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ABSTRACT: We have recently reported that the signal of pulp injury induces both neuronal and glial cell activation in the contralateral thalamus in rats, although the mechanisms of the glial cell/neuronal interaction remain unclear. This study was undertaken to test our hypothesis that p38 mitogen-activated protein kinase (MAPK) signaling pathways are involved in the pulp injury-induced glial cell/neuronal interaction in the thalamus.
A local anesthetic (lidocaine with epinephrine) or saline (control) was injected into the tissue surrounding the left mandibular first molar of Wistar rats. The tooth was then pulp-exposed, and the cavity was sealed with flowable composite. After 0 (normal pulp with local anesthetic or saline pretreatment), 24, and 72 hours, the contralateral side of thalamus was retrieved and subjected to immunohistochemistry for phospho-p38 MAPK and glial fibrillary acidic protein and real-time polymerase chain reaction analysis of p38-MAPK family (MAPK 13 and MAPK 14) mRNAs.
The area immunopositive to phospho-p38 MAPK increased until 72 hours after pulp exposure in both local anesthetic-pretreated and saline-pretreated animals, but the rate of increase was lower in the local anesthetic-pretreated animals. The density of glial fibrillary acidic protein-expressing astrocytes showed a significant increase only in the saline-pretreated animals. Expression levels of MAPK 13 and MAPK 14 mRNAs increased at 24 hours and still higher at 72 hours in the saline-pretreated animals. Notably, MAPK 13 and MAPK 14 mRNA levels at 24 and 72 hours in the local anesthetic-pretreated animals showed significantly lower levels than those in the saline-pretreated animals.
It was concluded that pulp injury-induced up-regulation of MAPK 13, MAPK 14, and phospho-p38 MAPK in the thalamus was suppressed by the local anesthetic pretreatment, suggesting the involvement of p38 MAPK signaling pathways in the glial cell-neuronal interaction induced by pulpal nociception.
Journal of endodontics 04/2013; 39(4):488-92. · 2.95 Impact Factor
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ABSTRACT: The aim of this study was to investigate coronal leakage after obturation with mineral trioxide aggregate (MTA), resin-based sealer, and silicon-based sealer for open apical foramina and to evaluate pathway of leakage. Twenty-eight maxillary premolars were used, and instrumented to ISO size #80. Teeth were randomly divided into four groups as follows: Group A filled with MTA, Group B with gutta-percha and resin-based sealer, Group C with polymer-based material and resin-based sealer, and Group D with gutta-percha and silicon-based sealer. All samples were evaluated for coronal leakage with methylene blue solution and spectrophotometry. After leakage testing, samples were cut, and sections were observed. Dye leakage of Group A was significantly lowest among all groups at 15 days and 30 days. Defects which induced coronal leakage in resin-based sealer were observed at 7 mm from the apex. Coronal leakage after obturation with MTA for open apical foramina was significantly lower than resin-based sealer and silicon-based sealer.
Dental Materials Journal 01/2013; 32(1):130-137. · 1.14 Impact Factor
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ABSTRACT: Objective: The purpose of this study was to compare optical coherence tomography (OCT) with the existing technologies, to assess its accuracy and utility in detecting vertical root fractures of extracted human teeth. Background data: The detection of root fractures in teeth that have undergone root canal treatment is challenging because of the great difficulty in differentiating these fractures from morphologic or radiographic anomalies. OCT methods are based on depth-resolved optical reflectivity and have been developed to reduce the invasiveness and radiation exposure inherent to other techniques. Methods: Twelve extracted human mandibular teeth (totaling 25 roots) that were free of caries, calculus, and root treatment were used, and assessed by microfocus computed tomography, the current gold standard for fracture detection. The ability of appropriately trained observers to detect root fractures using visual, microscopic, and swept-source OCT (SS-OCT) techniques were compared. micro-CT and SS-OCT produce three-dimensional images of the tooth from which to diagnose fractures, but CT scanning involves radiation exposure that is not required in SS-OCT. Results: Seventeen of the 25 roots were found to have fractures by microfocus CT. These findings were replicated by SS-OCT, which revealed fractures exhibiting identical origin, size, and angulation within the root. We found that SS-OCT gave results compatible to the gold standard technique, and that SS-OCT and microscopy were more effective for identifying root fractures than was visual observation alone. Conclusions: SS-OCT may represent a novel, noninvasive, noncontact and nonexposure alternative to the conventional methods used for assessing root fractures in teeth.
