Penetration of Calcium Silicate and Epoxy Resin Sealers Into the Lateral Canals.

OBJECTIVES: The aim of this research was to compare the penetration ability of calcium silicate-based sealers (iRoot SP and TotalFill BC HiFlow) and an epoxy resin-based sealer (AH Plus) into the lateral canals using the single-cone (SC) or continuous wave compaction (CW) obturation techniques. METHODS: Ninety single-rooted human teeth received canal instrumentation and diaphanisation before artificial lateral canals were created at 3 different root levels. The specimens were randomly allocated into 6 groups based on the combination of obturation technique and sealer used. Radiographic and photographic measurements evaluated the percentage of sealer penetration. Statistical analysis was performed to compare the penetration ability amongst different types of sealers, obturation methods, and root levels. RESULTS: Although TotalFill BC HiFlow sealer combined with the CW technique demonstrated greater sealer penetration at the coronal level (P < .05), the overall penetration ability of iRoot SP, TotalFill BC HiFlow, and AH Plus was not significantly different. The deepest sealer penetration was observed at the apical root level. CONCLUSIONS: All sealers showed similar penetration abilities into the lateral canals using the SC or CW techniques in vitro. Calcium silicate-based sealers have comparable penetration ability into lateral canals compared to epoxy resin-based sealers using SC or CW obturation techniques.

Reliability and reproducibility of CBCT assessment of mandibular changes before and after treatment for Class III growing patients - an easy and quick way for evaluation.

The objective of this study was to evaluate intraobserver reliability and inter-observer reproducibility of a 3-dimensional (3D) assessment method for mandibular changes of growing patients after orthodontic treatment for Class III malocclusion.Methods Cone-beam computed tomography (CBCT) scans were performed before and after orthodontic treatment for 27 patients. During the scan, the patient was positioned such that his/her mandibular plane was parallel to floor. Three observers independently worked on the DICOM data, reconstructed the pre- and post-treatment 3D models in software, selected the stable anatomical structures (basal bone area from the lingual surface of the symphysis to the distal aspect of the first molars) to guide the automated superimposition process. Then, each observer registered 14 anatomical landmarks on the virtual models, for three times after suitable interval, to generate 3 sets of coordinates; the mean was taken as the coordinates for that particular landmark. The intraobserver reliability and inter-observer reproducibility of the method were analyzed.Results The ICCs was > 0.90 for 25 (92.6%) of the intraobserver assessments. The precision of the measurement method was < 0.3 mm in 24 (88.9%) cases. The interobserver reproducibility errors were < 0.3 mm in 21 of the 27 cases.Conclusions The intraobserver reliability and inter-observer reproducibility of 3D assessment of mandibular changes using the virtual models were excellent.

Tissue Stabilization, Bacterial Adhesion, and Stem Cell Viability in Trans-cinnamaldehyde-conditioned Dentin.

INTRODUCTION: This laboratory study aimed to evaluate the effect of trans-cinnamaldehyde (TC) conditioning on dentin tissue stabilization, bacterial adhesion, and stem cell toxicity. METHODS: Dentin beams (n = 204) from extracted human molars were demineralized in phosphoric acid and treated with TC (2.5, 5, and 7.5%), 50% ethanol-water mixture (vehicle control) or 2.5% glutaraldehyde (GA) (positive control) for 30 minutes. Demineralized but untreated specimens served as the negative control. After treatment, collagen crosslinking was characterized by measuring the elastic modulus (Er) and hardness (n = 5). Biodegradation resistance was examined by determining the loss of dry mass (n = 8), hydroxyproline release (n = 4) and scanning electron microscopy (n = 2), after exposure to bacterial collagenase. Inhibition of bacterial adhesion was investigated by colony counting assay (n = 12) and scanning electron microscopy (n = 2). Viability of stem cells of the apical papilla on TC-conditioned dentin was determined using the Cell Counting Kit-8 assay (n = 8). Data were statistically analyzed using one-way analysis of variance (ANOVA) test followed by Dunnett's multiple comparisons at a significance level of 5%. RESULTS: TC-conditioned dentin showed a concentration-dependent increase in Er and hardness. The Er and hardness of 5% and 7.5% TC-conditioned dentin were significantly greater than that of the negative control and vehicle control groups (P < .05). There was no significant difference in the biodegradation resistance between GA and 5% TC-conditioned dentin (P > .05). TC-conditioned dentin showed a well-preserved collagen fibril network with clear cross-banding, comparable to GA-conditioned dentin. All concentrations of TC inhibited bacterial adhesion on dentin, significantly greater than the negative control (P < .05). There was no reduction in viability of stem cells of the apical papilla viability on TC-conditioned dentin compared to the negative control (P > .05). CONCLUSIONS: TC conditioning stabilized the dentin and protected it from enzymatic degradation. TC prevented bacterial adhesion on the dentin but maintained stem cell viability.

