Brian E. Bergeron

Quality of obturation achieved by an endodontic core-carrier system with crosslinked gutta-percha carrier in single-rooted canals

OBJECTIVES The present study examined the quality of obturation in root canals obturated by GuttaCore, a gutta-percha-based core-carrier system with a cross-linked thermoset gutta-percha carrier, by comparing the incidence of gaps and voids identified from similar canals obturated by cold lateral compaction or warm vertical compaction. METHODS Thirty single-rooted premolars with oval-shaped canals were shaped and cleaned, and obturated with one of the three obturation techniques (N=10): GuttaCore, warm vertical compaction or cold lateral compaction. Filled canals were scanned with micro-computed tomography (micro-CT); reconstructed images were analysed for the volumetric percentage of gaps and voids at three canal levels (0-4mm, 4-8mm and 8-12mm from working length). The roots were subsequently sectioned at the 4-mm, 8-mm and 12-mm levels for analyses of the percentage of interfacial gaps, and area percentage of interfacial and intracanal voids, using scanning electron microscopy (SEM) to examine negative replicas of root sections. Data were analysed with parametric or non-parametric statistical methods at α=0.05. RESULTS Both micro-CT and SEM data indicated that canals obturated with GuttaCore core-carriers had the lowest incidence of interfacial gaps and voids, although the results were not significantly different from canals obturated by warm vertical compaction. Both the GuttaCore and the warm vertical compaction groups, in turn, had significantly lower incidences of gaps and voids than the cold lateral compaction group. CONCLUSIONS Because of the similarity in obturation quality between GuttaCore and warm vertical compaction, practitioners may find the GuttaCore core-carrier technique a valuable alternative for obturation of oval-shaped canals. CLINICAL SIGNIFICANCE The quality of obturation achieved by GuttaCore in single-rooted canals is not significantly different from that achieved by warm vertical compaction.

Efficacy of 3D conforming nickel titanium rotary instruments in eliminating canal wall bacteria from oval-shaped root canals

OBJECTIVES To evaluate the effectiveness of TRUShape® 3D Conforming Files, compared with Twisted Files, in reducing bacteria load from root canal walls, in the presence or absence of irrigant agitation. METHODS Extracted human premolars with single oval-shaped canals were infected with Enterococcus faecalis. Teeth in Group I (N=10; NaOCl and QMix® 2in1 as respective initial and final irrigants) were subdivided into 4 subgroups: (A) TRUShape® instrumentation without irrigant activation; (B) TRUShape® instrumentation with sonic irrigant agitation; (C) Twisted Files without irrigant agitation; (D) Twisted Files with sonic irrigant agitation. To remove confounding factor (antimicrobial irrigants), teeth in Group II (N=10) were irrigated with sterile saline, using the same subgroup designations. Specimens before and after chemomechanical débridement were cultured for quantification of colony-forming units (CFUs). Data from each group were analyzed separately using two-factor ANOVA and Holm-Sidak multiple comparison (α=0.05). Canal wall bacteria were qualitatively examined using scanning electron microscopy (SEM) and light microscopy of Taylor-modified Brown and Brenn-stained demineralised sections. RESULTS CFUs from subgroups in Group I were not significantly different (P=0.935). For Group II, both file type (P<0.001) and irrigant agitation (P<0.001) significantly affected log-reduction in CFU concentrations. The interaction of these two factors was not significant (P=0.601). Although SEM showed reduced canal wall bacteria, bacteria were present within dentinal tubules after rotary instrumentation, as revealed by light microscopy of longitudinal root sections. CONCLUSIONS TRUShape® files removed significantly more canal wall bacteria than Twisted Files when used without an antibacterial irrigant; the latter is required to decontaminate dentinal tubules. CLINICAL SIGNIFICANCE Root canal disinfection should not be focused only on a mechanistic approach. Rather, the rational choice of a rotary instrumentation system should be combined with the use of well-tested antimicrobial irrigants and delivery/agitation techniques to establish a clinically realistic chemomechanical débridement protocol.

