Simulated Root Canal Model Research Articles

Injectable Double-Network Hydrogel-Based Three-Dimensional Cell Culture Systems for Regenerating Dental Pulp

The regeneration of dental pulp tissue is very important, but difficult, in dentistry. The biocompatibility, water content, and viscoelastic properties of pulp-like tissue must be optimized to achieve the efficient transfer of metabolites and nutrients, a suitable degradation rate, distribution of encapsulated cells, injectability, and gelation in situ under physiological conditions. As promising materials for pulp regeneration, hydrogel scaffolds have been produced to simulate the extracellular matrix and transmit signaling molecules. It is imperative to develop hydrogels to effectively regenerate pulp tissue for clinical application. Here, two injectable double-network (DN) hydrogel-based three-dimensional (3D) cell culture systems were developed for regenerating dental pulp. The microstructure, mechanical property, rheology property, and degradation behavior of the injectable DN glycol chitosan-based hydrogels in a simulated root canal model were characterized and compared to a single-network (SN) glycol chitosan-based hydrogel. Human dental pulp stem cells (hDPSCs) were then encapsulated into the GC-based hydrogels for the regeneration of pulp tissue, and the biological performance was investigated both in vitro and in vivo. The results showed that the DN hydrogels had ideal injectability under physiological conditions due to the dynamic nature of the crosslinks. Besides, the DN hydrogels exhibited better mechanical properties and longer degradation duration than the corresponding SN hydrogel. As a 3D cell culture system, the characteristics of the DN hydrogel facilitated odontogenic differentiation and mineralization of hDPSCs in vitro. Further in vivo analysis confirmed that the chemical composition, matrix stiffness, and degradation rate of the DN hydrogel matched those of pulp-like fibrous connective tissue, which might be related to Smad3 activation. These findings demonstrate that DN glycol chitosan-based hydrogels are suitable for the regeneration of pulp tissue.

Chemical and mechanical influence of root canal irrigation on biofilm removal from lateral morphological features of simulated root canals, dentine discs and dentinal tubules.

AimTo investigate the anti‐biofilm efficacy of irrigation using a simulated root canal model, the chemical effect of irrigants against biofilms grown on dentine discs and their impact on biofilm viscoelasticity, the efficacy of the irrigants in decontaminating infected dentinal tubules and the capacity of bacteria to regrow.MethodologyBiofilm removal, viscoelastic analysis of remaining biofilms and bacterial viability were evaluated using a simulated root canal model with lateral morphological features, dentine discs and a dentinal tubule model, respectively. Experiments were conducted using a two‐phase irrigation protocol. Phase 1: a modified salt solution (RISA) and sodium hypochlorite (NaOCl) were used at a low flow rate to evaluate the chemical action of the irrigants. Ultrasonic activation (US) of a chemically inert solution (buffer) was used to evaluate the mechanical efficacy of irrigation. Phase 2: a final irrigation with buffer at a high flow rate was performed for all groups. Optical coherence tomography (OCT), low load compression testing (LLCT) and confocal scanning laser microscopy analysis were used in the different models. One‐way analysis of variance (anova) was performed for the OCT and LLCT analysis, whilst Kruskal–Wallis and Wilcoxon ranked tests for the dentinal tubule model.ResultsUS and high flow rate removed significantly more biofilm from the artificial lateral canal. For biofilm removal from the artificial isthmus, no significant differences were found between the groups. Within‐group analysis revealed significant differences between the steps of the experiment, with the exception of NaOCl. For the dentine discs, no significant differences regarding biofilm removal and viscoelasticity were detected. In the dentinal tubule model, NaOCl exhibited the greatest anti‐biofilm efficacy.ConclusionsThe mechanical effect of irrigation is important for biofilm removal. An extra high flow irrigation rate resulted in greater biofilm removal than US in the artificial isthmus. The mechanical effect of US seemed to be more effective when the surface contact biofilm–irrigant was small. After the irrigation procedures, the remaining biofilm could survive after a 5‐day period. RISA and NaOCl seemed to alter post‐treatment remaining biofilms.

