Effect of foraminal enlargement on microcrack formation and apical transportation: a nano-CT assessment.

The aim of this study was to evaluate the foraminal enlargement and its influence on microcrack formation and apical transportation in root canals with apical curvature. Eighteen maxillary lateral incisors with apical curvature were selected by using micro-CT images. Root canals were randomly divided in two groups (n = 9) according to root canal preparation using two working lengths: 1 mm short of the apical foramen (control group) and 1 mm beyond the apical foramen (foraminal enlargement). For both groups Reciproc Blue R40 was used for root canal instrumentation. Specimens were scanned by nano-CT (UniTOM HR) before and after root canal preparation. Percentage, length, and width of microcracks, and apical transportation were assessed. Kappa, chi-square and McNemar tests were used for qualitative analyses while paired and unpaired t-test were used for quantitative analyses (α = 0.05). For both groups, rather similar and low percentages of microcracks were observed before root canal preparation (P > 0.05). The foraminal enlargement promoted new microcracks, not observed in the control group. An increase in microcrack length was observed when the foraminal enlargement was performed (P < 0.05). Higher apical transportation was observed when foraminal enlargement was performed (P < 0.05). Foraminal enlargement using a heat-treated reciprocating file size 40 promoted microcracks and higher apical transportation than root canal preparation up to 1 mm short of apical foramen.

Comparison of human dental tissue RNA extraction methods for RNA sequencing.

OBJECTIVE: The purpose of this study was to identify an efficient RNA extraction method for periodontal ligament (PDL) and dental pulp (DP) tissues to be used in RNA sequencing studies, given the increased use of these techniques in dental research and the lack of standard protocols. DESIGN: PDL and DP were harvested from extracted third molars. Total RNA was extracted with four RNA extraction kits. RNA concentration, purity and integrity were assessed by means of NanoDrop and Bioanalyzer and statistically compared. RESULTS: RNA from PDL was more likely to be degraded than that of DP. The TRIzol method yielded the highest RNA concentration from both tissues. All methods harvested RNA with A260/A280 close to 2.0 and with A260/A230 above 1.5, except for the A260/A230 from PDL obtained with the RNeasy Mini kit. For RNA integrity, the RNeasy Fibrous Tissue Mini kit yielded the highest RIN values and 28 S/18 S from PDL, while the RNeasy Mini kit obtained relatively high RIN values with an appropriate 28 S/18 S for DP. CONCLUSION: Significantly different results were obtained for PDL and DP when using the RNeasy Mini kit. The RNeasy Mini kit provided the highest RNA yields and quality for DP, while the RNeasy Fibrous Tissue Mini kit obtained the highest quality RNA from PDL.

Dynamic navigation: a laboratory study on the accuracy and potential use of guided root canal treatment.

AIM: To evaluate 3D accuracy and outcome of a dynamic navigation method for guided root canal treatment of severe pulp canal obliteration (PCO) in 3D printed jaws. METHODOLOGY: Three operators with different levels of experience in Endodontics performed navigated access cavities, using the Navident system (ClaroNav), in two sets (maxillary and mandibular) of 3D-printed jaw models with teeth presenting severe PCO. Models were mounted on a phantom to mimic a real clinical situation. After treatment, a postoperative high resolution Cone-Beam Computer Tomography (CBCT) scan (NewTom) was taken for each model and registered to the preoperative model. All access cavities were then segmented using 3-Matic Medical software 15.0 (Materialise). Length and volume of each access cavity were measured, and a comparison was done by measuring the distance deviation in mm at the coronal entry point, apical point, vertical deviation, total deviation and angular deviation of the access cavity in comparison with the virtual planning. Additionally, all access cavities were scouted with a size 10 K-file and inspected on the CBCT to confirm that the canal was located. Descriptive statistics for each parameter were performed. Normality of the data was assessed; data were transformed if needed to make it normally distributed. One-way analysis of variance (anova) was applied to assess differences between parameters for tooth type, jaw, and operators and corrected for simultaneous hypothesis testing according to Tukey. Significance level was set at .05. RESULTS: After training with the system (28 cavities per operator), a total of 132 teeth and 168 access cavities (56 per operator) were prepared. All operators located a total of 156 canals, obtaining an overall success of 93% without a difference between operator experience (p > .05). The mean deviation at the apical point was 0.63 mm (SD 0.35) and was significantly lower in anterior teeth in comparison with molars (p < .05). The mean angular deviation from the planning was 2.81° (SD 1.53). CONCLUSION: Dynamic navigation was an accurate approach for root canal treatment in teeth with severely calcified canals. However, the technique has a learning curve and requires extensive training prior to its use clinically.