Potential applications of antimicrobial peptides and their mimics in combating caries and pulpal infections

Antimicrobial peptides (AMPs) are short cationic host-defense molecules that provide the early stage of protection against invading microbes. They also have important modulatory roles and act as a bridge between innate and acquired immunity. The types and functions of oral AMPs were reviewed and experimental reports on the use of natural AMPs and their synthetic mimics in caries and pulpal infections were discussed. Natural AMPs in the oral cavity, predominantly defensins, cathelicidins and histatins, possess antimicrobial activities against oral pathogens and biofilms. Incomplete debridement of microorganisms in root canal space may precipitate an exacerbated immune response that results in periradicular bone resorption. Because of their immunomodulatory and wound healing potentials, AMPs stimulate pro-inflammatory cytokine production, recruit host defense cells and regulate immuno-inflammatory responses in the vicinity of the pulp and periapex. Recent rapid advances in the development of synthetic AMP mimics offer exciting opportunities for new therapeutic initiatives in root canal treatment and regenerative endodontics. STATEMENT OF SIGNIFICANCE Identification of new therapeutic strategies to combat antibiotic-resistant pathogens and biofilm-associated infections continues to be one of the major challenges in modern medicine. Despite the presence of commercialization hurdles and scientific challenges, interests in using antimicrobial peptides as therapeutic alternatives and adjuvants to combat pathogenic biofilms have never been foreshortened. Not only do these cationic peptides possess rapid killing ability, their multi-modal mechanisms of action render them advantageous in targeting different biofilm sub-populations. These factors, together with adjunctive bioactive functions such as immunomodulation and wound healing enhancement, render AMPs or their synthetic mimics exciting candidates to be considered as adjuncts in the treatment of caries, infected pulps and root canals.

Effects of a discoloration-resistant calcium aluminosilicate cement on the viability and proliferation of undifferentiated human dental pulp stem cells

Discoloration-resistant calcium aluminosilicate cement has been formulated to overcome the timely problem of tooth discoloration reported in the clinical application of bismuth oxide-containing hydraulic cements. The present study examined the effects of this experimental cement (Quick-Set2) on the viability and proliferation of human dental pulp stem cells (hDPSCs) by comparing the cellular responses with commercially available calcium silicate cement (white mineral trioxide aggregate; WMTA) after different aging periods. Cell viability and proliferation were examined using assays that examined plasma membrane integrity, leakage of cytosolic enzyme, caspase-3 activity for early apoptosis, oxidative stress, mitochondrial metabolic activity and intracellular DNA content. Results of the six assays indicated that both Quick-Set2 and WMTA were initially cytotoxic to hDPSCs after setting for 24 h, with Quick-Set2 being comparatively less cytotoxic than WMTA at this stage. After two aging cycles, the cytotoxicity profiles of the two hydraulic cements were not significantly different and were much less cytotoxic than the positive control (zinc oxide–eugenol cement). Based on these results, it is envisaged that any potential beneficial effect of the discoloration-resistant calcium aluminosilicate cement on osteogenesis by differentiated hDPSCs is more likely to be revealed after outward diffusion and removal of its cytotoxic components.

Effect of Repeated Simulated Clinical Use and Sterilization on the Cutting Efficiency and Flexibility of Hyflex CM Nickel-Titanium Rotary Files

INTRODUCTION Recent nickel-titanium manufacturing processes have resulted in an alloy that remains in a twinned martensitic phase at operating temperature. This alloy has been shown to have increased flexibility with added tolerance to cyclic and torsional fatigue. The aim of this study was to assess the effect of repeated simulated clinical use and sterilization on cutting efficiency and flexibility of Hyflex CM rotary files. METHODS Cutting efficiency was determined by measuring the load required to maintain a constant feed rate while instrumenting simulated canals. Flexibility was determined by using a 3-point bending test. Files were autoclaved after each use according to the manufacturers recommendations. Files were tested through 10 simulated clinical uses. For cutting efficiency, mean data were analyzed by using multiple factor analysis of variance and the Dunnett post hoc test (P < .05). For flexibility, mean data were analyzed by using Levenes Test of Equality of Error and a general linear model (P < .05). RESULTS No statistically significant decrease in cutting efficiency was noted in groups 2, 5, 6, and 7. A statistically significant decrease in cutting efficiency was noted in groups 3, 4, 8, 9, and 10. No statistically significant decrease in flexibility was noted in groups 2, 3, and 7. A statistically significant decrease in flexibility was noted in groups 4, 5, 6, 8, 9, 10, and 11. CONCLUSIONS Repeated simulated clinical use and sterilization showed no effect on cutting efficiency through 1 use and no effect on flexibility through 2 uses.

