Acid and Microhardness of Mineral Trioxide Aggregate and Mineral Trioxide Aggregate–like Materials

Introduction

The aim of this study was to compare the surface microhardness of BioAggregate, ProRoot MTA, and CEM Cement when exposed to an acidic environment or phosphate-buffered saline (PBS) as a synthetic tissue fluid.

Methods

Ninety cylindrical molds made of polymethyl methacrylate with an internal diameter of 6 mm and height of 4 mm (according to ASTM E384 standard for microhardness tests) were fabricated and filled with BioAggregate (n = 30), tooth-colored ProRoot MTA (n = 30), or CEM Cement (n = 30). Each group was then divided into 3 subgroups of 10 specimens consisting of those exposed to distilled water, exposed to PBS (pH = 7.4), or exposed to butyric acid (pH = 5.4). After 1 week the Vickers surface microhardness test was performed. Statistical analysis included 2-way analysis of variance, followed by post hoc Dunnett T3 in cases with lack of homoscedasticity and Tukey honestly significant difference in cases with homoscedasticity.

Results

The indentations obtained from the CEM Cement specimens exposed to an acidic pH were not readable because of incomplete setting. There was a significant difference between the microhardness of the materials regardless of the environmental conditions (P < .001). In all the environmental conditions, MTA had significantly higher and CEM Cement had significantly lower microhardness values (P < .001). All experimental cements had significantly higher microhardness values when exposed to PBS (P < .001) and had significantly lower microhardness values when exposed to butyric acid (P < .001).

Conclusions

The surface microhardness of BioAggregate, ProRoot MTA, and CEM Cement was reduced significantly by exposure to butyric acid and increased significantly by exposure to PBS. In all environmental conditions, MTA had significantly higher microhardness values.     

Properties of an Accelerated Mineral Trioxide Aggregate–like Root-end Filling Material

The purpose of the study was to examine the physicochemical properties of mineral trioxide aggregate (MTA)-like cements prepared from SiO₂, CaO, and Al₂O₃ as matrices, and ZnO, MgO, and Fe₂O₃ as additives. The MTA-like cements showed a significantly (p < 0.05) faster setting time (<14 minutes) than MTA when mixed with water, although there was a significant decrease (p < 0.05) in diametral tensile strength, ranging from 0.9 to 3.1 MPa in comparison to MTA (4.4 MPa). The addition of ZnO, MgO, and Fe₂O₃ to the MTA-like cement led to a significantly (p < 0.05) higher strength than MTA-like cement without additives. The pH values of the MTA-like cements were changed from an initial 11 to a high of 13, similar to the results of MTA. Future characterization of the examined MTA-like cement with shortened setting time is needed before it is considered a viable candidate for dental root-end–filling applications.     

Retrievability of Calcium Silicate–based Root Canal Sealers During Retreatment: An Ex Vivo Study

IntroductionThe retrievability of calcium silicate–based sealers (CSSs) during nonsurgical retreatment has been equivocal. This study compared the retrievability of 3 different CSSs using 1 of 3 different solutions or no solution.MethodsA total of 130 extracted teeth with a straight canal were decoronated to a standardized root length. The canals were instrumented to 35/.04 and divided into 3 groups (BC: EndoSequence BC sealer; EBC: EdgeBioceramic; NEO: NeoSEALERFlo), and each group was further divided into 4 subgroups (6% sodium hypochlorite; 5% acetic acid; carbonated water; no solution). After sealer placement, each master gutta-percha cone was placed intentionally 2 mm short of the working length (WL) to ensure the apical 2 mm was filled only with sealer. After storage for 21 days at 37°C and 100% humidity, retreatment was performed until apical patency was obtained. The operator was blinded to the CSS and solution used. Data were analyzed using analysis of variance to compare apical patency rates and the mock chair-times.ResultsOverall success rates for apical patency in BC, EBC, and NEO were 63.64%, 69.77%, and 100%, respectively. There was significantly higher apical patency rate in NEO than BC and EBC. The chair-time for NEO was significantly shorter than BC (P < .05) and EBC sealer (P < .001). There was no significant difference in the chair-time between BC and EBC sealer groups.ConclusionBC, EBC, and NEO sealers in a straight canal were consistently retrievable when no solution was used. Compared with no solution, the retrievability of BC, EBC, and NEO decreased when solutions were used.     

