for up to 21 days and assayed for proliferation/viability, osteoblast marker mRNA levels, alkaline phosphatase (ALP) activity and calcium content/Alizarin red staining. ALP activity was also determined in primary explant cultures exposed to GenOx Inorg® (1: 1 with MR) for 7 days. Data were compared using either the Mann-Whitney U-test or the Kruskal-Wallis test. Results: MS cultures exhibited a significantly lower osteogenic potential compared with MR cultures, with a progressive increase in cell proliferation together with a decrease in osteoblast markers, reduced ALP activity and calcium content. Exposure of MR-derived primary cultures to GenOx Inorg® inhibited ALP activity. Conclusion: These results suggest that the use of GenOx Inorg® in combination with MR fragments for MS floor augmentation inhibits the osteoblast cell differentiation at the implant site in the long term. © 2013 John Wiley & Sons A/S.

Supplementation with the histone deacetylase inhibitor trichostatin A during in vitro culture of bovine embryos

Summary Trichostatin A (TSA) is a histone deacetylase inhibitor that induces histone hyperacetylation and increases gene expression levels. The aim of the present study was to establish a suitable condition for the use of TSA in in vitro cultures of bovine embryos, and to determine whether TSA would increase blastocyst rates by improvement of chromatin remodelling during embryonic genome activation and by increasing the expression of crucial genes during early development. To test this hypothesis, 8-cell embryos were exposed to four concentrations of TSA for different periods of time to establish adequate protocols. In a second experiment, three experimental groups were selected for the evaluation of embryo quality based on the following parameters: apoptosis, total cell number and blastocyst hatching. TSA promoted embryonic arrest and degeneration at concentrations of 15, 25 and 50 nM. All treated groups presented lower blastocyst rates. Exposure of embryos to 5 nM for 144 h and to 15 nM for 48 h decreased blastocyst hatching. However, the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay (TUNEL) assay revealed similar apoptosis rates and total cell numbers in all groups studied. Although, in the present study, TSA treatment did not improve the parameters studied, the results provided background information on TSA supplementation during in vitro culture of bovine embryos and showed that embryo quality was apparently not affected, despite a decrease in blastocyst rate after exposure to TSA. © Cambridge University Press 2011.

Canonical WNT signaling regulates development of bovine embryos to the blastocyst stage

Objectives were to evaluate the role of canonical WNT signaling in development of the preimplantation embryo. Signaling was activated with 2-Amino-4-(3,4-(methylenedioxy)benzylamino)-6-(3-methoxyphenyl)pyrimidine (AMBMP) and inhibited with Dickkopf-related protein 1 (DKK1). Treatment of bovine embryos with AMBMP at day 5 after insemination decreased development to the blastocyst stage at day 7 and reduced numbers of trophectoderm and inner cell mass cells. At high concentrations, AMBMP caused disorganization of the inner cell mass. DKK1 blocked actions of AMBMP but did not affect development in the absence of AMBMP. Examination of gene expression in day 6 morulae by microarray revealed expression of 16 WNT genes and other genes involved in WNT signaling; differences in relative expression were confirmed by PCR for 7 genes. In conclusion, the preimplantation embryo possesses a functional WNT signaling system and activation of the canonical pathway can inhibit embryonic development.

Effect of fluoride-treated enamel on indirect cytotoxicity of a16% carbamide peroxide bleaching gel to pulp cells

The aim of this study was to evaluate the possibility of fluoride solutions applied to enamel to protect pulp cells against the trans-enamel and transdentinal cytotoxicity of a 16% carbamide peroxide (CP) bleaching gel. The CP gel was applied to enamel/ dentin discs adapted to artificial pulp chambers (8 h/day) during 1, 7 or 14 days, followed by fluoride (0.05% or 0.2%) application for 1 min. The extracts (culture medium in contact with dentin) were applied to MDPC-23 cells for 1 h, and cell metabolism (MTT assay), alkaline phosphatase (ALP) activity and cell membrane damage (flow cytometry) were analyzed. Knoop microhardness of enamel was also evaluated. Data were analyzed statistically by ANOVA and Kruskal-Wallis tests (a=0.05). For the MTT assay and ALP activity, significant reductions between the control and the bleached groups were observed (p<0.05). No statistically significant difference occurred among bleached groups (p>0.05), regardless of fluoride application or treatment days. Flow cytometry analysis demonstrated 30% of cell membrane damage in all bleached groups. After 14 days of treatment, the fluoride-treated enamel presented significantly higher microhardness values than the bleached-only group (p<0.05). It was concluded that, regardless of the increase in enamel hardness due to the application of fluoride solutions, the treated enamel surface did not prevent the toxic effects caused by the 16% CP gel to odontoblast-like cells.