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.

Bonding of self-adhesive (self-etching) root canal sealers to radicular dentin.

The latest generation of methacrylate resin-based sealers has eliminated the use of separate self-etching primers by incorporating acidic resin monomers in the sealers to render them self-adhesive to dentin. This study examined the adhesive strengths, interfacial ultrastructure, and tracer penetration of a nonetching (EndoREZ; Ultradent, South Jordan, UT) and two self-adhesive methacrylate resin-based sealers (MetaSEAL; Parkell, Farmington, NY, and RealSeal SE; SybronEndo, Orange, CA) when they were applied to radicular dentin following the manufacturers' recommended use of EDTA as the active final rinse. A modified push-out testing design was used to evaluate the dislodgement of core-free sealers. The mixed sealers were placed in dimensionally identical, artificially created canal spaces prepared in the coronal, middle, and apical thirds of radicular dentin. After setting, each sealer-filled cavity was subjected to compressive loading until failure. Additional specimens were prepared for transmission electron microscopy to examine the ultrastructure and nanoleakage within the sealer-radicular dentin interface. The two self-adhesive sealers MetaSEAL and RealSeal SE exhibited higher push-out strengths than the nonetching sealer EndoREZ when EDTA was used as the active final rinse. All three sealers showed a 1- to 1.5-microm thick zone of partially demineralized dentin, with the EDTA dentin demineralization effect masking the true self-etching potential of MetaSEAL and RealSeal SE. The true self-etching potential of self-adhesive sealers is a clinically important attribute that should be further investigated. Incomplete smear layer removal from the apical third of instrumented canal walls may jeopardize the performance of self-adhesive sealers should they fail to self-etch without the adjunctive use of calcium chelating irrigants.

Contemporary methacrylate resin-based root canal sealers exhibit different degrees of ex vivo cytotoxicity when cured in their self-cured mode.

The cytotoxicity of four methacrylate resin-based sealers was investigated by the 3-(4,5-dimethyl-thiazoyl)-2,5-diphenyl-tetrazolium bromide assay, which measures cell viability by assessing its succinate dehydrogenase activity. The sealers were polymerized in the self-cured mode to simulate the setting condition upon their extrusion into periradicular tissues. Disks were prepared from EndoREZ (Ultradent, South Jordan, UT), RealSeal (SybronEndo, Orange, CA), MetaSEAL (Parkell, Farmington, NY), and RealSeal SE (SybronEndo) together with positive and negative controls. After setting, they were placed in direct contact with rat osteosarcoma (ROS 17/2.8) cells and for 5 succeeding weeks after immersing in simulated body fluid (SBF). All sealers exhibited severe toxicity initially (week 0). EndoREZ and RealSeal remained severely toxic after five cycles of SBF immersion. Toxicity of the two self-etching resin-based sealers MetaSEAL and RealSeal SE decreased gradually over time. Transmission electron microscopy of cells exposed to RealSeal SE showed variable degrees of cell injury that reflect its toxicity status. Cells with intact mitochondria were identifiable after the sealer became noncytotoxic at week 5.

Hybrid Root SEAL (MetaSEAL) creates hybrid layers in radicular dentin only when EDTA is used as the final rinse.

PURPOSE: To test if the hybrid layer formation by Hybrid Root SEAL (Sun Medical Co.), a 4-META-containing auto-adhesive self-etching root canal sealer, is affected by the sequence of irrigants employed for removing canal wall smear layers during root canal treatment. METHODS: Single-rooted teeth were shaped and irrigated with EDTA as initial rinse/NaOCl as active final rinse (Group 1), or NaOCl as initial rinse/EDTA as active final rinse (Group 2). All canals were obturated with Hybrid Root SEAL using a single-cone technique. Root slices derived from the coronal, middle and apical thirds of the roots were processed for transmission electron microscopy after removing the gutta-percha, leaving the sealer intact. Additional filled canals from the two groups were evaluated for fluid leakage. RESULTS: Hybrid layer was absent in Group 1 and was present only when a collagen matrix was produced by EDTA demineralization (Group 2). Significantly more leakage (4.03 +/- 1.94 microL min(-1) vs. 1.50 +/- 0.42 microL min(-1); P < 0.05) was observed in the absence of dentin hybridization.