Intracellular bacterial eradication using a novel peptide in vitro.

AIM: To determine the effect of a novel antimicrobial peptide (AMP; OP145) and cell-penetrating peptide (Octa-arginine/R8) conjugate on the killing of intracellular Enterococcus faecalis, compared to OP145 and an antibiotic combination recommended for regenerative endodontic procedures. METHODOLOGY: The biocompatible concentrations of OP145 and OP145-R8 were determined by assessing their cytotoxicity against human macrophages and red blood cells. Spatiotemporal internalization of the peptides into macrophages was investigated qualitatively and quantitatively by confocal laser scanning microscopy and flow cytometry respectively. Killing of extracellular and intracellular E. faecalis OG1RF by the peptides was determined by counting the colony-forming units (CFU). Intracellular antibacterial activity of the peptides was compared to a double antibiotic combination. Confocal microscopy was used to confirm the intracellular bacterial eradication. Significant differences between the different test groups were analysed using one-way analysis of variance. p < .05 was considered to be statistically significant. RESULTS: Peptides at a concentration of 7.5 µmol/L were chosen for subsequent experiments based on the results of the alamarBlue™ cell viability assay and haemolytic assay. OP145-R8 selectively internalized into lysosomal compartments and the cytosol of macrophages. Conjugation with R8 improved the internalization of OP145 into macrophages in a temporal manner (70.53% at 1 h to 77.13% at 2 h), while no temporal increase was observed for OP145 alone (60.53% at 1 h with no increase at 2 h). OP145-R8 demonstrated significantly greater extracellular and intracellular antibacterial activity compared to OP145 at all investigated time-points and concentrations (p < .05). OP145-R8 at 7.5 µmol/L eradicated intracellular E. faecalis after 2 h (3.5 log reduction compared to the control; p < .05), while the antibiotics could not reduce more than 0.5 log CFU compared to the control (p > .05). Confocal microscopy showed complete absence of E. faecalis within the OP145-R8 treated macrophages. CONCLUSIONS: The results of this study demonstrated that the conjugation of an AMP OP145 to a cell-penetrating peptide R8 eradicated extracellular and intracellular E. faecalis OG1RF without toxic effects on the host cells.

Long-term follow-up of single-visit pulp revascularisation for the dens evaginatus and dens invaginatus: Cases report.

This article documents cases of single-visit pulp revascularisation for dens evaginatus and dens invaginatus, without using intracranial medicaments or antibiotics, aiming to provide a potentially applicable protocol for pulp revascularisation procedure in a single-visit. Two patients with chief complaints of pain and swelling visited a dental hospital. Radiographs revealed that the causative teeth had an open apex and periapical radiolucency, and the teeth were diagnosed as pulp necrosis and acute apical abscess or symptomatic apical periodontitis. For both cases, single-visit revascularisation was completed without intracanal medicaments or antibiotics. The patients were periodically recalled to evaluate periapical healing after treatment. The apical lesion healed, and the root dentin thickening was observed. The single-visit pulp revascularisation procedure without using specific intracanal medicaments can produce clinically favourable results for these dental anomalies.

Effect of MMP Inhibitors on the Bond Strength of Fibre Posts After Ageing.

