Robert J. Loushine

Setting Properties and Cytotoxicity Evaluation of a Premixed Bioceramic Root Canal Sealer

INTRODUCTION This study investigated the setting time and micohardness of a premixed calcium phosphate silicate-based sealer (EndoSequence BC Sealer; Brasseler USA, Savannah, GA) in the presence of different moisture contents (0-9 wt%). The moisture content that produced the most optimal setting properties was used to prepare set EndoSequence BC Sealer for cytotoxicity comparison with an epoxy resin-based sealer (AH Plus; Dentsply Caulk, Milford, DE). METHODS Standardized disks were created with BC Sealer, AH Plus, Pulp Canal Sealer EWT (positive control) (SybronEndo, Orange CA), and Teflon (Small Parts Inc., Miami Lakes, FL; negative control). Disks were placed in Transwell Inserts, providing indirect contact with MC3T3-E1 cells. Succinate dehydrogenase activity of the cells was evaluated over a 6-week period using MTT ((3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Cytotoxicity profiles of BC Sealer and AH Plus were fitted with polynomial regression models. The time for 50% of the cells to survive (T(0.5)) was analyzed using the Wald statistic with a two-tailed significance level of 0.05. RESULTS BC Sealer required at least 168 hours to reach the final setting using the Gilmore needle method, and its microhardeness significantly declined when water was included in the sealer (P = .004). All set sealers exhibited severe cytotoxicity at 24 hours. The cytotoxicity of AH Plus gradually decreased and became noncytotoxic, whereas BC Sealer remained moderately cytotoxic over the 6-week period. A significant difference (P < .001) was detected between T(0.5) of BC Sealer (5.10 weeks; 95% confidence interval [CI], 4.69-5.42, standard error [SE] = 0.09) and T(0.5) of AH Plus (0.86 weeks; 95% CI, 0.68-1.05; SE = 0.18). CONCLUSIONS Further studies are required to evaluate the correlation between the length of setting time of BC Sealer and its degree of cytotoxicity.

Effects of different exposure times and concentrations of sodium hypochlorite/ethylenediaminetetraacetic acid on the structural integrity of mineralized dentin.

INTRODUCTION This study tested the null hypothesis that there is no difference between the use of 1.3% NaOCl/17% ethylenediaminetetraacetic acid (EDTA) and 5.25% NaOCl/17% EDTA irrigation regimens on the collagen degradation and flexural strength reduction in mineralized dentin. METHODS Dentin powder and mineralized dentin sections were immersed in 1.3% or 5.25% NaOCl for 10-240 minutes and then rinsed with 17% EDTA as the final irrigant for 2 minutes. Untreated mineralized dentin powder/sections served as controls in the respective experiments. Dentin powders were examined by using Fourier transform infrared (FT-IR) spectroscopy to analyze their relative subsurface intact collagen content with the apatite/collagen ratio. Hydrated dentin sections were subjected to 3-point flexure under water for determining their flexural strengths. RESULTS Collagen degradation was significantly increased and the flexural strength of mineralized dentin was significantly reduced after the use of 5.25% NaOCl as the initial irrigant for more than 1 hour (P < .05). Conversely, changes were insignificant when 1.3% NaOCl was used as the initial irrigant for up to 4 hours (Kruskal-Wallis analysis of variance, n = 10, P < .05). CONCLUSIONS The null hypothesis was rejected. The deleterious effects attributed to the use of NaOCl on dentin are concentration-dependent and time-dependent and are not associated with the demineralization caused by the use of EDTA as the final active irrigant.

Canal and isthmus debridement efficacies of two irrigant agitation techniques in a closed system

AIM To compare canal and isthmus debris debridement efficacies of the manual dynamic irrigation (MDI) and apical negative pressure (ANP) techniques in the mesial root of mandibular first molars with narrow isthmi, using a closed canal design. METHODOLOGY Micro-computed tomography was employed to select 20 teeth, each containing a narrow isthmus. Each root was sealed at the apex with hot glue and embedded in polyvinylsiloxane to simulate a closed canal system. The teeth were submitted to a standardized instrumentation protocol. Final irrigation was performed with either the MDI or the ANP technique using the EndoVac system (N=10). Masson trichrome-stained sections were prepared from completely demineralized roots at 10 canal levels between 1 and 2.8mm of the anatomical apices. Areas occupied by canals and isthmus of each root and debris in the corresponding regions were digitized by the NIH Image J software and statistically analysed using two-way repeated measures anova. RESULTS For the instrumented canals, there were no differences between the two groups (P=0.131) in the area occupied by debris at all canal levels (P=0.343). Conversely, for the isthmus, less debris was found in the ANP group (P<0.001) but no differences were seen in each group with respect to the 10 canal levels (P=0.352). CONCLUSION Neither technique completely removed debris from the isthmus regions. However, the EndoVac system, which encompasses the ANP concept, removed considerably more debris from narrow isthmi in mandibular mesial roots.