In Vitro Fracture Resistance of Roots Obturated with Epoxy Resin–based,Mineral Trioxide Aggregate–based,and Bioceramic Root Canal Sealers

Introduction

The aim of this study was to evaluate the fracture resistance of teeth filled with 3 different endodontic sealers.

Methods

Seventy-five single-rooted extracted mandibular premolars were decoronated to a length of 13 mm. The teeth were randomly divided into 5 groups (n = 15 for each group). In group 1, the teeth were left unprepared and unfilled (negative control), and in group 2, the teeth were left unobturated (positive control). The rest of the roots were prepared by using the ProTaper System up to a master apical file size of F3: group 3, bioceramic sealer (Endosequence BC sealer) + gutta-percha; group 4, mineral trioxide aggregate–based sealer (Tech Biosealer Endo) + gutta-percha; and group 5, epoxy resin–based sealer (AH Plus Jet) + gutta-percha. All root specimens were stored for 2 weeks at 100% humidity to allow the complete setting of the sealers. Each specimen was then subjected to fracture testing by using a universal testing machine at a crosshead speed of 1.0 mm/min⁻¹ until the root fractured. The force required to fracture each specimen was recorded, and the data were analyzed statistically.

Results

The fracture values of groups 3 and 5 were significantly higher than those of group 4 (P < .05). There was no significant difference between groups 3 and 5 (P > .05).

Conclusions

In contrast to Tech Biosealer Endo, Endosequence BC and AH Plus Jet sealer increased the force to fracture in root-filled single-rooted premolar teeth.     

Mercury Intrusion Porosimetry and Assessment of Cement-dentin Interface of Anti–washout-type Mineral Trioxide Aggregate

Introduction

One of the disadvantages of mineral trioxide aggregate (MTA) is washout (ie, the tendency of freshly prepared cement paste to disintegrate upon early contact with physiological fluids). A novel MTA (MTA Plus; Prevest Denpro, Jammu City, India) exhibits low washout and superior physical properties when mixed with a gel instead of water. When used as a root-end filler, MTA is in contact with both bone and root dentin. This study aimed to investigate the porosity and interfacial characteristics of the novel MTA mixed with water or antiwashout gel.

Methods

Porosity was evaluated after 1 or 28 days of immersion in Hank's balanced salt solution using mercury intrusion porosimetry. The root dentin to material interface was investigated using a scanning electron microscope and energy-dispersive X-ray spectroscopy complete with line scans and elemental maps.

Results

Anti–washout-type MTA Plus was found to have lower initial porosity than MTA Plus mixed with water although this trend was reversed after 28 days of immersion in physiological fluid. Both materials exhibited good marginal adaptation. The diffusion of silicon, calcium, and phosphorus across the cement/dentin interface was observed.

Conclusions

MTA Plus mixed with antiwashout gel was found to have lower initial porosity than MTA Plus mixed with water. Both materials exhibited good marginal adaptation and the diffusion of silicon, calcium, and phosphorous across the cement/dentin interface. Thus, the anti–washout-type MTA can be considered to be a suitable substitute for ordinary MTA in all its indications.     