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.

Calcium phosphate phase transformation produced by the interaction of the portland cement component of white mineral trioxide aggregate with a phosphate-containing fluid.

The bioactivity of mineral trioxide aggregate (MTA) has been attributed to its ability to produce hydroxyapatite in the presence of phosphate-containing fluids. It is known that stoichiometric hydroxyapatites do not exist in biological systems and do not contribute to the osteogenic potential of calcium phosphate-based biomaterials. Because Portland cement is the active ingredient in white MTA, we have characterized the calcium phosphate phases produced when set white Portland cement was immersed in phosphate-buffered saline using pH and turbidity measurements, scanning electron microscopy, energy dispersive X-ray analysis, transmission electron microscopy, electron diffraction, x-ray diffraction, and Fourier transform-infrared spectroscopy. An amorphous calcium phosphate phase was initially formed that transformed to an apatite phase, with the latter consisting of calcium-deficient, poorly crystalline, B-type carbonated apatite crystallites. Amorphous calcium phosphate is a key intermediate that precedes biological apatite formation in skeletal calcification. Thus, the clinical manifestations of bioactivity with the use of MTA may at least be partially attributed to the mineralization induction capacity of its Portland cement component.

Evaluation of the quality of the apical seal in Resilon/Epiphany and Gutta-Percha/AH Plus-filled root canals by using a fluid filtration approach.

This in vitro fluid filtration study compared the apical leakage of root canals that were obturated with Resilon/Epiphany (Resilon Research LLC, Madison, CT) or gutta-percha/AH Plus (GP/AH; DENTSPLY Tulsa, Tulsa, OK) sealer using warm vertical condensation. Fluid flow rate through the filled roots was measured 7 days after the obturations by using a fluid filtration device. Measurements were made before root resection and after 3 to 11 mm of resections. Before any removal of root length, there were no significant differences between the roots filled with Resilon/Epiphany or GP/AH. Analysis of individual root resection results revealed that differences in the fluid flow rate of the two systems occurred only when 9 or 10 mm of the roots were resected with more than half the root length resected when GP/AH Plus leaked more than Resilon/Epiphany. There were no significant differences in the fluid flow rate up to and including 8 mm of root resection. It is concluded Resilon/Epiphany sealed 17-mm root canals as well as gutta-percha and AH Plus sealer and that it does not create a monoblock root filling that does not leak.

Susceptibility of a polycaprolactone-based root canal-filling material to degradation. III. Turbidimetric evaluation of enzymatic hydrolysis.

The susceptibility of Resilon (Pentron Clinical Technologies, Wallingford, CT) to biotic biodegradation by two hydrolases, lipase PS and cholesterol esterase, was investigated with a turbidimetric approach by measuring the optical density reductions in aqueous emulsions containing dissolved, filtered, surfactant-stabilized polymeric components of Resilon. Polycaprolactone, the major polymeric component of Resilon, was also examined in a similar manner using equivalent or a four-fold increase in enzyme concentration. Optical density time plots were characterized by an initially linear steep reduction in optical density, from which the reaction rates were derived. For both enzymes, the rates of hydrolysis for Resilon were much faster than those of polycaprolactone at 1x or even 4x enzyme concentration. Field-emission scanning electron microscopy of air-dried Resilon and polycaprolactone emulsions revealed the presence of spherical polymer droplets that appeared deformed, pitted, or much reduced in dimensions after enzymatic hydrolysis.

Metrologic evaluation of the surface of white MTA after the use of two endodontic irrigants.

This study examined the effects of calcium-depleting endodontic irrigants, ethylenediamine tetra-acetic acid (EDTA) and BioPure MTAD, on the dissolution, surface characteristics, and ultrastructural characteristics of white mineral trioxide aggregate (MTA). The latter was mixed in a 0.35 water-cement ratio, condensed into cylindrical wells created in Plexiglas platforms, and allowed to harden completely before initial treatment with 1.3% NaOCl and final treatments with either 17% EDTA for 5 minutes, or BioPure MTAD for 1, 3, or 5 minutes. Analysis of the mean depths of material removed using three-dimensional profilometry revealed significantly more material removed by BioPure MTAD. Although these minor depth changes (<10 microm) are unlikely to cause clinical concern, the BioPure MTAD-treated MTA surfaces exhibited higher surface roughness and with more calcium extracted when compared with EDTA treatment. Decomposition of particle-binding hydration phases by acid corrosion raises potential concern on the strength and sealing properties of MTA-repaired perforations following final irrigation by BioPure MTAD.