Mesiobuccal Root Canal Morphology of Maxillary First Molars in a Brazilian Sub-Population - A Micro-CT Study.

Objective: This study aimed to investigate the root canal system morphology of maxillary first molar mesiobuccal (MB) roots in a Brazilian sub-population using micro-computed tomography. Methods: Ninety-six MB roots were scanned with a micro-CT (Skyscan 1173, Bruker). Three-dimensional images were analyzed regarding the number of pulp chamber orifices, the number and classification of the canals, the presence of accessory canals in different thirds of the root as well as the number and type of apical foramina. Results: A single entrance orifice was found in 53.0% of the samples, two in 43.9% and only 3.1% had three orifices. The second mesiobuccal root canal (MB2) was present at some portion of the root in 87.5% of the specimens. A single apical foramen was present in 16.7%, two in 22.9%, and three or more foramina in 60.4% of the roots. Only 55.3% and 76.1% of the root canals could be arranged by Weine's and Vertucci's classifications, respectively. Conclusion: The number of orifices at the pulp chamber level could not work as a predictor of the MB2 presence. The most prevalent canal configuration was Weine type IV / Vertucci type V. The anatomical complexity of the MB root could not be entirely classified by the current most accepted classifications.

Preclinical effectiveness of an experimental tricalcium silicate cement on pulpal repair.

OBJECTIVES: To investigate the pulpal repair potential of an experimental zirconium-oxide containing tricalcium-silicate cement, referred to as 'TCS 50'. MATERIALS AND METHODS: The effect of TCS 50 on viability, proliferation, migration, and odontoblastic differentiation of human dental pulp cells (HDPCs) was assessed using XTT assay, in-vitro wound healing assay and RT-PCR, respectively. Additionally, the pulp-capping potential was evaluated using a vital human tooth model. Statistical analysis was performed using non-parametric Kruskal-Wallis test and post-hoc test (Mann-Whitney U test). The tests were performed at a significance level of α = 0.05. RESULTS: The effect of TCS 50 towards HDPCs was dose dependent. Undiluted TCS 50 extract showed no immediate adverse impact on cell viability (p > .05); however, it significantly inhibited proliferation and migration of HDPCs (p < .05). A 25% diluted TCS 50 extract showed no significant effect on cell viability, proliferation or migration (p > .05), and it significantly enhanced odontoblastic differentiation of HDPCs (p < .05). In pulps capped with TCS 50 for both 2 and 4 weeks, H&E staining revealed a normal morphology of pulp tissue; mineralized foci with cellular components entrapped in the matrix were formed underneath the exposure site. Collagen I expression was weak within the matrix of mineralized foci, while the expression of nestin was positive for entrapped cellular components within the mineralized foci, indicating that the formed mineralized foci corresponded to an initial form of reparative dentin formation. CONCLUSION: TCS 50 is capable of generating an early pulp-healing reaction and therefore could serve as a promising pulp-capping agent.

Development of self-adhesive pulp-capping agents containing a novel hydrophilic and highly polymerizable acrylamide monomer.

