Screw-in force, torque generation, and performance of glide-path files with three rotation kinetics.

We aimed to evaluate the screw-in force, torque generation, and performance of three nickel-titanium (NiTi) glide-path files with different rotational kinetics. ProTaper Ultimate Slider (PULS) and HyFlex EDM Glide-path (HEDG) files were used for canal shaping with constant rotation (CON) or the alternative rotation technique (ART). In the ART mode, the NiTi file was periodically rotated at a speed of 1.5 times faster than that in the CON mode. WaveOne Gold Glider was used with reciprocating motion (WOGG_RCP). Sixty J-shaped resin blocks were assigned to five groups: PULS_CON, PULS_ART, HEDG_CON, HEDG_ART, and WOGG_RCP (n = 12). Glide-path preparation was performed using an automated pecking device. During glide-path preparation, the screw-in force and clockwise and counterclockwise torques were recorded and the number of pecking motions required to reach the working length was determined. The centering ratio was calculated after glide-path preparation using stereomicroscopic images. Data were analyzed using one-way analysis of variance with the Games-Howell post hoc test and the Kruskal-Wallis test with Bonferroni correction. PULS_ART generated a lower maximum screw-in force than PULS_CON. The average number of pecking motions required to reach the working length by HEDG_ART was lower than that by HEDG_CON. The mean centering ratios of PULS_CON and HEDG_CON were - 0.04 and - 0.06, respectively, while those of PULS_ART, HEDG_ART, and WOGG_RCP were 0.09, 0.01, and 0.08, respectively. The ART mode reduced the screw-in force of PULS and enabled faster glide-path preparation with the HEDG file.

Evaluation of design, mechanical properties, and torque/force generation of heat-treated NiTi glide path instruments.

BACKGROUND: Recently, various kinds of heat-treated nickel-titanium (NiTi) glide path instruments have been manufactured. This study aimed to investigate design, phase transformation behavior, mechanical properties of TruNatomy Glider (#17/02), V Taper 2H (#14/03), and HyFlex EDM (#15/03) and compare torque/force generated during simulated glide path preparation with them. METHODS: The designs and phase-transformation behaviors of the instruments were examined via scanning electron microscopy (n = 3) and differential scanning calorimetry (n = 2). Their bending (n = 15), torsional (n = 15), and cyclic fatigue resistances (n = 15) were tested. The ultimate strength and distortion angle were obtained from torsional resistance test. The number of cycles to failure (NCF) was calculated from cyclic fatigue resistance test. The preparation of the glide path was simulated using a double-curved artificial canal (n = 15), and the maximum torque and screw-in forces were measured. Data except NCF was compared between brands with one-way ANOVA with Tukey's honestly significant difference test. NCF was analyzed via Kruskal-Wallis and Mann-Whitney U tests. RESULTS: TruNatomy Glider had the greatest number of threads. TruNatomy Glider showed progressive taper, while V Taper 2H and HyFlex EDM had constant taper. The austenitic transformation-finish temperatures of all the instruments were above body temperature. V Taper 2H demonstrated significantly lower ultimate strength, higher distortion angle, and a higher number of cycles to failure compared with HyFlex EDM and TruNatomy Glider (p < 0.05). The maximum torque generated during preparing glide path was lowest for V Taper 2H, and the maximum screw-in force was lowest for HyFlex EDM (p < 0.05). TruNatomy Glider generated the highest torque and screw-in force during the apical preparation. CONCLUSIONS: V Taper 2H #14/03 showed superior cyclic fatigue resistance and lower ultimate strength. TruNatomy Glider generated greater clockwise torque and screw-in force during apical preparation. The mechanical properties, torque, and screw-force was affected by design of heat-treated glide path instruments. Cervical pre-flaring prior to glide path instrument is recommended.

Morphological measurements of two separate mesiobuccal canals in maxillary first molars using micro-computed tomography.