Sealing Ability of Mineral Trioxide Aggregate and Super-EBA When Used as Furcation Repair Materials: A Longitudinal Study

Immediate sealing of furcation perforations enhances the repair process. The purpose of this study was to longitudinally compare the ability of mineral trioxide aggregate (MTA) and Super-EBA to seal furcation perforations. Fifty-one extracted human maxillary molars were decoronated 3 mm above the CEJ, and the roots were amputated 3 mm below the furcation. A #2 high-speed round bur was used to perforate the center of the furcations. The canals were obturated with gutta-percha, and the root ends were sealed with C&B Metabond. Three experimental groups of 15 teeth each were restored with MTA, Super-EBA, or a combination of MTA in the perforation and a Super-EBA dome on the pulpal floor. Six teeth served as controls. Each tooth was affixed to a fluid filtration device and subjected to a pressure of 20 cm H2O. The integrity of the perforation seal was evaluated initially at 30 min for the Super-EBA and the combination groups and at 4 h for the MTA group. Additional measurements were then made at 24 h, 1 week, and 1 month. The controls behaved as expected. A two-way ANOVA revealed a significant difference (p = 0.01) between materials. Tukeys test isolated the difference to Super EBA as producing a superior seal but only at 24 h. There was no significant effect with time (p = 0.57) or the interaction of the materials with time (p = 0.66). All materials sealed the perforations very well. The maximum leakage of all materials was <0.007 microL min(-1) cm H2O(-1).

Effects of eugenol on resin bond strengths to root canal dentin.

The purpose of this study was to compare the regional bond strengths of C&B Metabond resin to root canal dentin, with or without treatment using a eugenol-containing endodontic sealer liquid. Eighteen extracted human canines were decoronated at the cementoenamel junction with a slow speed saw. The apical third of the root was removed leaving the cervical and middle dentin. The canal space was then enlarged with files, Gates-Glidden burs, and parapost drills. The teeth were ground on either the mesial or distal sides, permitting direct access to the entire canal. The cervical or middle third dentin was treated with Kerr Root Canal Sealer liquid, alternating between the middle and cervical thirds. Each tooth served as its own control. The adhesive resin was then luted directly to the prepared canal. Specimens, 1 x 1 x 8 mm, were prepared and mounted to a Vitrodyne testing machine enabling microtensile bond strengths to be measured. Data were analyzed using a two-way ANOVA and the least squares means test. The mean microtensile bond strengths for the cervical and middle third dentin treated with eugenol were 13.6 +/- 6.1 MPa (n = 33) and 14.8 +/- 3.9 MPa (n = 29), respectively. Without the eugenol, the mean bond strengths were 18.1 +/- 6.0 MPa (n = 31) and 17.3 +/- 4.6 MPa (n = 31) for the cervical and middle sections. The specimens treated with the eugenol liquid had significantly lower bond strengths than those without eugenol (p < 0.05) only in the cervical third. The region of the tooth tested had no effect on bond strength. That is, bond strength of the cervical third was not significantly different from bond strength on the middle third in either of the two groups (with or without eugenol) tested.

Interpretation of periapical lesions comparing conventional, direct digital, and telephonically transmitted radiographic images

The purpose of this study was to evaluate direct digital radiography (DDR) and telephonically transmitted images versus conventional radiography in the interpretation of artificial periapical bone lesions. Five teeth were chosen from four cadaver jaw specimens. Three types of DDR images were evaluated: DDR stored images, DDR transmitted images, and DDR reversed images. A total of 150 DDR computer monitor images and 56 D-speed film images were evaluated by three endodontists and one endodontic graduate student. The Wilcoxon signed-ranks test was used for statistical analyses of the results. DDR reversed images were statistically inferior to DDR stored images, DDR transmitted images, and conventional radiography images (p < 0.0001). There were no statistically significant differences between DDR stored images, DDR transmitted images, and conventional film images in the ability of the evaluator to identify artificial periapical bone lesions (p > 0.05).

In vitro bacterial penetration of endodontically treated teeth coronally sealed with a dentin bonding agent.