AIM: This in vitro study aimed to evaluate the effect of matrix metalloproteinase (MMP) inhibitors on the bond strength of resin-cemented fibre posts to radicular dentin under an aged-loaded condition. MATERIALS AND METHODS: Radicular dentin was prepared and irrigated by MMP inhibitor solution after root canal obturation in 60 extracted single-rooted teeth based on 6 groups: (1) 2% chlorhexidine (CHX) + loaded; (2) CHX + unloaded; (3) 0.5% benzalkonium chloride (BAC) + loaded; (4) BAC + unloaded; (5) 17% ethylenediaminetetraacetic acid (EDTA) + loaded; and (6) EDTA + unloaded. After final rinsing, all specimens were sliced cross-sectionally and kept in a water bath for 12 months of ageing. Groups 1, 3, and 5 were subjected to cyclic loading. Push-out tests were conducted using a universal testing machine, and failure mode was examined. The data were analysed using 3-way analysis of variance and post hoc tests at α = 0.05. RESULTS: BAC + unloaded demonstrated the highest mean bond strength (3.12 ± 0.18 MPa; P < .001), while the BAC + loaded and CHX + loaded groups showed a significantly lower push-out bond strength than their unloaded counterparts. Mixed adhesive-cohesive failure was the most common failure mode observed. CONCLUSIONS: Without cycling loading, BAC was superior to CHX and EDTA in preserving the bond strength of resin-cemented fibre posts after 12 months of ageing. Loading significantly weakened the effectiveness of BAC and CHX in preserving the bond strength.

PROMs Following Root Canal Treatment and Surgical Endodontic Treatment.

The FDI is currently working on developing a tool to encompass patient-reported outcome measures (PROMs) within the overall assessment of outcomes of endodontic treatment. The outcome of endodontic treatment has traditionally been determined by various clinical and radiographic criteria. However, these parameters do not address the impact of treatment on a patient's oral health-related quality of life (OHRQoL). OHRQoL, a crucial PROM, can be used to understand treatment outcome from a patient-centred perspective, thus improving clinician-patient communication whilst guiding decision-making. This focussed review aims to recount the OHRQoL of patients following nonsurgical root canal treatment and surgical endodontic treatment, with a specific focus on the minimal important difference (MID; the minimum score changes of an outcome instrument for a patient to register a clinically significant change in their OHRQoL and/or oral condition) and the methods used to determine it. The current evidence indicates that the OHRQoL of patients requiring root canal treatment is poorer than those without such need. Accordingly, the literature suggests that OHRQoL improves following nonsurgical or surgical endodontic treatment. However, study methodologies vary widely, and conclusions cannot be drawn with high confidence, nor can MID recommendations be provided. Well-designed clinical studies with baseline measurements and appropriate follow-up time frames are therefore needed. Despite that the literature is rife with outcome studies, research on PROMs is an area that deserves greater attention, particularly in relation to the MID. Determining the MID will facilitate the understanding of changes in outcome scores from the patients' perspective, thus allowing for more informed decision-making in clinical practice.

A simple AI-enabled method for quantifying bacterial adhesion on dental materials.

Purpose: Measurement of bacterial adhesion has been of great interest for different dental materials. Various methods have been used for bacterial counting; however, they are all indirect measurements with estimated results and therefore cannot truly reflect the adhesion status. This study provides a new direct measurement approach by using a simple artificial intelligence (AI) method to quantify the initial bacterial adhesion on different dental materials using Scanning Electron Microscope (SEM) images. Materials and Methods: Porphyromonas gingivalis (P.g.) and Fusobacterium nucleatum (F.n.) were used for bacterial adhesion on dental zirconia surfaces, and the adhesion was evaluated using SEM images at time points of one, seven, and 24 h(s). Image pre-processing and bacterial area measurement were performed using Fiji software with a machine learning plugin. The same AI method was also applied on SEM with Streptococcus mutans (S.m.) inoculated PMMA nano-structured surface at 1, 24, 72, and 168 h(s), and then further compared with the CLSM method. Results: For both P.g. and F.n. initiation adhesion on zirconia, a new linear correlation (r2 > 0.98) was found between bacteria adhered area and time, such that: b acteria   adhered   area   ( m m 2 ) ∝ log ⁡ ( time ) For S.m. on PMMA surface, live/dead staining CLSM method and the newly proposed AI method on SEM images were strongly and positively associated (Pearson's correlation coefficient r > 0.9), i.e. both methods are comparable. Conclusions: SEM images can be analyzed directly for both morphology and quantifying bacterial adhesion on different dental materials' surfaces by the simple AI-enabled method with reduced time, cost, and labours.