OBJECTIVES: To obtain radicular measurements of two separate mesiobuccal (MB) root canals in maxillary first molars using micro-computed tomography (µCT) with customized software. MATERIALS AND METHODS: Human maxillary first molar MB roots (N = 36) with two separate canals (MB1, MB2) and apical foramina were scanned by µCT and analyzed with Kappa2 software to reconstruct three-dimensional (3D) surface models of roots and canals. These models were sectioned at 0.1 mm intervals perpendicular to the central axis of each canal. Canal widths, 3D curvatures, and surrounding dentine thicknesses were measured concurrently on each section. Dentine thicknesses were analyzed statistically for differences between each direction and the different levels of both canals. RESULTS: Dentine walls around MB1 were thicker than MB2 (p < 0.05). Thinnest dentine was most often located at disto-inside direction in both canals. Canal widths were significantly smaller in MB2 than MB1 (p < 0.01). Apical constrictions were smaller (p < 0.05) and further (p < 0.05) from the apex in MB2 than MB1. Canal curvatures were greatest in the apical third of both canals (p < 0.001), and they were greater in MB2 than MB1 (p < 0.05). CONCLUSIONS: MB2 canals had shorter lengths, smaller widths, and more severe curvatures and were surrounded by thinner dentine walls. In MB2, apical constrictions were between 1 and 2 mm from the apex, compared to about 1 mm for MB1. CLINICAL RELEVANCE: These detailed measurements and in-depth 3D analyses of maxillary first molar MB roots with two separate canals and apical foramina provide morphologic references for root canal therapy.

Effect of autoclave sterilization on cyclic fatigue and torsional fracture resistance of NiTi rotary instruments.

The purpose of this study is to assess the effect of autoclave sterilization on the cyclic fatigue and torsional fracture resistance of ProTaper Universal (PTU), K3XF, HyFlex EDM (EDM), and TF adaptive (TFA). Sixty instruments from each file type were divided into two categories for cyclic fatigue group (CGr) and torsional fracture group (TGr). CGr and TGr were divided into three subgroups, respectively, consisting of ten instruments from each file type. Cyclic fatigue fracture test was performed using artificial canal made of stainless steel, and the mean number of cycles to failure (NCF) were determined. CGr1, the files were tested to establish baseline for NCF; CGr2, the files were tested cyclic fatigue after 10 cycles of autoclave; CGr3, instruments were autoclaved after being cycled to 25, 50, and 75% of corresponding NCF determined in CGr1, followed by cyclic fatigue test. The torsional fracture test was performed without autoclave (TGr1), after 3-cycle autoclave (TGr2), and 7-cycle autoclave (TGr3), respectively, which evaluated maximum torque and angular deflection. NCF, maximum torque and angular deflection were compared using one-way ANOVA with Bonferroni test. Two-way ANOVA was performed to determine the interaction between 'autoclave treatment' and 'type of NiTi file'. EDM showed highest NCF within the same autoclave treatment. TFA presented the lowest maximum torque and the highest angular deflection, and PTU presented the lowest angular deflection. Within the same NiTi file systems, most of NCF, maximum torque and angular deflection of tested files were not significantly influenced by autoclave condition.

Synthetic Human ß Defensin-3-C15 Peptide in Endodontics: Potential Therapeutic Agent in Streptococcus gordonii Lipoprotein-Stimulated Human Dental Pulp-Derived Cells.

Human ß defensin-3-C15, an epithelium-derived cationic peptide that has antibacterial/antifungal and immuno-regulatory properties, is getting attention as potential therapeutic agent in endodontics. This study aimed to investigate if synthetic human ß defensin-3-C15 (HBD3-C15) peptides could inhibit inflammatory responses in human dental pulp cells (hDPCs), which had been induced by gram-positive endodontic pathogen. hDPC explant cultures were stimulated with Streptococcus gordonii lipoprotein extracts for 24 h to induce expression of pro-inflammatory mediators. The cells were then treated with either HBD3-C15 (50 µg/mL) or calcium hydroxide (CH, 100 µg/mL) as control for seven days, to assess their anti-inflammatory effects. Quantitative RT-PCR analyses and multiplex assays showed that S. gordonii lipoprotein induced the inflammatory reaction in hDPCs. There was a significant reduction of IL-8 and MCP-1 within 24 h of treatment with either CH or HBD3-C15 (p < 0.05), which was sustained over 1 week of treatment. Alleviation of inflammation in both medications was related to COX-2 expression and PGE2 secretion (p < 0.05), rather than TLR2 changes (p > 0.05). These findings demonstrate comparable effects of CH and HDB3-C15 as therapeutic agents for inflamed hDPCs.