Photomedicine and laser surgery 12/2012; · 1.76 Impact Factor
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ABSTRACT: Hertwig's epithelial root sheath (HERS), a bilayered epithelial cell sheath located at the cervical loop of the enamel organ in a developing tooth, is at the forefront of root formation. However, little is known about the exact mechanisms that regulate the development of HERS. The neuropeptide vasoactive intestinal peptide (VIP) is involved in the development of various tissues and cells. In this study, we investigated the roles of VIP in HERS development. VIP-immunoreactive nerve fibers were found in the dental pulp and around the root apex of the tooth, while the expression of VIP receptor 1 (VPAC1) was observed in HERS. The expression level of VPAC1 correlated with the development of HERS and was elevated at postnatal days 14 and 21. Using ex vivo cultures of neonatal tooth germs, VIP enhanced the elongation and proliferation of HERS. In vitro, VIP also promoted the proliferation of cells from the HERS-derived cell line, HERS01a cells, and upregulated the mRNA expression of cytokeratin 14 and vimentin (typical molecular markers of HERS) in these cells. These results suggest that VIP may be an essential factor for HERS development.
Congenital Anomalies 09/2012; 52(3):162-7.
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ABSTRACT: OBJECTIVES: The objectives of the present study were to quantitatively evaluate chemical permeability through human enamel/dentine using conductometry and to clarify if alternating current (AC) iontophoresis facilitates such permeability. MATERIALS AND METHODS: Electrical impedance of different concentrations of lidocaine hydrochloride was measured using a bipolar platinum impedance probe. A quadratic curve closely fitted to the response functions between conductance and lidocaine hydrochloride. For analysis of the passage of lidocaine hydrochloride through human enamel/dentine, eight premolars that were extracted for orthodontic treatment were sectioned at the cemento-enamel junction. The tooth crowns were held between two chambers with a double O-ring. The enamel-side chamber was filled with lidocaine hydrochloride, and the pulp-side chamber was filled with extrapure water. Two platinum plate electrodes were set at the end of each chamber to pass alternating current. A simulated interstitial pulp pressure was applied to the pulp-side chamber. The change in the concentration of lidocaine hydrochloride in the pulp-side chamber was measured every 2min using a platinum recording probe positioned at the centre of the pulp-side chamber. Passive entry without iontophoresis was used as a control. RESULTS: The level of lidocaine hydrochloride that passed through enamel/dentine against the dentinal fluid flow increased with time. Electrical conductance (G, mho) correlated closely to the concentration (x, mmol/L) of lidocaine hydrochloride (G=2.16x(2)+0.0289x+0.000376, r(2)=0.999). CONCLUSIONS: Lidocaine hydrochloride can pass through enamel/dentine. Conductometry showed that the level of lidocaine hydrochloride that passed through enamel/dentine was increased by AC iontophoresis.
Archives of oral biology 08/2012; · 1.65 Impact Factor
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ABSTRACT: The purpose of the present study was to investigate the degree of Er:YAG laser irradiation at the apical area in vitro.
Since the laser was developed, advancement of laser treatment has been seen in various fields. However, few reports exist on shaping of the root canal using Er:YAG laser irradiation.
Six single-rooted human teeth were used. The working length of root canals was set at 6.5 mm, and they were enlarged to apical file size #25. An Er:YAG laser and cone-shaped irradiation tips (R135T and R200T) were used. Laser irradiation conditions were 30 m J, 20 pps, and water flow of 5 mL/min. Samples were irradiated three times for 10 sec each using each tip. To evaluate the cutting degree of horizontal area of the root canal, the laser-irradiated surfaces were observed using microfocus X-ray computed tomographic photography before and after every irradiation. The samples were observed under a scanning electron microscope. Measurement of pixels in an area was performed by image-editing software (Adobe Photoshop 7.0). Statistical analysis was performed using StatView (version 5.0). One-way ANOVA and the Tukey-Kramer tests were used; p<0.05 indicated statistical significance.
When root canals were irradiated with R200T for 10 sec (p<0.05), a large amount of evaporation (0.12 ± 1.07 mm(2)) was acquired in their cut area compared with the other irradiation conditions. In scanning electron microscopic observation, there was no smear layer and the dentinal tubules were open.