Enterococcus faecalis Shields Porphyromonas gingivalis in Dual-Species Biofilm in Oxic Condition.

Aim: To develop a reproducible biofilm model consisting of Enterococcus faecalis (E. faecalis) and Porphyromonas gingivalis (P. gingivalis) and to evaluate the interaction between the two bacterial species. Methodology: E. faecalis and P. gingivalis were grown in mono-culture, sequential, and co-culture models for 96 h in a 96-well polystyrene microtiter plate under both aerobic and anaerobic conditions separately. The viability of the two bacterial species in the biofilms was quantified by polymerase chain reaction (qPCR). Biofilm thickness and protein contents were measured using confocal laser scanning microscopy (CLSM). Two-way analysis of variance (ANOVA) was performed to analyze cell viability and biofilm thickness among different culture models cultivated under either aerobic or anaerobic conditions. The level of significance was set at p < 0.05. Results: Different culture models tested did not show any significant difference between the viable cell counts of both E. faecalis and P. gingivalis cultivated under aerobic and anaerobic conditions (p > 0.05). Biofilm was significantly thicker (p < 0.05) in the co-culture models compared to the mono-culture and sequential models. Protein contents in the biofilms were more pronounced when both bacterial species were co-cultured under aerobic conditions. Conclusions: E. faecalis appeared to shield P. gingivalis and support its continued growth in oxic (aerobic) conditions. The co-culture model of E. faecalis and P. gingivalis produced a significantly thicker biofilm irrespective of the presence or absence of oxygen, while increased protein contents were only observed in the presence of oxygen.

Clinical efficacy of activated irrigation in endodontics: a focused review.

Root canal debridement, which includes the removal of infected tissues and microbial biofilms, is considered the corner stone of root canal treatment. Chemical adjuncts play a multitude of functions in this regard, as tissue solvents, antimicrobial agents and for removing the smear layer. These adjuncts (irrigants) are usually delivered using a syringe and needle. With increasing knowledge of the complexity of root canal anatomy and tenacity of microbial biofilms, the need for strategies that potentiate the action of these irrigants within the root canal system cannot be overemphasized. Several such activated irrigation strategies exist. The aim of this review is to comprehensively discuss the different irrigant activation methods from the context of clinical studies.

A nanostructured anti-biofilm surface widens the efficacy against spindle-shaped and chain-forming rod-like bacteria.

Current control of pathogenic bacteria at all biomaterial interfaces is poorly attuned to a broad range of disease-causing pathogens. Leading antimicrobial surface functionalization strategies with antimicrobial peptides (AMPs), defensins, have not shown their promised efficacy. One of the main problems is the lack of stability and swift clearance from the surface. Surface nanotopography bearing sharp protrusions is a non-chemical solution that is intrinsically stable and long-lasting. Previously, the geometrically ordered arrays of nanotipped spines repelled or rapidly ruptured bacteria that come into contact. The killing properties so far work on cocci and rod-like bacteria, but there is no validation of the efficacy of protrusional surfaces on pathogenic bacteria with different sizes and morphologies, thus broadening the utility of such surfaces to cover increasingly more disease entities. Here, we report a synthetic analogue of nanotipped spines with a pyramidal shape that show great effectiveness on species of bacteria with strongly contrasting shapes and sizes. To highlight this phenomenon in the field of dental applications where selective bacterial control is vital to the clinical success of biomaterial functions, we modified the poly(methyl)-methacrylate (PMMA) texture and tested it against Streptococcus mutans, Enterococcus faecalis, Porphyromonas gingivalis, and Fusobacterium nucleatum. These nanopyramids performed effectively at levels well above those of normal and roughened PMMA biomaterials for dentistry and a model material for general use in medicine and disease transmission in hospital environments.

Comparison of In Vitro Torque Generation during Instrumentation with Adaptive Versus Continuous Movement.