Dentinal tubule sealing effects of 532-nm diode-pumped solid-state laser, gallic acid/Fe3+ complex, and three commercial dentin desensitizers.

The purpose of this study was to compare dentinal tubule sealing effects of a 532-nm diode-pumped solid-state (DPSS) laser, gallic acid/Fe3+ complex, and three commercially available dentin desensitizers. Human premolars (n = 44) extracted for orthodontics had standardized cervical cavities prepared, etched (37% phosphoric acid) and randomly assigned to either a control (n = 4), or one of five treatment groups (n = 8/group). Desensitizing treatments were either a 532-nm DPSS laser, gallic acid/Fe3+ complex, oxalate-based Super Seal™ (SS), DIO™ Enamel Coating Pen Pro Tooth (Dio), or adhesive-type Hybrid Coat™ (HC). Dentinal fluid flow (DFF) was monitored continuously in real time during the application of each desensitizing agent, by using a nanoliter-scaled fluid flow-measuring device. Following treatment, morphological changes on dentinal surfaces and within tubules were observed by scanning electron microscopy (SEM). DFF rates were significantly reduced after treatment in all experimental groups (P < 0.05), except SS (P > 0.05). The gallic acid/Fe3+ complex reduced DFF rates the most, and significantly (P < 0.05) more than the three commercial dentin desensitizers. There were no significant differences in DFF reduction rates between the gallic acid/Fe3+ complex and the DPSS laser groups (P > 0.05). There were no significant differences in DFF reduction rates among the three commercial dentin desensitizers (P > 0.05). SEM examination of treated dentin showed that the degree of occlusion of dentinal tubules correlated closely with the corresponding reduction in DFF rates. The gallic acid/Fe3+ complex and 532-nm DPSS laser were superior to other desensitizing methods in occluding dentinal tubules and reducing DFF rates.

Combined effects of mineral trioxide aggregate and human placental extract on rat pulp tissue and growth, differentiation and angiogenesis in human dental pulp cells.

OBJECTIVE: The aim of this study was to evaluate the combined effects of mineral trioxide aggregate (MTA) and human placental extract (HPE) on cell growth, differentiation and in vitro angiogenesis of human dental pulp cells (HDPCs) and to identify underlying signal transduction mechanisms. In vivo dental pulp responses in rats for a pulp-capping agent were examined. MATERIALS AND METHODS: MTS assay. ALP activity test, alizarin red S staining and RT-PCR for marker genes were carried out to evaluate cell growth and differentiation. HUVEC migration, mRNA expression and capillary tube formation were measured to evaluate angiogenesis. Signal transduction was analysed using Western blotting and confocal microscopy. The pulps of rat maxillary first molars were exposed and capped with either MTA or MTA plus HPE. Histologic observation and scoring were performed. RESULTS: Compared to treatment of HDPCs with either HPE or MTA alone, the combination of HPE and MTA increased cell growth, ALP activity, mineralized nodules and expression of marker mRNAs. Combination HPE and MTA increased migration, capillary tube formation and angiogenic gene expression compared with MTA alone. Activation of Akt, mammalian target of rapamycin (mTOR), p38, JNK and ERK MAPK, Akt, and NF-κB were significantly increased by combining HPE and MTA compared with MTA alone. Pulp capping with MTA plus HPE in rats showed superior dentin bridge formation, odontoblastic layers and dentinal tubules and lower inflammatory cell response, compared to the MTA alone group. CONCLUSIONS: This study demonstrates for the first time that the use of MTA with HPE promotes cell growth, differentiation and angiogenesis in HDPCs, which were associated with mTOR, MAPK and NF-κB pathways. Direct pulp capping with HPE plus MTA showed superior results when compared with MTA alone. Thus, the combination of MTA and HPE may be useful for regenerative endodontics.

Comparison of mandibular first molar mesial root canal morphology using micro-computed tomography and clearing technique.