Self-etching adhesives increase collagenolytic activity in radicular dentin.

Endogenous matrix metalloproteinases (MMPs) release from crown dentin and their activation results in degradation of hybrid layers created by dentin adhesives. This study tested the hypothesis that instrumented intraradicular dentin possesses latent collagenolytic activity that is activated by mild self-etching adhesives. Root dentin shavings were produced from 50 cleaned and shaped, saline-irrigated root canals using Gates Glidden drills and rinsed with sodium azide to prevent bacterial growth. Dried dentin powder aliquots were treated with two clinically-relevant MMP inhibitors, 2% chlorhexidine for 10 minutes and 17% EDTA for 1 minute. Additional dentin powder was mixed with Clearfil Liner Bond 2V or Clearfil Tri-S Bond for 1 minute followed by extracting the adhesives with acetone. Dentin powder was also treated with 2% chlorhexidine for 10 minutes before or after adhesive application. Collagenolytic activities of the nine groups were assayed with a fluorometer in 96-well plates, by recording the changes in fluorescence before and after addition of fluorescein-labeled type I collagen. Epoxy resin-embedded powders were examined with TEM for the extent of demineralization. Instrumented, mineralized intraradicular dentin possessed low but detectable collagenolytic activity that was inhibited by chlorhexidine (p < 0.001) and EDTA (p < 0.001). Both adhesives partially demineralized the dentin powder and activated latent MMPs, with 14- to 15-fold increases in collagenolytic activities (p < 0.001) that were significantly (p < 0.001) but incompletely inactivated after 10 min application of chlorhexidine. Mild self-etching adhesives activate latent MMPs without denaturing these enzymes, and may adversely affect the longevity of bonded root canal fillings and posts.

Improving the performance of EndoRez root canal sealer with a dual-cured two-step self-etch adhesive. I. Adhesive strength to dentin.

This study examined the hypotheses that the use of a self-etching-adhesive improves the adhesion of EndoRez to dentin, and that both techniques are unaffected by the prolonged use of NaOCl when EDTA is used as the final rinse. A tensile bond testing design was employed with composite cylinders bonded to dentin using: (a) 17% EDTA-2 min/EndoRez; (b) 6.15% NaOCl-1 h/EDTA-2min/EndoRez; (c) EDTA-2 min/Clearfil Liner Bond 2V/EndoRez; and (d) NaOCl-1 h/EDTA-2min/Clearfil Liner Bond 2V/EndoRez. Fractographic analyses of debonded specimens were performed using SEM. Two-way ANOVA and Tukey test revealed higher tensile bond strengths with the self-etching-adhesive-modified EndoRez technique (p < 0.001). Sealer tags pulled away from dentinal tubules when EndoRez was used alone, whereas mixed failures occurred within sealer tags, hybrid layers, adhesive, and sealer in the self-etching-adhesive-modified EndoRez technique. Both techniques were unaffected by the prolonged use of NaOCl (p > 0.05) when EDTA was employed as the final rinse.

Improving the performance of EndoREZ root canal sealer with a dual-cured two-step self-etch adhesive. II. Apical and coronal seal.

This study tested the hypothesis that the seal of EndoREZ may be improved with an adhesive-modified technique. Instrumented single-rooted teeth were filled with: (a) Warm vertical compaction with AH Plus (control); (b). EndoREZ with master cones and passive application of accessory cones; (c) Application of Clearfil Liner Bond 2V before EndoREZ and gutta-percha condensation. Leakage was assessed by fluid filtration at 10 psi before root resection and after 3 to 12 mm apical resections. Statistical analyses revealed significant differences in filling technique and resection lengths. EndoREZ exhibited significantly higher overall leakage, while no difference was found between AH Plus and the adhesive-modified EndoRez technique. Apical resection of EndoREZ to 12 mm exhibited more leakage than all other interactions, but was not significantly different from the same material resected to 9 mm. Although EndoREZ exhibited an acceptable apical seal, its coronal seal may be improved with the use of a dual-cured self-etch adhesive.