INTRODUCTION: The purpose of this in vitro study was to compare the torque generated by continuous and adaptive movements of 2 nickel-titanium rotary file systems. METHODS: Forty-five simulated resin blocks with an S-shaped canal were randomly divided into 3 groups (n = 15) according to the file system and kinematics: the K3XF rotary system (Kerr Endodontics, Orange, CA) with continuous rotary movement, the K3XF with adaptive movement, and the Twisted File (Kerr Endodontics) adaptive file with adaptive movement. After creating a glide path, the canal was instrumented to the same size (.04/#20 for K3XF or SM1 for the Twisted File with adaptive movement) before torque measurement. During the final instrumentation procedure with the .06/25 sized file (K3XF or SM2), the generated torque and the preparation time were recorded. The total torque experienced and the maximum torque value were calculated. The data were statistically analyzed using 1-way analysis of variance and the Tukey post hoc comparison test at a significance level of 95%. RESULTS: The K3XF file system used with adaptive motion group showed significantly lower total and maximum torque values compared with the K3XF with continuous rotary movement group. The Twisted File adaptive file with adaptive motion showed significantly lower torque generation and shorter preparation time than the K3XF groups with adaptive or continuous rotation (P < .05). CONCLUSIONS: Under the conditions of this study, adaptive movement for nickel-titanium files may reduce torque generation without increasing preparation time. Nickel-titanium files with a smaller cross-sectional area using adaptive movement may be helpful to reduce the potential risk of root dentin damage.

Effect of instrumentation systems on endotoxin reduction from root canal systems: A systematic review of clinical studies and meta-analysis.

This systematic review assessed the effect of different root canal instrumentation systems on endotoxin reduction from the root canal system. The literature search was conducted in two electronic databases (PubMed and Scopus) using specific key words. The search strategy followed the PRISMA guidelines. Qualitative synthesis and quantitative synthesis of the data were performed and data interpretation was done based on the guidelines in the Cochrane Handbook. The risk of bias was assessed using Cochrane criteria. The initial search yielded 600 citations, of which three papers met the criteria for inclusion in this review. Studies showed a significant reduction in endotoxin following rotary and reciprocating instrumentation, with no significant differences between them. The meta-analysis showed no statistical significant difference between reciprocation and rotary files (P > 0.05). In conclusion, the instrumentation techniques compared in this review decrease endotoxin content from root canals, with no significant difference between them.

Effect of the Glide Path Establishment on the Torque Generation to the Files during Instrumentation: An In Vitro Measurement.

INTRODUCTION: The purpose of this study was to compare in vitro torque generation during instrumentation with or without glide path establishment. METHODS: Endo-training resin blocks with J-shaped canals were randomly divided into 2 groups according to glide path establishment (with or without) and subdivided into 2 subgroups with shaping instruments (WaveOne [Dentsply Maillefer, Ballaigues, Switzerland] or WaveOne Gold [Dentsply Maillefer]) (n = 15). For the glide path-established group, the glide path was prepared using ProGlider (Dentsply Maillefer). During the instrumentation with WaveOne or WaveOne Gold, in vitro torque was measured. The acquired data were analyzed with software. The maximum torque and total torque (the sum of the generated torque) were calculated. The data were statistically evaluated using 2-way analysis of variance and the Duncan post hoc comparison to examine any correlation of torque generation with glide path establishment and nickel-titanium instruments. The significance level was set at 95%. RESULTS: The generated total torque by WaveOne Gold was significantly reduced by glide path establishment (P < .05), whereas glide path establishment did not induce significant changes in the maximum torque for both file systems. WaveOne Gold with a glide path showed the lowest total torque generation among all groups (P < .05). WaveOne generated a higher maximum torque than WaveOne Gold regardless of the establishment of a glide path (P < .05). CONCLUSIONS: Under the limitations of this study, glide path establishment and the mechanical property of instruments have a significant influence on torque generation. It is recommended to create the glide path and use a flexible file to reduce torque generation and, consequently, the risk of file fracture and root dentin damage.

Biofilms in Endodontics-Current Status and Future Directions.

Microbiota are found in highly organized and complex entities, known as biofilms, the characteristics of which are fundamentally different from microbes in planktonic suspensions. Root canal infections are biofilm mediated. The complexity and variability of the root canal system, together with the multi-species nature of biofilms, make disinfection of this system extremely challenging. Microbial persistence appears to be the most important factor for failure of root canal treatment and this could further have an impact on pain and quality of life. Biofilm removal is accomplished by a chemo-mechanical process, using specific instruments and disinfecting chemicals in the form of irrigants and/or intracanal medicaments. Endodontic research has focused on the characterization of root canal biofilms and the clinical methods to disrupt the biofilms in addition to achieving microbial killing. In this narrative review, we discuss the role of microbial biofilms in endodontics and review the literature on the role of root canal disinfectants and disinfectant-activating methods on biofilm removal.