When the distance between the tip and root dentin was adjacent, the shaping of root dentin by Er:YAG laser irradiation was definitely observed.
Photomedicine and laser surgery 06/2012; 30(7):367-73. · 1.76 Impact Factor
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ABSTRACT: In conventional whole-tooth culture systems, limitation exists regarding maintenance of the vitality of the dental pulp, because this tissue is encased in rigid dentin walls that hinder nutrition supply. We here report a whole tooth-in-jaw-bone culture system of rat mandibular first molars, where transcardiac perfusion with culture medium was carried out before placement of the jaw bone into culture medium, aiming to facilitate longer time preservation of the dental pulp tissue. Following 7 days of culture, the pulp tissues were analyzed by histology and immunohistochemistry to ED2 (antiresident macrophage). ED2-positive macrophages were also analyzed for their Class II MHC, interleukin-6 (IL-6), and p53 mRNA expression levels by means of immune-laser capture microdissection (immune-LCM). Dentin sialophosphoprotein (DSPP) mRNA expression in odontobalstic layer was also examined by LCM. Teeth cultured following saline-perfusion and nonperfusion served as cultured controls. Normal teeth also served as noncultured controls. Histological examination demonstrated that the structure of the pulp tissue was well preserved in the medium-perfused explants in contrast to the cultured control groups. The Class II MHC, IL-6, and p53 mRNA expression levels of ED2-positive cells and DSPP expression levels of odontoblastic layer tissues in the pulp of medium-perfused explants were not significantly different from those in the noncultured normal teeth. In conclusion, the structural integrity and mRNA expression in the pulp were maintained at the in vivo level in the ex vivo whole tooth-in-jaw-bone culture system. The system may lay the foundation for studies aiming at defining further histological and molecular mechanism of the pulp. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.
Microscopy Research and Technique 05/2012; 75(10):1341-7. · 1.79 Impact Factor
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ABSTRACT: The dispersion of the lifetime of NiTi instruments, and their deflecting load (DL) changes during cyclic fatigue were investigated. A total of 120 ProFile NiTi rotary instruments were tested using a specially designed cyclic fatigue testing apparatus with three pins. Using these pins, the instrument was bent and rotated at 300 rpm to fracture. DL was recorded using a load cell attached to the central pin. For each sample, the working time was converted to number of cycles to fracture (NCF) and the mean DL (DL(m)) was calculated. The averages of NCF and DL(m) of 120 samples were 584.3±180.5 cycles and 6.44±0.91 N, respectively. All samples showed a sequential decrease in DL during rotation. Based on the present study, it is impossible to estimate the lifetime of a NiTi instrument from NCF. Thus, the change in DL could be an alternative criterion to determine the remaining lifetime.
Dental Materials Journal 05/2012; 31(3):389-93. · 1.14 Impact Factor
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ABSTRACT: The purpose of this in vitro study was to evaluate the effect of surface modifications induced by erbium (Er):YAG and neodymium (Nd):YAG laser irradiation on cell adhesion by comparing it to that of conventional methods for surface preparation after root-end resection.
Many studies have been seeking a favorable method to produce a resected root end with optimal conditions for cell response. However, little improvement has been achieved. This study evaluated the biocompatibilities of resected root surfaces after Er:YAG or Nd:YAG laser irradiation on initial cell adhesion.
Dentin disks were divided into three groups. Group A was left untreated, Group B was treated with Er:YAG laser irradiation (60 mJ/pulse, 10 pps, 60 sec), and Group C with Nd:YAG laser irradiation (60 mJ/pulse, 10 pps, 60 sec). After laser irradiation, the dentin disks were incubated with NIH/3T3 fibroblasts cultured in Dulbecco's modified Eagle's medium. A morphological analysis of the dentin surface and cell adhesion was observed under a scanning electron microscope. Surface roughness was measured using a confocal laser scanning microscope. The statistical analysis was undertaken using ANOVA at a level of significance of 5% (p<0.05).
Morphological analysis and roughness measurement showed that dentin surfaces treated with Er:YAG laser irradiation were rougher than those in Groups A and C. Group B (Er:YAG) exhibited the greatest number of attached cells among all groups after 12 and 24 h.