OBJECTIVE: Micro-computed tomography (MCT) with alternative image reformatting techniques shows complex and detailed root canal anatomy. This study compared two-dimensional (2D) and 3D MCT image reformatting with standard tooth clearing for studying mandibular first molar mesial root canal morphology. MATERIALS AND METHODS: Extracted human mandibular first molar mesial roots (n=31) were scanned by MCT (Skyscan 1172). 2D thin-slab minimum intensity projection (TS-MinIP) and 3D volume rendered images were constructed. The same teeth were then processed by clearing and staining. For each root, images obtained from clearing, 2D, 3D and combined 2D and 3D techniques were examined independently by four endodontists and categorized according to Vertucci's classification. Fine anatomical structures such as accessory canals, intercanal communications and loops were also identified. RESULTS: Agreement among the four techniques for Vertucci's classification was 45.2% (14/31). The most frequent were Vertucci's type IV and then type II, although many had complex configurations that were non-classifiable. Generally, complex canal systems were more clearly visible in MCT images than with standard clearing and staining. Fine anatomical structures such as intercanal communications, accessory canals and loops were mostly detected with a combination of 2D TS-MinIP and 3D volume-rendering MCT images. CONCLUSIONS: Canal configurations and fine anatomic structures were more clearly observed in the combined 2D and 3D MCT images than the clearing technique. The frequency of non-classifiable configurations demonstrated the complexity of mandibular first molar mesial root canal anatomy.

A Study on Nanoleakage of Apical Retrograde Filling of Premixed Calcium Silicate-Based Cement Using a Lid Technique.

This study aimed to compare the nanoleakage of retrograde fillings with premixed calcium silicate-based putty and mineral trioxide aggregate (MTA), using two different techniques (traditional and Lid). Sixty-four extracted human teeth were decoronated, then root canals and ends were instrumented for retrograde filling and divided into four groups according to the retrograde filling technique: the traditional and the Lid technique. Each group (n = 15) was filled with Ceraseal + Well-Root putty, Well-Root putty, Ceraseal + ProRoot MTA, and ProRoot MTA. The nanoleakage was evaluated using the Nanoflow device (IB Systems) on days 1, 3, 7, 15 and 30. Data were collected twice per second at the nanoscale (nL/s) and calculated after archiving the stabilization of fluid flow. The Kruskal-Wallis and Mann-Whitney U-tests were used for statistical analysis. All groups showed enhanced sealing ability over time. Regardless of filling materials, the Well-Root putty, Ceraseal+Well-Root putty, and Ceraseal+ProRoot MTA groups indicated less nanoleakage than the ProRoot MTA group in the first week of evaluation (p < 0.05). Although all groups did not show significant differences after 2 weeks, the Ceraseal+ProRoot MTA group leaked less than ProRoot MTA on Days 3 and 7 (p < 0.05). The scanning electron microscopic examined good adaptation to the cavity wall, which was similar to nanoleakage results. Premixed calcium silicate-based putty retrograde filling material alone and using the "lid technique" were shown to be faster and less prone to nanoleakage when compared to MTA.

Growth inhibition and apoptosis-inducing effects of cudraflavone B in human oral cancer cells via MAPK, NF-κB, and SIRT1 signaling pathway.

The goal of this study was to investigate the effect and molecular mechanism of cudraflavone B, a prenylated flavonoid isolated from the root bark of Cudrania tricuspidata, against oral squamous cell carcinoma cells. We observed that cudraflavone B inhibited proliferation of these cells in a time- and dose-dependent manner. At 15 µM, cudraflavone B induced cell death via apoptosis (characterized by the appearance of nuclear morphology) and increased the accumulation of the sub-G1 peak (portion of apoptotic annexin V positive cells). Treatment with cudraflavone B triggered the mitochondrial apoptotic pathway (indicated by induction of the proapoptotic protein p53 and the p21 and p27 effector proteins), downregulation of cell cycle regulatory proteins (e.g., p-Rb, changing Bax/Bcl-2 ratios, cytochrome-c release), and caspase-3 activation. Cudraflavone B time-dependently activated NF-κB, the MAP kinases p38, and ERK, and induced the expression of SIRT1. SIRT1 activator, resveratrol, dose-dependently attenuated the growth-inhibitory and apoptosis-inducing effect of cudraflavone B and blocked cudraflavone B-induced regulatory protein expressions in the mitochondrial pathway such as p53, p21, p27, Bax, caspase-3, and cytochrome-c. Conversely, treatment with SIRT1 inhibitor sirtinol caused opposite effects. These results demonstrate for the first time that the molecular mechanism underlying the antitumor effect in oral squamous cell carcinoma cells is related to the activation of MAPK/and NF-κB as well as of the SIRT1 pathway. Therefore, cudraflavone B may be a lead for the development of a potential candidate for human oral squamous cell carcinoma cells.