Preference of undergraduate students after first experience on nickel-titanium endodontic instruments.

OBJECTIVES: This study aimed to compare two nickel-titanium systems (rotary vs. reciprocating) for their acceptance by undergraduate students who experienced nickel-titanium (NiTi) instruments for the first time. MATERIALS AND METHODS: Eighty-one sophomore dental students were first taught on manual root canal preparation with stainless-steel files. After that, they were instructed on the use of ProTaper Universal system (PTU, Dentsply Maillefer), then the WaveOne (WO, Dentsply Maillefer). They practiced with each system on 2 extracted molars, before using those files to shape the buccal or mesial canals of additional first molars. A questionnaire was completed after using each file system, seeking students' perception about 'Ease of use', 'Flexibility', 'Cutting-efficiency', 'Screwing-effect', 'Feeling-safety', and 'Instrumentation-time' of the NiTi files, relative to stainless-steel instrumentation, on a 5-point Likert-type scale. They were also requested to indicate their preference between the two systems. Data was compared between groups using t-test, and with Chi-square test for correlation of each perception value with the preferred choice (p = 0.05). RESULTS: Among the 81 students, 55 indicated their preferred file system as WO and 22 as PTU. All scores were greater than 4 (better) for both systems, compared with stainless-steel files, except for 'Screwing-effect' for PTU. The scores for WO in the categories of 'Flexibility', 'Screwing-effect', and 'Feeling-safety' were significantly higher scores than those of PTU. A significant association between the 'Screwing-effect' and students' preference for WO was observed. CONCLUSIONS: Novice operators preferred nickel-titanium instruments to stainless-steel, and majority of them opted for reciprocating file instead of continuous rotating system.

A micro-computed tomography study of the negotiation and anatomical feature in apical root canal of mandibular molars.

The aim of this study was to investigate the clinical negotiation of various apical anatomic features of the mandibular first molars in a Chinese population using micro-computed tomography (micro-CT). A total of 152 mandibular first molars were scanned with micro-CT at 30 µm resolution. The apical 5 mm of root canal (ARC) was reconstructed three dimensionally and classified. Subsequently, the access cavity was prepared with the ARC anatomy blinded to the operator. The ARC was negotiated with a size 10 K file with or without precurve. Information on the ability to obtain a reproducible glide path was recorded. The anatomical classification of ARC was Type I with 68.45% in mandibular first molars. The negotiation result of ARC with Category i was 387 canals (74.00%). With a bent negotiating file, 96 canals were negotiated, including 88 reproducible glide paths (Category ii) and 8 irregular glide paths (Category iii). About 7.65% canals could not be negotiated with patency successfully (Category iv). The statistical analyze shown the anatomic feature of ARC had effect on the negotiation of ARC (p < 0.05). In conclusion, ARC anatomic variations had a strong potential impact on the negotiation. The category of negotiation in ARC would be helpful in the using of NiTi rotary instruments. Negotiation of ARC to the working length with patency should be careful and skillful because of the complexities of ARC. SCANNING 38:819-824, 2016. © 2016 Wiley Periodicals, Inc.

Effects of Recombinant Overexpression of Bcl2 on the Proliferation, Apoptosis, and Osteogenic/Odontogenic Differentiation Potential of Dental Pulp Stem Cells.