Several studies have shown the clinical success of hydraulic calcium-silicate cements (hCSCs) for direct and indirect pulp capping and root repair. However, hCSCs have various drawbacks, including long setting time, poor mechanical properties, low bond strength to dentin, and relatively poor handling characteristics. To overcome these limitations, a light-curable, resin-based hCSC (Theracal LC, Bisco) was commercially introduced; however, it did not exhibit much improvement in bond strength. We developed a light-curable self-adhesive pulp-capping material that contains the novel acrylamide monomer N,N'-{[(2-acrylamido-2-[(3-acrylamidopropoxy)methyl]propane-1,3-diyl)bis(oxy)]bis(propane-1,3-diyl)}diacrylamide (FAM-401) and the functional monomer 4-methacryloxyethyl trimellitate anhydride (4-MET). Two experimental resin-based hCSCs containing different calcium sources (portlandite: Exp_Pl; tricalcium silicate cement: Exp_TCS) were prepared, and the commercial hCSCs Theracal LC and resin-free hCSC Biodentine served as controls. The performance of each cement was evaluated based on parameters relevant for vital pulp therapy, such as curing degree on a wet surface, mechanical strength, as determined using a three-point bending test, shear bond strength to dentin, cytotoxicity, as determined using an MTT assay, and the amount of calcium released, as determined using inductively coupled plasma atomic emission spectrometry. Both experimental cements cured on wet surfaces and showed relatively low cytotoxicity. Furthermore, their flexural and shear bond strength to dentin were significantly higher than those of the commercial references. High calcium release was observed for both Exp_Pl and Biodentine. Thus, Exp_Pl as a new self-adhesive pulp-capping agent performed better than the commercial resin-based pulp-capping agent in terms of mechanical strength, bond strength, and calcium release.

Injectable phosphopullulan-functionalized calcium-silicate cement for pulp-tissue engineering: An in-vivo and ex-vivo study.

OBJECTIVE: To evaluate, by means of an ex-vivo human tooth-culture model and in-vivo minipig animal study, the pulpal inflammatory reaction and reparative dentin-formation capacity of an injectable phosphopullulan-based calcium-silicate cement (GC, Tokyo, Japan) upon pulp capping, this in comparison with the commercial reference material Biodentine (Septodont). METHODS: For the ex-vivo tooth model, 9 freshly-extracted teeth from 3 different patients were pulp-capped with the experimental biomaterial (n = 3), Biodentine (n = 3) or left uncapped (control; n = 3). The teeth were kept in fresh culture medium for 4 weeks and, upon fixation three-dimensional Micro-CT and histology were performed. For the in-vivo animal study, 40 teeth from 3 minipigs were exposed and pulp capped with the experimental biomaterial containing phosphopullulan (n = 24) or Biodentine (n = 16) for 7 or 70 days. The inflammatory reaction and the tissue-regenerative potential was qualitatively and semi-quantitatively characterized using three-dimensional micro-CT and histology. RESULTS: Ex vivo, the treatment with the experimental phosphopullulan-based calcium-silicate cement and Biodentine stimulated the formation of fibrous tissue and mineralized foci. In vivo, early inflammatory reaction and regeneration of the pulp-tissue interface was promoted by both bioceramic materials after 7 and 70 days, respectively. SIGNIFICANCE: Our findings bring new insights into calcium-silicate-mediated dental pulp repair and regeneration. The novel ready-to-use and self-adhering functionalized calcium-silicate cement revealed effective pulpal repair potential.

Experimental tricalcium silicate cement induces reparative dentinogenesis.

OBJECTIVES: To overcome shortcomings of hydraulic calcium-silicate cements (hCSCs), an experimental tricalcium silicate (TCS) cement, named 'TCS 50', was developed. In vitro research showed that TCS 50 played no negative effect on the viability and proliferation of human dental pulp cells, and it induced cell odontogenic differentiation. The objective was to evaluate the pulpal repair potential of TCS 50 applied onto exposed minipig pulps. METHODS: Twenty permanent teeth from three minipigs were mechanically exposed and capped using TCS 50; half of the teeth were scheduled for 7-day and the other half for 70-day examination (n=10). Commercial hCSCs ProRoot MTA and TheraCal LC were tested as references (n=8). Tooth discoloration was examined visually. After animal sacrifice, the teeth were scanned using micro-computed tomography; inflammatory response at day 7 and day 70, mineralized tissue formation at day 70 were assessed histologically. RESULTS: Up to 70 days, TCS 50 induced no discoloration, ProRoot MTA generated gray/black discoloration in all teeth. For TCS 50, 40.0% pulps exhibited a mild/moderate inflammation at day 7. No inflammation was detected and complete reparative dentin with tubular structures was formed in all pulps after 70 days. ProRoot MTA induced a similar response, TheraCal LC generated a less favorable response in terms of initial inflammation and reparative dentin formation; however, these differences were not significant (Chi-square test of independence: p>0.05). SIGNIFICANCE: TCS 50 induced reparative dentinogenesis in minipig pulps. It can be considered as a promising pulp-capping agent, also for aesthetic areas.