Morphological alteration induced by Er:YAG laser irradiation showed a favorable effect on the attachment of fibroblasts to dentin surfaces.
Photomedicine and laser surgery 11/2011; 30(2):63-70. · 1.76 Impact Factor
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ABSTRACT: In normal dental pulp, a considerable number of resident macrophages are distributed. This study was designed to analyze the expression levels of genes associated with differentiation and function of resident macrophages in rat molar pulps stimulated with lipopolysaccharide (LPS).
Mandibular first molars of 7-week-old male Wistar rats were used. After transcardiac perfusion with a culture medium to preserve tissue integrity, pulpotomy and LPS application were carried out on the experimental teeth, and then dissected mandibles were subjected to whole-tooth culture for 3 days. Normal teeth and pulpotomized teeth without LPS served as controls. The specimens were then immunostained for ED1 (CD68, a general macrophage marker) and ED2 (CD163, a resident macrophage marker). Real-time polymerase chain reaction for Toll-like receptor 4 (TLR4), CD14, chemokine receptors (CCR2 and CX3CR1), and colony-stimulating factor-1 (CSF1) mRNAs was carried out after laser capture microdissection of ED1+ and ED2+ cells.
LPS-treated pulps showed significant increases in (1) density of ED1+ and ED2+ cells beneath the amputation site and (2) expression levels of TLR4, CD14, CSF1, and CX3CR1 mRNAs, as compared with non-LPS-treated groups. CCR2 mRNA showed no significant difference between each group.
LPS treatment of cultured rat molars caused the accumulation of resident macrophages and enhanced the expression of TLR4, CD14, CSF1, and CX3CR1 mRNAs in these cells. Up-regulation of these molecules might be involved in the differentiation and subsequent migration of resident macrophages of the pulp.
Journal of endodontics 09/2011; 37(9):1258-63. · 2.95 Impact Factor
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ABSTRACT: This report describes non-surgical endodontic treatment of Oehlers' type III dens invaginatus in a maxillary lateral incisor with the aid of postobturation cone-beam computed tomography (CBCT). The endodontic treatment was initiated with the aid of a surgical operating microscope, and two canals, one of which represented the invagination, were instrumented, irrigated under passive ultrasonic activation and obturated with the lateral condensation technique. As postobturation periapical radiographs suggested the presence of untereated and/or unfilled areas in the root canal and invagination, CBCT was taken to assess the possibility of further treatment. The CBCT scans demonstrated inaccessible and unfilled canal and invagination areas because of complex internal morphology characterized by (i) C- or ring-shaped cross-sectional canal configuration with constrictions at different points in different root levels and (ii) a prominent intraradicular cavity that was communicated with the enamel-lined invagination and opened into the apical periodontium. Thus, it was judged that further endodontic treatment was not feasible. A 14-month follow-up revealed a satisfactory clinical and radiographic outcome, suggesting that the chemomechanical debridement may have sufficed to induce periapical healing. CBCT greatly helped the decision of avoiding further intervention that could have been difficult to negotiate.
Dental Traumatology 07/2011; 27(6):478-83. · 1.20 Impact Factor
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ABSTRACT: The aim of this study was to compare the effects of three brands of nickel-titanium (NiTi) rotary files with different designs on the initiation of apical root cracks when working short, at, and beyond the apical foramen.
One-hundred eight teeth with straight single canals were selected and mounted on resin blocks with simulated periodontal ligaments, and the apex was exposed. The teeth were divided into 9 groups of 12 teeth according to the NiTi rotary file type used (Profile [Dentsply Maillefer, Ballaigues, Switzerland], K3 [SybronEndo, West Collins, CA], and EndoWave [FKG Dentaire, La-Chaux-de-Fonds, Switzerland]) and working length (at CL, 1 mm short of [CL - 1 mm], and 1 mm beyond [CL + 1 mm] the apical foramen). Digital images of the apical surface of every tooth were taken during the apical enlargement sequence at each file change. These images were compared with the baseline image, and the presence of a crack was noted.
Significantly less cracks were observed in the CL - 1 mm group than in the CL and CL + 1 mm groups. No significant difference was found between the file types used.
Working 1 mm short of the apical foramen caused less cracks on the apical surface. In addition, more cracks were observed when using larger file sizes. Instrumentation with NiTi rotary files could potentially cause cracks on the apical root surface.