A micro-computed tomography study of canal configuration of multiple-canalled mesiobuccal root of maxillary first molar.

OBJECTIVES: Detailed information of complex anatomical configuration of mesiobuccal (MB) root is essential for successful endodontic treatment in maxillary first molars. The aims of this study were to investigate the configuration types present in multiple-canalled MB roots of maxillary first molars using micro-computed tomography (µCT) and to evaluate whether further modification to current configuration classifications are needed for in-depth morphology study of MB root canal system. MATERIALS AND METHODS: One hundred and fifty-four extracted human maxillary first molar MB roots were scanned by µCT (Skyscan) and their canals were reconstructed by 3D modeling software. Root canal configurations were categorized according to the classifications proposed by Weine and Vertucci. Canal configurations that did not fit into both classifications were categorized as non-classifiable. RESULTS: One hundred and thirteen (73.4 %) MB roots had multiple canals. The most predominant canal configuration was Weine type III (two orifices and two foramens). Thirty-three (29.2 %) and 20 (17.7 %) MB roots had non-classifiable configuration types that could not be classified by the Weine and Vertucci classification, respectively. Three configurations (types 1-3, 2-3-2-3-2, and 2-3-4-3-2) were first reported in maxillary first molar MB roots. CONCLUSIONS: The present µCT study provided an in-depth analysis of canal configurations of the MB roots of maxillary first molar and suggests that additional modification of current configuration classifications may be needed to more accurately reflect the morphology configurations of MB roots. CLINICAL RELEVANCE: Clinicians should consider the complex canal configurations of the maxillary first molar MB roots during surgical or nonsurgical endodontic procedures.

Effect of heat treatment on cyclic fatigue resistance, thermal behavior and microstructures of K3 NiTi rotary instruments.

OBJECTIVE: The aim of this study was to investigate the effect of heat treatment on the cyclic fatigue resistance, thermal behavior and microstructural changes of K3 NiTi rotary instruments. MATERIALS AND METHODS: Twelve control (as-received) and 12 experimental (heat-treated) K3 NiTi rotary instruments were compared in this study. Those experimental K3 instruments were heated in a furnace for 30 min at 450°C and then quenched in water. The cyclic fatigue resistance was measured with a fatigue tester. The thermal characteristic and the microstructures of both instruments were investigated by differential scanning calorimetry (DSC) and transmission electron microscopy (TEM), respectively. RESULTS: There was a significant increase in the cyclic fatigue resistance between the heat-treated instruments and the as-received instruments (T-test, p < 0.05). DSC showed that the as-received and heat-treated samples were different, with an increased Af (austenite-finish temperature) for the latter. TEM analysis revealed that both as-received and heat-treated instruments were composed mainly of an austenite phase. However, the heat-treated samples had an increased appearance of larger grains, twinning martensite, TiO2 surface layer and a Ni-rich inner layer. CONCLUSIONS: Heat treatment increased the cyclic fatigue resistance of NiTi files and changed the thermal behavior of the instruments without marked changes in the constituting phases of NiTi alloy.

Buckling resistance, torque, and force generation during retreatment with D-RaCe, HyFlex Remover, and Mtwo retreatment files.