INTRODUCTION: The therapeutic usefulness of dental pulp stem cells (DPSCs) is severely limited by low survivability upon transplantation in situ because of the presence of various proapoptotic factors within damaged/diseased tissues (ie, hypoxia and inflammation). One strategy to enhance the survivability of grafted DPSCs could be recombinant overexpression of antiapoptotic genes, such as the B-cell lymphoma 2 gene (Bcl2). METHODS: DPSCs were transfected with the Bcl2 and/or GFP gene. Cell density and mitotic activity of transfected DPSCs within in vitro culture were evaluated with the water soluble tetrazolium salt-8 (WST-8) and bromodeoxyuridine assay, respectively, whereas apoptosis was evaluated through the detection of cytoplasmic histone-associated DNA fragments. The osteogenic/odontogenic differentiation potential of these cells was evaluated with quantitative real-time polymerase chain reaction, alkaline phosphatase, and alizarin red staining. RESULTS: Bcl2-transfected DPSCs exhibited consistently higher cell densities than the GFP-transfected control within in vitro culture, and this was not because of the higher mitotic rate but was instead attributed to enhanced cell survivability because of the inhibition of apoptosis by Bcl2. Recombinant overexpression of Bcl2 inhibited the osteogenic/odontogenic potential of DPSCs, as indicated by lower levels of alkaline phosphatase activity and mineralized calcium deposition, together with the down-regulated expression of several key osteogenic/odontogenic gene markers including collagen I, osteocalcin, dentin matrix protein-1, bone sialoprotein, and alkaline phosphatase. CONCLUSIONS: The results place a "caveat" or limitation on the use of recombinant Bcl2 overexpression as a therapeutic strategy for improving the survivability of grafted DPSCs in that the osteogenic/odontogenic potential of these cells may be compromised despite enhanced survival within the host.

Quality of root canal fillings using three gutta-percha obturation techniques.

OBJECTIVES: The goal of this study was to compare the density of gutta-percha root fillings obturated with the following techniques: cold lateral (CL) compaction, ultrasonic lateral (UL) compaction, and warm vertical (WV) compaction. MATERIALS AND METHODS: Thirty-three extracted mandibular first molars, with two separate mesial canals in each, were selected. After instrumentation, the canals were stratified into three groups based on canal length and curvature, and underwent obturation with one of the techniques. No sealer was used in order to avoid masking any voids. The teeth were imaged pre- and post-obturation using micro-computed tomography. The reconstructed three-dimensional images were analyzed volumetrically to determine the amount of gutta-percha present in every 2 mm segment of the canal. P values < 0.05 were considered to indicate statistical significance. RESULTS: The overall mean volume fraction of gutta-percha was 68.51 ± 6.75% for CL, 86.56 ± 5.00% for UL, and 88.91 ± 5.16% for WV. Significant differences were found between CL and UL and between CL and WV (p < 0.05), but not between UL and WV (p = 0.526). The gutta-percha density of the roots treated with WV and UL increased towards the coronal aspect, but this trend was not noted in the CL group. CONCLUSIONS: WV compaction and UL compaction produced a significantly denser gutta-percha root filling than CL compaction. The density of gutta-percha was observed to increase towards the coronal aspect when the former two techniques were used.

Lipoteichoic acid from an Enterococcus faecalis clinical strain promotes TNF-α expression through the NF-κB and p38 MAPK signaling pathways in differentiated THP-1 macrophages.

To study the immune-inflammatory response and signaling mechanism of macrophages to purified Enterococcus faecalis (E. faecalis) lipoteichoic acid (LTA), intact LTA was obtained from an E. faecalis clinical strain P25RC using the butanol method and hydrophobic interaction chromatography purification. The fractions containing LTA were determined using phosphate detection. Contaminations with lipopolysaccharide and proteins were excluded using the Limulus amoebocyte lysate assay and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, respectively. LTA was analyzed using nuclear magnetic resonance. Prior to LTA stimulation assays, THP-1 monocytes were pretreated with phorbol 12-myristate 13-acetate to differentiate into macrophages. Macrophages were treated with LTA in concentration gradients and cells without LTA treatment as the control. Gene expression of TLR2, CD14 and MyD88 were evaluated by quantitative polymerase chain reaction. Tumor necrosis factor-α (TNF-α) and interleukin (IL)-10 were quantified using ELISA. The activated and total nuclear factor-κB (NF-κB) p65 and three mitogen-activated protein kinases (p38, ERK1/2 and JNK) were assessed using western blot analysis. E. faecalis LTA induced the gene expression of TLR2 and MyD88 whilst it downregulated CD14, suggesting a TLR2-dependent and CD14-independent immune-inflammatory activity. LTA stimulated the expression of pro-inflammatory cytokine TNF-α (P<0.05), but not the anti-inflammatory cytokine IL-10. In conclusion, E. faecalis LTA stimulated the expression of TNF-α in macrophages possibly through the NF-κB and p38 pathways.