A comparison of cohesive strength and stiffness of Resilon and gutta-percha.

The purpose of this study was to compare the cohesive strength and stiffness of Resilon and gutta-percha under dry conditions and after 1 month of water storage to determine if they are stiff enough to reinforce roots. Dog-bone shaped specimens were created and pulled to failure at 3 mm/min in a Vitrodyne universal tester. The apparent modulus of elasticity, percent elongation, and yield stress were measured. Cohesive strength was defined as the yield stress. The data were analyzed using a two-way ANOVA for each outcome (alpha = 0.05). The results of this study show that the cohesive strength (that is the tensile stress when they begin to flow or break) and modulus of elasticity (or stiffness) of gutta-percha and Resilon are relatively low. Although the results showed statistically significant differences between the physical properties of gutta-percha and Resilon under varied conditions, these differences were not clinically significant. In conclusion, the stiffness of Resilon and gutta-percha is too low to reinforce roots after root canal therapy.

Is an oxygen inhibited layer required for bonding of resin-coated gutta-percha to a methacrylate-based root canal sealer?

Removal of the oxygen inhibition layer from the surface of resin-coated gutta-percha cones during packaging has been hypothesized for their weak adhesion to a methacrylate-based root canal sealer, resulting in their frequent delamination from the sealer after root canal obturation. This study examined the feasibility of creating oxygen inhibition layers on resin-coated gutta-percha cones via the adjunctive application of a dual-cured dentin adhesive just before bonding. Composite cylinders were bonded with EndoREZ to flat, resin-coated gutta-percha disks and similar disks that were post-treated with Prime&Bond NT Dual Cure adhesive and stressed to failure using a modified microshear testing design. Although shear strengths for both groups were relatively low, a 5-fold increase in shear strength was observed after adhesive application, with complex interfacial failures instead of complete sealer delamination from the resin-coating. In-situ dentin adhesive application appears to have merits in enhancing the coupling of resin-coated gutta-percha to methacrylate sealers.

Ultrastructure of intraradicular dentin after irrigation with BioPure MTAD. II. The consequence of obturation with an epoxy resin-based sealer.

In addition to an initial report on the creation of thick demineralized collagen matrices in root dentin that were irrigated with BioPure MTAD or EDTA as final rinses for 5 minutes, this study examined the effect of a 2-minute irrigation time of these final irrigants on the extent of demineralization of intact intraradicular dentin and on interfacial nanoleakage after obturation with AH Plus and gutta-percha using warm vertical compaction. Transmission electron microscopy confirmed that the reduction in irrigation time did not compromise the smear layer removing capacity of these two irrigants when 1.3% NaOCl was employed as the initial rinse. BioPure MTAD created 5 to 6 microm thick demineralized dentin matrices, while 17% EDTA produced 1 to 2 microm thick matrices that were suboptimally infiltrated by the epoxy resin-based sealer, as manifested by the occurrence of silver tracer deposits within the hybrid layers.

Ultrastructure of smear layer-covered intraradicular dentin after irrigation with BioPure MTAD.

The structure of mechanically instrumented intraradicular dentin after irrigation with NaOCl as the initial rinse and BioPure MTAD as the final rinse were examined from the coronal, middle, and apical parts of root canal walls using transmission electron microscopy. Sterile distilled water and EDTA as final rinses were employed as the respective positive and negative controls under the same experimental conditions. There were 2 to 5 mum thick smear layers produced on mechanically instrumented root canal walls that were completely removed by EDTA and BioPure MTAD under agitation. Both irrigants created a zone of demineralized collagen matrices in eroded dentin and around the dentinal tubules, with the mildly acidic BioPure MTAD being more aggressive than EDTA. These demineralized dentin zones create the opportunity for dentin hybridization by infiltration of hydrophilic adhesives/sealers. However, the potential consequences of compaction of hydrophobic sealers against air-dried, collapsed collagen matrices, and hydrolytic degradation of incompletely infiltrated matrices remain unresolved.

Bondability of Resilon to a methacrylate-based root canal sealer.