Objectives: This study compared the buckling resistance of 3 nickel-titanium (NiTi) retreatment file systems and the torque/force generated during retreatment. Materials and Methods: The buckling resistance was compared among the D-RaCe (DR2), HyFlex Remover, and Mtwo R25/05 retreatment systems. J-shaped canals within resin blocks were prepared with ProTaper NEXT X3 and obturated by the single-cone technique with AH Plus. After 4 weeks, 4 mm of gutta-percha in the coronal aspect was removed with Gates-Glidden drills. Retreatment was then performed using DR1 (size 30, 10% taper) followed by DR2 (size 25, 4% taper), HyFlex Remover (size 30, 7% taper), or Mtrwo R25/05 (size 25, 5% taper) (15 specimens in each group). Further apical preparation was performed with WaveOne Gold Primary. The clockwise torque and upward force generated during retreatment were recorded. After retreatment, resin blocks were examined using stereomicroscopy, and the percentage of residual filling material in the canal area was calculated. Data were analyzed using 1-way analysis of variance with the Tukey test. Results: The HyFlex Remover files exhibited the greatest buckling resistance (p < 0.05), followed by the Mtwo R25/05. The HyFlex Remover and Mtwo R25/05 files generated the highest maximum clockwise torque and upward force, respectively (p < 0.05). The DR1 and DR2 files generated the least upward force and torque (p < 0.05). The percentage of residual filling material after retreatment was not significantly different between file systems (p > 0.05). Conclusions: NiTi retreatment instruments with higher buckling resistance generated greater clockwise torque and upward force.

Physicochemical and biological properties of four calcium silicate-based endodontic cements.

Background/purpose: Several brands of calcium silicate-based cements (CSBCs) are currently marketed. Here we compared physicochemical and biological properties of new products Ortho MTA (BioMTA), Retro MTA (BioMTA), and EZ-Seal (Ezekiel) to widely used ProRoot MTA (Dentsply Tulsa). Materials and methods: CSBCs were analyzed by X-ray diffractometry and examined by scanning electron microscopy. Elemental composition was determined by energy dispersive spectroscopy. Particle size was measured by particle size analyzer. Human stem cells from apical papilla (SCAPs) were incubated with eluates from CSBCs. Survival of SCAP cells was evaluated with MTT assay. The Alizarin red S stain was used to identify calcified nodules formed in SCAP cultures. The effects of CSBC eluates on SCAP proliferation and migration were examined using an in-vitro scratch "wound-healing" assay. Results: All CSBC specimens showed similar X-ray diffraction patterns. The average particle size of EZ-Seal was smaller than ProRoot MTA, Ortho MTA, and Retro MTA (P < 0.001). The least cytotoxicity of eluates was found for EZ-Seal. In the Alizarin red S staining test, calcified nodules were observed in cultures with ProRoot MTA, Ortho MTA, and Retro MTA, however, no calcified nodules were observed in cultures with EZ-Seal. SCAP proliferation and migration capacity in presence of EZ-Seal was higher than with ProRoot MTA, Ortho MTA, and Retro MTA (P < 0.001). Conclusion: EZ-Seal has a smaller average particle size and a better cytocompatibility than all other examined CSBCs.

Effect of bioactive glass addition on the physical properties of mineral trioxide aggregate.

BACKGROUND: The addition of bioactive glass (BG), a highly bioactive material with remineralization potential, might improve the drawback of weakening property of mineral trioxide aggregates (MTA) when it encounters with body fluid. This study aims to evaluate the effect of BG addition on physical properties of MTA. METHODS: ProRoot (MTA), and MTA with various concentrations of BG (1, 2, 5 and 10% BG/MTA) were prepared. Simulated body fluid (SBF) was used to investigate the effect of the storage solution on dentin remineralization. Prepared specimens were examined as following; the push-out bond strength to dentin, compressive strength, setting time solubility and X-ray diffraction (XRD) analysis. RESULTS: The 2% BG/MTA showed higher push-out bond strengths than control group after 7 days of SBF storage. The 2% BG/MTA exhibited the highest compressive strength. Setting times were reduced in the 1 and 2% BG/MTA groups, and solubility of all experimental groups were clinically acceptable. In all groups, precipitates were observed in dentinal tubules via SEM. XRD showed the increased hydroxyapatite peaks in the 2, 5 and 10% BG/MTA groups. CONCLUSION: It was verified that the BG-added MTA increased dentin push-out bond strength and compressive strength under SBF storage. The addition of BG did not negatively affect the MTA maturation reaction; it increased the amount of hydroxyapatite during SBF maturation.

Characterization of Histopathology and Microbiota in Contemporary Regenerative Endodontic Procedures: Still Coming up Short.