Journal of endodontics 04/2011; 37(4):522-5. · 2.95 Impact Factor
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ABSTRACT: We have identified tooth pulp-driven neurons (TPDNs) in the thalamic mediodorsal nucleus (MD) in rats and showed that the TPDNs' responsiveness in the MD is increased by chemical conditioning stimulation of allyl-isothiocyanate (mustard oil) to the molar tooth pulp. The aim of the present study was to address the role of N-methyl-d-aspartate receptors (NMDA receptors) in the sensitized central nervous system following the mustard oil application to the rat tooth pulp. Microinjection of MK-801, a noncompetitive NMDA receptor antagonist, to the thalamic MD nucleus reduced the TPDNs' responsiveness in the thalamic MD nucleus. Gene expression analysis showed that expression levels of NMDA receptor subunits NR2A and NR2D mRNAs in the thalamus were increased by the mustard oil application and that the increases were reduced by MK-801. When naloxone, an opioid receptor antagonist, was given systemically following the MK801 microinjection, the TPDNs' responsiveness was rekindled and expression levels of NR2D and NR2A mRNAs were increased. Moreover, lidocaine pretreatment abolished the mustard oil-induced upregulation of NR2D and NR2A mRNAs. These results suggest that, during central sensitization, interaction of NMDA receptors and endogeneous opioid-related inhibitory mechanisms plays critical role in the alteration of the TPDNs' responsiveness in the thalamic MD nucleus.
Brain research 01/2011; 1371:16-22. · 2.46 Impact Factor
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ABSTRACT: The immunostaining based Laser-capture microdissection (LCM) method called immune-LCM allows us to quantify the mRNA. Immune-LCM has recently been introduced to enhance identification of cells carrying a particular protein from frozen tissue samples. We have recently performed the immune-LCM of formaldehyde-fixated, paraffin-embedded tissues immunostained with a monoclonal antibody Factor VIII. This method could be useful for quantitative gene expression analysis in blood vessels from tumors of patients that have been treated with antiangiogenic drugs, allowing for validation of the effect of drug on the expected targets. Such capability might be exceedingly useful for the evaluation of the bioactivity of new drugs. This method is also useful to compare gene expression patterns in tumor cells versus endothelial cells during tumor progression or tumor angiogenesis.
Methods in molecular biology (Clifton, N.J.) 01/2011; 755:395-403.
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ABSTRACT: Pluripotent mesenchymal stem cells possess the ability to differentiate into many cell types, but the precise mechanisms of differentiation are still unclear. Here, we provide evidence that Rbpj (recombination signal-binding protein for immunoglobulin kappa j region) protein, the primary nuclear mediator of Notch, is involved in osteogenesis. Overexpression of Rbpj promoted osteogenic differentiation of mouse Kusa-A1 cells in vitro and in vivo. Transient transfection of an Rbpj expression vector into Kusa-A1 cells upregulated promoter activities of Runx2 and Ose2. Enhanced osteogenic potentials including high alkaline phosphatase activity, rapid calcium deposition, and increased calcified nodule formation, were observed in established stable Rbpj-overexpressing Kusa-A1 (Kusa-A1/Rbpj) cell line. In vivo mineralization by Kusa-A1/Rbpj was promoted compared to that by Kusa-A1 host cells. Histological findings revealed that expression of Rbpj was primarily observed in osteoblasts. These results suggest that Rbpj may play essential roles in osteoblast differentiation.
Biochemical and Biophysical Research Communications 09/2010; 400(1):39-45. · 2.48 Impact Factor
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ABSTRACT: The purpose of this study was to investigate the strain and temperature change in dentin induced by laser irradiation of human root canals with or without water cooling. Twenty-eight palatal roots of extracted human maxillary first molars were used. The strain in dentin was measured using strain gauges fixed on the apical third of the buccal root surface. The temperature change of the root dentin was monitored using thermocouples embedded into dentin near the apex. The root canal was irradiated with Er:YAG or Nd:YAG laser at an output of 1 W (100 mJ, 10 pps) for 5 s. The tip of the irradiation fibre was located 2.0 mm from the root apex. With water cooling, the mean maximum strain induced by Er:YAG laser was significantly lower than that by Nd:YAG laser (P < 0.05). However, without water cooling, no significant difference between the two lasers (P > 0.05) was found. The results suggest that the strain induced by Er:YAG laser irradiation in dentin with water cooling may be minimal, but there still might be a risk of root micro-fracture if cooling is not sufficient.