This study evaluated the effectiveness of a dentin bonding agent as a barrier to prevent coronal microleakage and examined the effect of a eugenol-based sealer on the sealing ability of this resin adhesive. Fifty-one extracted human mandibular molars were incorporated in a model system using an oral streptococci as a microbial marker. Group 1 consisted of 15 teeth that were obturated with only gutta-percha and received a coronal barrier of Clearfil Liner Bond 2V. Group 2 was identical to group 1, but included the use of a eugenol-based sealer in the obturation. Group 3 consisted of 15 teeth that were obturated with gutta-percha and sealer, but did not receive a coronal barrier. Six teeth served as controls. Bacterial penetration was monitored for 90 days. Results were analyzed after 30, 60, and 90 days with Fishers exact test (p < 0.05). All controls behaved as expected. Neither group 1 nor group 2 exhibited any bacterial leakage. Eleven of the 15 specimens in group 3 leaked between 15 and 76 days. The coronal barriers in group 1 and group 2 were significantly better in preventing coronal microleakage at 60 days (p = 0.002) and 90 days (p = 0.00005). The presence of eugenol in the sealer had no significant effect on the sealing ability of Clearfil Liner Bond 2V (p = 1).

Effect of varying the depth of heat application on the adaptability of gutta-percha during warm vertical compaction.

The purpose of this study was to compare the adaptability of gutta-percha after varying the depth of heat application in the obturation of a set of standard root canals. A split-tooth model was constructed using a human maxillary central incisor. The root canal was cleaned and shaped using a step-back preparation to a size #60 FlexOFile at the working length (WL). Five shallow depressions were produced on the root canal wall. Twenty obturations without sealer were performed for each technique (thermoplasticized injectable (TI), lateral condensation, and warm vertical compaction (WVC) with heat applications at 3, 4, 5, and 7 mm from the WL). After each obturation the model was separated and the mesial and distal sides of each obturation were examined and videotaped at x 32 magnification. The quality of the obturation was graded based on the replication to the WL, replication of the artificial depressions, surface adaptation, and homogenicity of the gutta-percha. The Kruskal-Wallis analysis and the Student-Newman-Keuls tests indicated that all the techniques were significantly different from each other (p < 0.05) except for the TI group versus the WVC group with the heat application to within 3 mm from the WL (p > 0.05). The TI technique was ranked best followed by the WVC with heat applications at 3, 4, 5, and 7 mm. The lateral condensation technique received the lowest ranking.

Resistance of a 4-META-containing, methacrylate-based sealer to dislocation in root canals.

The dislocation resistance of root fillings created with MetaSEAL, a self-adhesive 4-META-containing methacrylate resin-based sealer, was evaluated. Forty-six incisors were cleaned and shaped using NaOCl and EDTA as irrigants. They were filled with gutta-percha/MetaSEAL or gutta-percha/AH Plus sealer using either a single-cone technique or warm vertical compaction (n = 10). The roots were sectioned at the coronal and middle thirds to obtain thin slices, which were subjected to compressive loading to displace the set sealer/filling toward the coronal side of the slice. The remaining six teeth were filled with gutta-percha/MetaSEAL and cryofractured for scanning electron microscopic examination. The push-out strength of AH Plus was significantly higher than MetaSEAL irrespective of filling techniques (p < 0.05). A minimal hybrid layer was seen in radicular dentin, and resin tags were inconsistently identified from canal walls in the MetaSEAL-filled canals. The lower dislocation resistance in MetaSEAL-filled canals challenges the use of a self-adhesive bonding mechanism to create continuous bonds inside root canals.

Effects of Calcium Silicate–based Materials on Collagen Matrix Integrity of Mineralized Dentin

INTRODUCTION Calcium silicate-based materials (CSMs) are used in various endodontic procedures. The present study examined whether prolonged contact of mineralized dentin with recently commercialized versions of these materials adversely affects dentin collagen matrix integrity. METHODS Dentin slabs prepared from extracted human third molars (7 × 3 × 0.3 mm) were divided into 3 groups on the basis of the material to which dentin was exposed (MTA Plus, Biodentine, untreated control dentin slabs) and the time period of exposure (24 hours, 1, 2, and 3 months; n = 6). Hydroxyproline assay was performed on each groups supernatant to quantify the collagen extraction amounts of each group per time period. Data were analyzed with two-factor repeated-measures analysis of variance and Holm-Sidak pair-wise comparisons (α = 0.05) to determine the effects of material and aging time on collagen extraction. Dentin slabs from the 3 months of aging group were demineralized for transmission electron microscopy examination of collagen matrix ultrastructural changes. RESULTS Material (P = .002), aging time (P < .001), and their interactions (P = .007) significantly affected the amount of hydroxyproline (pg/mg of mineralized dentin) extracted from mineralized dentin and were significantly correlated by power regression models. Collagen degradation was identified from the surface of dentin slabs that were in direct contact with CSMs. CONCLUSIONS Prolonged contact of mineralized dentin with CSMs has an adverse effect on the integrity of the dentin collagen matrix. However, the amount of collagen extracted was limited to the contact surface. Clinicians can continue to apply CSMs in endodontic procedures; however, caution is advised when these materials are applied to thin dentinal walls.