INTRODUCTION: This study aimed to investigate microbiota and the histopathology of infected immature teeth microenvironments after disinfection with calcium hydroxide, triple antibiotic paste, and a synthetic antimicrobial peptide (synthetic human beta-defensin-3-C15) for regenerative endodontic procedures (REPs). The null hypothesis was that there is no difference among intracanal medications on disinfection in REPs. METHODS: Pulp necrosis and periapical lesions were induced in immature beagle dog premolars. Block randomized teeth were uninfected (negative control, n = 6), left infected (positive control, n = 6), or medicated with a disinfectant (n = 6/group). After disinfection (2 weeks), teeth were reaccessed, irrigated with 17% EDTA, blood clot induced, sealed with ProRoot MTA (Dentsply Tulsa Dental, Tulsa, OK), and restored with resin-modified glass ionomer. Animals were monitored radiographically and euthanized (12 weeks) for histopathologic and metagenomic analyses. RESULTS: REP-treated roots showed radiographic repair of periapical radiolucency (67.65%, 23/34), continued root development (73.53%, 25/34), and apical closure (70.59%, 24/34) regardless of the disinfectant used (P > .05). Canal microenvironments histologically devoid of bacteria contained new mineralized and pulp-like tissues in characteristic patterns that varied by disinfectant. Next-generation sequencing (16S ribosomal RNA) identified Firmicutes, Proteobacteria, Actinobacteria, and Bacteroidetes as dominant phyla of microbiota in immature teeth. Infection-induced teeth showed changes in diversity and richness of microbiota from negative controls. Compared with positive controls, all treated teeth exhibited depleted operational taxonomic units, with lower phylogenic diversity from synthetic human beta-defensin-3-C15-treated teeth. CONCLUSIONS: There were no differences among the medicaments investigated in radiologic treatment outcomes, but disinfectants in REPs showed altered microbiota from normal and diseased immature teeth with different histologic patterns of regeneration.

Effects of blood contamination on microtensile bond strength to dentin of three self-etch adhesives.

This study evaluated the effects of blood contamination and decontamination methods during different steps of bonding procedures on the microtensile bond strength of two-step self-etch adhesives to dentin. Sixty extracted human molars were ground flat to expose occlusal dentin. The 60 molars were randomly assigned to three groups, each treated with a different two-step self-etch adhesive: Clearfil SE Bond, AdheSE and Tyrian SPE. In turn, these groups were subdivided into five subgroups (n = 20), each treated using different experimental conditions as follows: control group-no contamination; contamination group 1-CG1: primer application/ contamination/primer re-application; contamination group 2-CG2: primer application/contamination/wash/dry/primer re-application; contamination group 3-CG3: primer application/adhesive application/light curing/contamination/ adhesive re-application/light curing; contamina- tion group 4-CG4: primer application/adhesive application/light curing/contamination/wash/ dry/adhesive re-application/light curing. Composite buildup was performed using Z250. After 24 hours of storage in distilled water at 37 degrees C, the bonded specimens were trimmed to an hourglass shape and serially sectioned into slabs with 0.6 mm2 cross-sectional areas. Microtensile bond strengths (MTBS) were assessed for each specimen using a universal testing machine. The data were analyzed by two-way ANOVA followed by a post hoc LSD test. SEM evaluations of the fracture modes were also performed. The contaminated specimens showed lower bond strengths than specimens in the control group (p < 0.05), with the exception of CG1 in the Clearfil SE group and CG2 and CG3 in the Tyrian SPE group. Among the three self-etch adhesives, the Tyrian SPE group exhibited a significantly lower average MTBS compared to the Clearfil SE Bond and AdheSE (p < 0.05) groups. Based on the results of the current study, it was found that blood contamination reduced the MTBS of all three self-etch adhesives to dentin, and water-rinsing was unable to overcome the effects of blood contamination.

Sealing Ability of AH Plus and GuttaFlow Bioseal.