Mechanism of bioactive molecular extraction from mineralized dentin by calcium hydroxide and tricalcium silicate cement

OBJECTIVES The objective of the present study was to elucidate the mechanism of bioactive molecule extraction from mineralized dentin by calcium hydroxide (Ca(OH)2) and tricalcium silicate cements (TSC). METHODS AND RESULTS Transmission electron microscopy was used to provide evidence for collagen degradation in dentin surfaces covered with Ca(OH)2 or a set, hydrated TSC for 1-3 months. A one micron thick collagen degradation zone was observed on the dentin surface. Fourier transform-infrared spectroscopy was used to identify increases in apatite/collagen ratio in dentin exposed to Ca(OH)2. Using three-point bending, dentin exposed to Ca(OH)2 exhibited significant reduction in flexural strength. Using size exclusion chromatography, it was found that the small size of the hydroxyl ions derived from Ca(OH)2 enabled those ions to infiltrate the intrafibrillar compartment of mineralized collagen and degrade the collagen fibrils without affecting the apatite minerals. Using ELISA, TGF-β1 was found to be extracted from dentin covered with Ca(OH)2 for 3 months. Unlike acids that dissolve the mineral component of dentin to release bioactive molecules, alkaline materials such as Ca(OH)2 or TSC released growth factors such as TGF-β1 via collagen degradation. SIGNIFICANCE The bioactive molecule extraction capacities of Ca(OH)2 and TSC render these dental materials excellent for pulp capping and endodontic regeneration. These highly desirable properties, however, appear to be intertwined with the untoward effect of degradation of the collagen matrix within mineralized dentin, resulting in reduced flexural strength.

Mineralogenic characteristics of osteogenic lineage-committed human dental pulp stem cells following their exposure to a discoloration-free calcium aluminosilicate cement

OBJECTIVES An experimental discoloration-free calcium aluminosilicate cement has been developed with the intention of maximizing the beneficial attributes of tricalcium silicate cements and calcium aluminate cements. The present study examined the effects of this experimental cement (Quick-Set2) on the mineralogenic characteristics of osteogenic lineage-committed human dental pulp stem cells (hDPSCs), by comparing the cellular responses with a commercially available tricalcium silicate cement (white mineral trioxide aggregate (ProRoot(®) MTA); WMTA). METHODS The osteogenic potential of hDPSCs exposed to the cements was examined using qRT-PCR for osteogenic gene expressions, Western blot for osteogenic-related protein expressions, alkaline phosphatase enzyme activity, Alizarin red S staining, Fourier transform infrared spectroscopy and transmission electron microscopy of extracellular calcium deposits. RESULTS Results of the six assays indicated that osteogenic differentiation of hDPSCs was significantly enhanced after exposure to the tricalcium silicate cement or the experimental calcium aluminosilicate cement, with the former demonstrating better mineralogenic stimulation capacity. SIGNIFICANCE The better osteogenic stimulating effect of the tricalcium silicate cement on hDPSCs may be due to its relatively higher silicate content, or higher OH(-) and Ca(2+) release. Further investigations with the use of in vivo animal models are required to validate the potential augmenting osteogenic effects of the experimental discoloration-free calcium aluminosilicate cement.

Antimicrobial efficacy of an apical negative pressure root canal irrigation system against intracanal microorganisms