Australian Endodontic Journal 08/2010; 36(2):74-8. · 0.96 Impact Factor
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ABSTRACT: Notch signalling is of fundamental importance to various processes during embryonic development and in adults. The possible role of Hey1, an important Notch signalling component, in odontoblast differentiation was evaluated in this study. Primary cultured dental pulp cells, derived from upper incisors of 5-week-old Wistar rats, were placed in -modification of Eagle's minimal essential medium supplemented with 10% Fetal Bovine Serum (FBS), and ascorbic acid (AA) and β-glycerophosphate (β-GP), with or without dexamethasone, and cultured on dishes coated with collagen type IA for 7 days. Conventional and real-time Polymerase Chain Reaction (PCR) was performed to determine the expression of Notch-related genes and dentin sialophosphoprotein as a marker of odontoblast differentiation. Dentin sialophosphoprotein and Hey1 expression was significantly increased and decreased in the presence of AA + β-GP compared with controls, respectively. These findings suggest that Hey1 may be a negative regulator in odontoblast differentiation.
Australian Endodontic Journal 07/2010; 36(2):54 - 58. · 0.96 Impact Factor
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ABSTRACT: Matrix extracellular phosphoglycoprotein (MEPE) is an extracellular matrix protein that is mainly expressed in mineralizing tissues, including the dental pulp. The purposes of this study were to clarify the localization of MEPE in the tooth germ and to investigate the roles of MEPE in the differentiation of odontoblasts. The immunohistochemical staining in the tooth germ of the upper first molars of male Wistar rats (postnatal day 3) revealed that MEPE was mainly localized in odontoblasts during dentinogenesis. Stable MEPE-overexpressing and MEPE-knockdown cell lines, which were established in odontoblast-lineage cells (OLCs), showed lower and higher differentiation capabilities, respectively. Eukaryotic proteins of the N-terminal fragment of MEPE produced in HEK cells had no effect on the differentiation of OLCs, whereas the C-terminal fragment containing an RGD sequence inhibited their differentiation. These results indicated that the C-terminal fragment of MEPE containing an RGD sequence, cleaved in odontoblasts, appeared to be the active form of MEPE, which may play important roles in dentinogenesis and pulpal homeostasis by keeping the odontoblasts in immature condition.
Biochemical and Biophysical Research Communications 07/2010; 398(3):406-12. · 2.48 Impact Factor
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ABSTRACT: The aim of this study was to compare the fracture resistance of roots following root canal therapy using the RC Sealer system, the Epiphany system and the conventional system of gutta-percha and Sealapex. Fifty-six maxillary central incisors were divided into eight groups of seven teeth each, according to master apical file size and obturation systems. Obturation materials in the root canal were vertically loaded using a universal testing machine. Fracture loads were analysed by ANOVA and Tukey comparison, and fracture patterns were analysed with ordinal logistic regression. Master apical file size 80 had a significantly lower fracture load than size 40 (P < 0.05). The groups obturated using the Resilon Cone and the Epiphany Sealer had significantly lower fracture loads than the other groups (P < 0.05). There was no significant improvement in resistance to vertical root fractures using the examined adhesive resin root canal filling systems, compared with conventional gutta-percha and sealer.
Australian Endodontic Journal 04/2010; 36(1):19-23. · 0.96 Impact Factor
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ABSTRACT: The aim of this study was to investigate the safety of laser use under the dental microscope. Nd:YAG, Er:YAG and diode lasers were used. The end of the tips was positioned at a distance of 5 cm from the objective lens of a dental microscope. Each eye protector was made into a flat disc, which was fixed on the lens of the microscope. The filters were placed in front of the objective lens or behind the eye lens. Transmitted energy through the microscope with or without the filters was measured. No transmitted laser energy was detected when using matched eye protectors. Mismatched eye protectors were not effective for shutting out laser energy, especially for Nd:YAG and diode lasers. None or very little laser energy was detected through the microscope even without any laser filter. Matched filters shut out all laser energy irrespective of their positions.
Australian Endodontic Journal 04/2010; 36(1):6-11. · 0.96 Impact Factor