Resilon broadens the dimensions of endodontic adhesion by introducing the possibility of creating monoblocs between root canal filling materials and intraradicular dentin by using methacrylate-based sealers. The adhesive strength of Resilon to RealSeal, a methacrylate-based root canal sealer, was evaluated using a modified microshear bond testing design. Flat Resilon surfaces with different roughness were created for bonding to the sealer and compared to a composite control. The composite control exhibited mean shear strength 7.3 to 26.9 times higher than those of the Resilon groups. Shear strength differences among the Resilon groups of different surface roughness highlighted the contribution of micromechanical versus chemical coupling in sealer retention. Ultrastructural evidence of phase separation of polymeric components in Resilon suggested that the amount of dimethacrylate incorporated into this filled, thermoplastic composite may not yet be optimized for effective chemical coupling to methacrylate-based sealers.

Application of biologically-oriented dentin bonding principles to the use of endodontic irrigants.

PURPOSE: To compare the removal of smear layer and the structure of the hybrid layer formed after the use of EDTA or MTAD solutions when used as a final flush. METHODS: Single-rooted extracted premolars (n=18) were collected and treated for root canal therapy using NaOCl irrigation followed by a final rinse of 17% EDTA, Biopure MTAD, or saline (negative control). The roots were obturated with gutta-percha and a hydrophilic HEMA-containing root canal sealer. The TEM specimens were impregnated with 50% silver nitrate to visualize sealing imperfections and nanoleakage. The structure of the coronal, middle and apical parts of root canal walls was examined using transmission electron microscopy. RESULTS: After NaOCl irrigation, a final rinse with BioPure MTAD or 17% EDTA completely removed the 2 microm-thick smear layer on mechanically instrumented root canal walls. The BioPure MTAD hybrid layer was thicker than the 17% EDTA hybrid layer. Both the BioPure MTAD and EDTA caused a collapse of the dentin matrix structure which impeded sealer infiltration and the formation of high quality hybrid layer bonding. The hybrid layers created in smear layer-covered dentin exhibited less potential for nanoleakage than the MTAD or EDTA hybrid layers.

Susceptibility of a polycaprolactone-based root canal filling material to degradation. II. Gravimetric evaluation of enzymatic hydrolysis.

Polycaprolactone is susceptible to enzymatic biodegradation via ester bond cleavage. This study examined the susceptibility of Resilon, a polycaprolactone-based root filling material to enzymatic hydrolysis. Resilon, gutta-percha, and polycaprolactone disks, prepared by compression molding, were incubated in phosphate-buffered saline, lipase PS or cholesterol esterase at 37 degrees C for 96 h. They were retrieved at different time intervals for gravimetric analysis and scanning electron microscopy. The materials exhibited slight weight gains when incubated in phosphate-buffered saline that can be attributed to water sorption. Gutta-percha showed similar weight gains in the two enzymes. Conversely, Resilon and polycaprolactone exhibited extensive surface thinning and weight losses after incubation in lipase PS and cholesterol esterase. Glass filler particles in Resilon were exposed following surface dissolution of the polymer matrix, creating rough surface topography. Biodegradation of Resilon by bacterial and salivary enzymes warrants further investigation of their activities using cultures of endodontically relevant microbes and human saliva extracts.

Effectiveness of resin-coated gutta-percha cones and a dual-cured, hydrophilic methacrylate resin-based sealer in obturating root canals.

The introduction of a polybutadiene-diisocyanate-methacylate resin-coating of gutta-percha enables the polyisoprene to be chemically coupled to methacrylate-based resin root canal sealers. This study examined the effectiveness of using passively fitting cones of this type of gutta-percha with a dual-cured version of EndoREZ sealer in obturating cleaned and shaped root canals. The hydrophilic nature of the sealer enabled the creation of an extensive network of 800 to 1200 mum long sealer resin tags after removal of the endodontic smear layer. Although no adhesive was employed, thin hybrid layers in root dentin were observed when EDTA was used as the final rinse. Nevertheless, interfacial gaps and silver leakage could be observed along the sealer-dentin interfaces that might be attributed predominantly to polymerization shrinkage of the sealer. Gaps and silver leakage were also identified between the gutta-percha resin-coating and the sealer.