OBJECTIVES The effect of irrigation time on the antimicrobial efficacy of an apical negative pressure irrigation system was examined in vitro, followed by validation of the antimicrobial effect in vivo using the identified optimal irrigation time. METHODS For the in vitro experiment, 44 extracted premolars were decoronated, instrumented, autoclaved and inoculated with Enterococcus faecalis (ATCC 29212) for 21 days. Four teeth were used as positive control, without irrigation. Each of the remaining 40 teeth was irrigated with 2.5% NaOCl, delivered via the EndoVac MacroCannula for 10 s, and subsequently via the EndoVac MicroCannula for 15, 30, 45, 60 or 90 s per canal, respectively (N = 8). After irrigation, microbial samples were collected, transferred to BHI broth and incubated for counting of bacterial colony forming units (CFUs). Based on the in vitro results, 8.25% NaOCl was delivered via the EndoVac MicroCannula for 60 s, during root canal treatment of 20 human subjects presented with apical periodontitis. Microbial samples retrieved in vivo prior to canal instrumentation (S0), after chemomechanical debridement (S1) and after irrigation with EndoVac (S2) were cultured in an anaerobic chamber for 7 days for CFU evaluation. RESULTS Compared with the control, irrigation significantly reduced bacterial populations (p < .05). Irrigation delivery via the EndoVac demonstrated improved antibacterial efficacy with increased irrigation time (p < .05). Samples retrieved from canals after NaOCl delivery in vivo with the EndoVac for 60 s were all culture-negative. CONCLUSIONS Microbial elimination may be achieved with 8.25% NaOCl delivered via the EndoVac apical negative pressure irrigation device for 60 s. CLINICAL SIGNIFICANCE With the use of the EndoVac apical negative pressure irrigant delivery system, optimal elimination of the intracanal bacterial load can only be achieved when sodium hypochlorite is delivered via the MicroCannula for at least 60 s per canal.

Antibacterial efficacy of an endodontic sonic-powered irrigation system: An in vitro study

OBJECTIVE To evaluate the efficacy of EDDY, a new sonic-powered irrigation system, in reducing intracanal bacteria load. METHODS Thirty-eight instrumented, autoclaved single-rooted human premolars were inoculated with Enterococcus faecalis (ATCC-29212) for 21 days. Two teeth were used as negative control without bacterial contamination. For the bacteria-inoculated teeth, 6 were used as positive control without irrigation. The remaining 30 teeth were randomly divided into 2 groups (N = 15), using 3% NaOCl as irrigant: (A) 30-gauge syringe needle irrigation (SNI), (B) EDDY (VDW, Munich, Germany). Twelve teeth per group and 4 teeth in the positive control were evaluated for bacterial reduction using MTT assay. The remaining teeth were split for BacLight LIVE/DEAD staining to examine the percentages of live/dead bacteria present in the dentinal tubules from different canal locations (coronal, mid-root and apical portions of the canal space) using confocal laser scanning microscopy (CLSM). RESULTS MTT assay indicated that both SNI and EDDY significantly reduced overall intracanal bacterial load compared with the positive control, with no significant difference between the two techniques. CLSM indicated that EDDY had better intratubular bacterial killing efficacy than SNI in the coronal and mid-root portions of the canal space only but not in the apical portion. In all canal locations (coronal, mid-root apical), both systems failed to eliminate bacteria that proliferated deep within the dentinal tubules. CONCLUSION With the use of 3% NaOCl, sonic-powered irrigant activation with EDDY tips did not provide additional advantage over SNI in killing Enterococcus faecalis from deep intraradicular dentin. CLINICAL SIGNIFICANCE Both the sonic-powered root canal irrigant activation system and syringe needle irrigation can reduce intracanal bacteria load but are incapable of completely killing all bacteria that resided deep within the dentinal tubules of root canals infected with Enterococcus faecalis.

Contribution of Mitophagy to Cell-Mediated Mineralization: Revisiting a 50-Year-Old Conundrum

Abstract Biomineralization in vertebrates is initiated via amorphous calcium phosphate (ACP) precursors. These precursors infiltrate the extracellular collagen matrix where they undergo phase transformation into intrafibrillar carbonated apatite. Although it is well established that ACP precursors are released from intracellular vesicles through exocytosis, an unsolved enigma in this cell‐mediated mineralization process is how ACP precursors, initially produced in the mitochondria, are translocated to the intracellular vesicles. The present study proposes that mitophagy provides the mechanism for transfer of ACP precursors from the dysfunctioned mitochondria to autophagosomes, which, upon fusion with lysosomes, become autolysosomes where the mitochondrial ACP precursors coalesce to form larger intravesicular granules, prior to their release into the extracellular matrix. Apart from endowing the mitochondria with the function of ACP delivery through mitophagy, the present results indicate that mitophagy, triggered upon intramitochondrial ACP accumulation in osteogenic lineage‐committed mesenchymal stem cells, participates in the biomineralization process through the BMP/Smad signaling pathway.