Identification of a Calcium-sensing Receptor in Human Dental Pulp Cells That Regulates Mineral Trioxide Aggregate–induced Mineralization

IntroductionThe purpose of this study was to verify the expression of the calcium-sensing receptor (CaSR) and its role in mineral trioxide aggregate (MTA)-induced odontoblastic differentiation and mineralization in human dental pulp cells (hDPCs).MethodsThe expression of CaSR in human dental pulp tissue and hDPCs was detected using immunohistochemical and immunofluorescent assays. Then, hDPCs were cultured in specific medium supplemented with defined concentrations of MTA dilute alone or in combination with calcimimetic R-568 (a positive allosteric modulator of CaSR [Tocris Bioscience, Bristol, UK]), and cell viability was monitored by Cell Counting Kit-8 (Dojindo Molecular Technologies, Kumamoto, Japan) analysis. Alkaline phosphatase activity, alizarin red S staining, quantitative real-time polymerase chain reaction, and Western blot were used to investigate the gene/protein expression of odontoblastic-associated markers and CaSR in medium supplemented with different combinations of diluted MTA, R-568, and calcilytic Calhex 231 (a negative allosteric modulator of CaSR [Sigma-Aldrich, St Louis, MO]).ResultsCaSR was slightly expressed in the central pulp tissue, whereas it was strongly expressed in the odontoblast layer, plasma membrane, and cytoplasm of hDPCs. Cell Counting Kit-8 assay indicated maximum cell viability in cultures treated with 1:8 diluted MTA additives. Compared with undifferentiated controls, the cells at the early stage of odontoblastic differentiation exhibited lower CaSR protein expression. The combination of 1:8 diluted MTA with 0.1 and 1.0 μmol/L R-568 led to significantly increased cell vitality but decreased alkaline phosphatase activity and mineralized deposit formation, and this negative effect could be attenuated by 1.0 μmol/L Calhex 231 supplementation. Quantitative polymerase chain reaction results showed a significant up-regulation of RUNX2, DSPP, DMP-1, and OCN gene expression in the 1 μmol/L R-568–treated hDPCs. Western blot analysis indicated that the treatment by MTA and R-568 alone or their combination gave no clear trend on the protein levels of CaSR and dentin sialophosphoprotein, whereas Calhex 231 can increase their expressions. In addition, the up-regulation of Akt phosphorylation was observed in R-568– and Calhex 231–treated hDPCs.ConclusionsOur data indicated that CaSR is expressed in human dental pulp and hDPCs and that it can negatively or positively regulate MTA-induced mineralization of hDPCs via the phosphoinositide 3-kinase/Akt pathway in a ligand-dependent manner, suggesting a therapeutic target for modulating reparative dentin formation.     

The Effect of Primary Root Canal Treatment on Dentinal Tubule Penetration of Calcium Silicate–based Sealers during Endodontic Retreatment

IntroductionThe aim of this study was to evaluate the effect of previous root canal treatment on dentinal tubule penetration of calcium silicate–based sealers during endodontic retreatment and to compare 2 different (commonly used and modified) tubule penetration depth measurement techniques.MethodsThe crowns of mandibular premolar teeth were removed, and root canals were prepared (ProTaper Universal; Dentsply Sirona, York, PA) and obturated using AH Plus (AH; Dentsply DeTrey, Konstanz, Germany), MTA Fillapex (MTAF; Angelus, Londrina, Brazil), and WellRoot ST (WRST; Vericom, Gyeonggi-do, South Korea); 0.1% red Rhod-2 dye (Chemodex, St Gallen, Switzerland) was added to the sealers. After storage for 3 weeks (37°C, 100% humid), the following sealer combination subgroups were created (the sealer used during the primary root canal treatment/the sealer used during the retreatment, n = 8): AH/AH, AH/MTAF, and AH/WRST; MTAF/AH, MTAF/MTAF, and MTAF/WRST; and WRST/AH, WRST/MTAF, and WRST/WRST. Green Fluo-3 dye (0.1%) was added to the sealers during retreatment. Sealer penetration depths were measured at 8 points and averaged, and the ratio of the retreatment sealer to the initial sealer was recorded as the mean tubule penetration depth ratio in the first technique, whereas the percentage of the circumferential penetration area ratios of the retreatment and initial sealers was calculated in the second technique. Statistical analyses were performed using Kolmogorov-Smirnov, Kruskal-Wallis, and Mann-Whitney U tests.ResultsThe mean penetration depth of AH and MTFA was found to be higher at the coronal third region in the control groups (P < .05). Both methods showed a significant difference among the groups at the coronal and median regions (P < .05). The AH/WRST, MTAH/AH, MTAF/MTAF, MTAF/WRSTS, and WRST/WRST groups showed deeper tubule penetration (first technique, P = .01). Both techniques showed that the penetration depth of the sealers was the same at the apical region (P > .05), whereas a difference was found among the techniques at the coronal and median regions. The sealers used in the previous root canal treatment limited the penetration depth of the sealers used during the retreatment process (P < .05). When MTAF was used as the initial sealer, the penetration depth of the second sealer did not change, whereas if WRST was the initial sealer, the penetration depth of the sealers at the median and apical regions is negatively affected during the retreatment process (P < .05).ConclusionsPrevious root canal treatment affects the penetration depth of calcium silicate–based sealers. The use of AH Plus and MTA Fillapex during the retreatment process showed an advantage. Both measurement techniques showed similar performances at the apical region, whereas the results varied at the coronal and midcoronal regions; therefore, a more stable sealer penetration measurement technique is still necessary.     