The objective of root canal obturation is to achieve a fluid-tight seal. Recently, GuttaFlow bioseal (GB), a root canal sealer composed of polydimethylsiloxane, gutta-percha particles, and bioactive glass ceramics, has been developed, to enhance the sealing ability of root canal filling material. The objective of this study was to assess the sealing ability of GB using a subnanoliter-scaled fluid-flow measuring device and to compare with that of AH Plus (AH). The fluid flow in root canal-filled teeth using either gutta-percha cone (GP) with AH (GAR; n = 10) or GP with GB (GBR; n = 10) and in GP inserted in AH blocks (GA; n = 10) or GP inserted in GB blocks (GB; n = 10) was measured. In addition, fluid flow in sealer blocks of AH (A; n = 10) and GB (B; n = 10), which served as negative controls, was measured. Root canal-filled teeth using GP without any sealer (GR) acted as positive controls (n = 10). The leakage was obtained by calculating the volume of moved water by time (s), after stabilization of the fluid flow was achieved. Statistical analysis was performed using the Kruskal-Wallis test and Mann-Whitney U-test with Bonferroni correction. A p value less than 0.00238 (0.05/21) was considered significantly different. The mean leakages (nL/s) in the groups are as follows: GAR, 0.0958 ± 0.0543; GBR, 0.0223 ± 0.0246; GA, 0.0644 ± 0.0803; GB, 0.0267 ± 0.0182; A, 0.0055 ± 0.0057; B, 0.0052 ± 0.005; and GR, 0.2892 ± 0.3018. The mean leakage in the GBR group was lower than that in the GAR group (p = 0.001), while the mean leakages in the GA and GB groups were not significantly different. GuttaFlow bioseal can be useful in single-cone obturation technique.

Bending resistance and cyclic fatigue resistance of WaveOne Gold, Reciproc Blue, and HyFlex EDM instruments.

BACKGROUND/PURPOSE: Several single-file systems manufactured using different heat treatment and operated by different kinematics have been released. This study compared the bending resistance and cyclic fatigue resistance of three NiTi files, and examined their phase-transformation behavior. MATERIALS AND METHODS: WaveOne Gold Primary (WOG), Reciproc Blue R25 (RPB), and HyFlex EDM OneFile (HDM) were tested (N = 40/instrument). A vertical load was applied to a point 3 mm from the tip, and the stress was measured until a displacement of 3 mm. Tests were conducted at either room temperature (RT: 22 °C) or body temperature (BT: 37 °C) (n = 10). Cyclic fatigue resistance tests were performed in an artificial canal, which had a curvature angle of 40° and a 5-mm radius. Tests were conducted at either RT or BT (n = 10). Instruments were operated according to the manufacturers' instructions. Test results were analyzed using the Kruskal-Wallis and the Mann-Whitney tests. Additional three instruments of each brand were subjected to differential scanning calorimetry (DSC). RESULTS: At RT the bending resistance of three files were not significantly different. However, at BT the bending resistance of RPB was highest, followed by WOG, and HDM (P < 0.05). At RT, RPB demonstrated the longest fracture time, followed by HDM, and WOG (P < 0.05). At BT, HDM had the longest fracture time, followed by RPB, and WOG (P < 0.05). The WOG, RPB consisted of austenite in a considerable proportion, whereas HDM was mainly martensite state at BT. CONCLUSION: HDM presented superior flexibility and cyclic fatigue resistance at BT.

In Vitro Characterization of Dental Pulp Stem Cells Cultured in Two Microsphere-Forming Culture Plates.

Various three-dimensional (3D) culture methods have been introduced to overcome the limitations of in vitro culture and mimic in vivo conditions. This study aimed to evaluate two microsphere-forming culture methods and a monolayer culture method. We evaluated cell morphology, viability, osteo-, adipo-, and chondrogenic differentiation potential of dental pulp stem cells (DPSCs) cultured in 3D culture plates: ultra-low attachment (ULA) and U-bottomed StemFit 3D (SF) plates, and a two-dimensional (2D) monolayer plate. RNA sequencing (RNA-seq) revealed differentially expressed gene (DEG) profiles of the DPSCs. In contrast to an increasing pattern in the 2D group, cell viability in 3D groups (ULA and SF) showed a decreasing pattern; however, high multilineage differentiation was observed in both the 3D groups. RNA-seq showed significantly overexpressed gene ontology categories including angiogenesis, cell migration, differentiation, and proliferation in the 3D groups. Hierarchical clustering analysis revealed a similar DEG regulation pattern between the 3D groups; however, a comparatively different DEG was observed between the 2D and 3D groups. Taken together, this study shows that DPSCs cultured in microsphere-forming plates present superior multilineage differentiation capacities and demonstrate higher DEG expression in regeneration-related gene categories compared to that in DPSCs cultured in a conventional monolayer plate.