Evaluation of pH and Calcium Ion Release of a Dual-cure Bisphenol A Ethoxylate Dimethacrylate/Mineral Trioxide Aggregate–based Root-end Filling Material

Introduction

The incorporation of light-curable resins has been proposed for mineral trioxide aggregate (MTA) to improve its properties and reduce its setting time. The aim of the present study was to assess the pH and calcium ion release of an experimental bisphenol A ethoxylate dimethacrylate/MTA-based root-end filling material (E-MTA) in comparison with white MTA Angelus (Angelus, Londrina, PR, Brazil) (W-MTA) and to evaluate the influence of the addition of calcium chloride (CaCl₂) on these properties.

Methods

Polyethylene tubes filled with the materials were immersed in deionized water for the measurement of pH (digital pH meter) and calcium release (atomic absorption spectrophotometry). The evaluations were performed at 3 and 24 hours and 7, 15, and 30 days. Data were measured using 2-way repeated measures of variance followed by the Holm-Sidak method (P < .05).

Results

All materials presented a reduction in the pH and released calcium ions during the 30 days of the study. E-MTA showed a significantly lower calcium ion release capacity when compared with W-MTA (P < .05). The calcium release of E-MTA + 5% CaCl₂ was similar to W-MTA (P > .05).

Conclusions

The monomer bisphenol A ethoxylate dimethacrylate added to MTA formed a material with a lower capacity of calcium release than W-MTA despite maintaining a similar pH. However, the addition of CaCl₂ improved the calcium release of this material.     

Initial Transient Accumulation of M2 Macrophage–associated Molecule-expressing Cells after Pulpotomy with Mineral Trioxide Aggregate in Rat Molars

Introduction

M2 (alternatively activated) macrophages are known to participate in wound healing and tissue repair. This study aimed to analyze the temporospatial changes in the distribution and density of M2 macrophage–associated molecule-expressing cells after pulpotomy with mineral trioxide aggregate (MTA) in rat molars to ascertain the role played by M2 macrophages in the healing of MTA-capped pulp tissue.

Methods

The maxillary first molars of 8-week-old Wistar rats were pulpotomized and capped with MTA. After 1–14 days, the teeth were examined after hematoxylin-eosin staining or immunoperoxidase staining of CD68 (a general macrophage marker) and M2 macrophage markers (CD163 and CD204). The density of positively stained cells was enumerated in the surface and inner regions (0–100 μm and 300–400 μm, respectively, from the wound surface).

Results

MTA capping initially caused mild inflammatory changes and the formation of a degenerative layer followed by progressive new matrix formation and calcified bridging. At 1–2 days, CD68-, CD163-, and CD204-positive cells started to accumulate beneath the degenerative layer, and the density of these cells was significantly higher in the surface region than in the inner region (P < .05). From 7 days onward, the 3 types of cells displayed an almost normal distribution beneath the newly formed dentinlike matrix.

Conclusions

After the pulpotomy of rat molars with MTA, M2 macrophage–associated molecule-expressing cells transiently accumulated beneath the degenerative layer under the MTA. This suggests that M2 macrophages participate in the initial phases of the healing of MTA-capped pulp tissue.     

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.     

The Effect of Calcium Hydroxide and Nano–calcium Hydroxide on Microhardness and Superficial Chemical Structure of Root Canal Dentin: An Ex Vivo Study

IntroductionCalcium hydroxide (CH) and nano–calcium hydroxide (NCH) as intracanal medicaments may affect the physical and chemical properties of dentin. The aim of this investigation was to evaluate the effects of CH and NCH on the microhardness and superficial chemical structure of radicular dentin.MethodsIn this in vitro trial, 80 dentin discs were randomly assigned into 2 control and 2 treatment groups (n = 20). CH and NCH pastes were used in the treatment groups. In the control groups, the samples were washed with either normal saline or 2.5% sodium hypochlorite. After 1 and 4 weeks, dentin microhardness was assessed by the Vickers test, and the phosphate/amide I ratio was evaluated by the Fourier-transform infrared spectrometry test. The data were subjected to 1-way and 2-way analyses of variance, the Tukey multiple comparison test, and the Student t test.ResultsA significant reduction in the mean microhardness was observed in the CH group after 4 weeks, whereas the differences were not significant in the NCH and control groups. The amount of reduction induced by NCH was comparable with hypochlorite. A significant increase in the mean phosphate/amide I ratio was observed in the CH and NCH groups compared with the sodium hypochlorite and intact control groups after 1 week, which did not change significantly during the observation period.ConclusionsThe use of CH as an intracanal medicament for 4 weeks reduced dentin microhardness, whereas NCH did not result in any change in the microhardness value. However, a change in the superficial chemical structure was observed earlier after 1 week and in both the CH and NCH groups.     

The Effect of Different Final Irrigant Activation Techniques on the Bond Strength of an Epoxy Resin–based Endodontic Sealer: A Preliminary Study

Introduction

The aim of this study was to evaluate whether or not different final irrigation activation techniques affect the bond strength of an epoxy resin–based endodontic sealer (AH Plus; Dentsply DeTrey, Konstanz, Germany) to the root canal walls of different root thirds.

Methods

Eighty single-rooted human mandibular premolars were prepared by using the ProTaper system (Dentsply Maillefer, Ballaigues, Switzerland) to size F4, and a final irrigation regimen using 3% sodium hypochlorite and 17% EDTA was performed. The specimens were randomly divided into 4 groups (n = 20) according to the final irrigation activation technique used as follows: no activation (control), manual dynamic activation (MDA), CanalBrush (Coltene Whaledent, Altststten, Switzerland) activation, and ultrasonic activation. Five specimens from each group were prepared for scanning electron microscopic observation to assess the smear layer removal after the final irrigation procedures. All remaining roots were then obturated with gutta-percha and AH Plus sealer. A push-out test was used to measure the bond strength between the root canal dentin and AH Plus sealer. The data obtained from the push-out test were analyzed using 2-way analysis of variance and Tukey post hoc tests.

Results

The bond strength values mostly decreased in the coronoapical direction (P < .001). In the coronal and middle thirds, ultrasonic activiation showed a higher bond strength than other groups (P < .05). In the apical third, MDA displayed the highest bond strength to root dentin (P < .05). The majority of specimens exhibited cohesive failures.

Conclusions

The bond strength of AH Plus sealer to root canal dentin may improve with ultrasonic activation in the coronal and middle thirds and MDA in the apical third.