Effect of tip insertion depth and irradiation parameters on the efficacy of cleaning calcium hydroxide from simulated lateral canals using Er:YAG laser- or ultrasonic-activated irrigation

Background/purposeLaser-activated root canal irrigation (LAI) with an Er:YAG laser is considered more effective than other irrigation methods, whereas the effectiveness of LAI in cleaning lateral canals far from the laser tip remains unclear. This study aimed to compare the efficacy of removing calcium hydroxide [Ca(OH)2] paste from lateral canals using LAI or ultrasonic-activated irrigation (UAI), and to examine the effect of tip insertion depth and laser irradiation parameters on cleaning efficacy. Materials and methodsRadiopaque Ca(OH)2 paste (Calcipex II) was injected into lateral canals 6 mm from the root apex in 192 J-shaped simulated root canal models. LAI (Erwin AdvErl; 30 or 70 mJ; 10 or 20 pulses per second; laser tip R200T or R600T) and UAI (ENAC SE10; output setting: 3) were performed 3 times for 20 s. The laser tip was placed at 8–0 mm coronal to the lateral canal location. The volume of Ca(OH)2 paste before and after the experiment was measured using micro-CT (SMX-100CT). ResultsThe Ca(OH)2 removal rate by LAI was significantly higher than UAI at all tip insertion depths. Ca(OH)2 removal rate in LAI was significantly lower at the 8 mm position compared with other positions (P < 0.05). When the tip insertion depth was fixed at this position, Ca(OH)2 removal rate increased significantly when pulse energy and tip diameter were increased (P < 0.05). ConclusionLAI removed Ca(OH)2 paste from lateral canals away from the tip more effectively than UAI. Increasing the pulse energy and tip diameter improved the removal efficiency.

Novel Activated Microbubbles-based Strategy to Coat Nanoparticles on Root Canal Dentin: Fluid Dynamical Characterization

IntroductionActivated microbubbles (MBs) have the potential to deliver nanoparticles in complex microspaces such as root canals. The objective of the study is to determine the fluid dynamical parameters associated with ultrasonic, sonic, and manual activation of MBs in simulated root canals and to assess the effectiveness of surface coating formed by delivering chitosan nanoparticles using activated MBs within root canals in extracted teeth. MethodsIn stage 1, polydimethylsiloxane models were fabricated to determine the physical effects of MBs agitated manually (MM), sonically (MS), and ultrasonically (MU). Spherical tracer particles were used to visualize and record the fluid motion using an inverted microscope linked to a high-speed camera. The velocity, wall stress, and penetration depth were analyzed at regions of interest. In stage 2, 35 extracted human incisors were divided into 7 groups to evaluate the effectiveness of chitosan nanoparticle delivery using activated MBs (MM, MS, and MU groups). Field emission scanning electron microscopy and energy-dispersive X-rays were used to characterize the nanoparticle coating on root canal dentin and the degree of dentinal tubule occlusion. ResultsIn stage 1, velocity, wall stress, and penetration depth increased significantly in the MB groups compared with the control (P < .01). In stage 2, 70% of the dentin surface was coated, and 65% of the dentinal tubule was occluded with nanoparticle-based coating in the MM, MU, and water ultrasonic groups. Element analysis displayed the presence of dentin smear on the root canal surface for the MU and water ultrasonic groups. ConclusionsActivated MBs enhanced fluid dynamical parameters when compared with water in simulated root canal models. Manual activation of MBs resulted in uniform and significant nanoparticle-based surface coating and tubule blockage in root canal dentin without dentin smear formation.

Investigations into insitu Enterococcus faecalis biofilm removal by passive and active sodium hypochlorite irrigation delivered into the lateral canal of a simulated root canal model.

To investigate insitu Enterococcus faecalis biofilm removal from the lateral canal of a simulated root canal system using passive or active irrigation protocols. Root canal models (n=43) were manufactured from transparent resin materials using 3D printing. Each canal was created with an 18mm length, apical size 30, a .06 taper and a lateral canal of 3mm length, with 0.3mm diameter. Biofilms were grown in the lateral canal and apical 3mm of the main canal for 10days. Three models from each group were examined for residual biofilm using SEM. The other forty models were divided into four groups (n=10). The models were observed under a fluorescence microscope. Following 60s of 9mL of 2.5% NaOCl irrigation using syringe and needle, the irrigant was either left stagnant in the canal or activated using gutta-percha, sonic or ultrasonic methods for 30s. Images were then captured every second using an external camera. The residual biofilm percentages were measured using image analysis software. The data were analysed using generalized linear mixed models. A significance level of 0.05 was used throughout. The greatest level of biofilm removal was obtained with ultrasonic agitation (66.76%) followed by sonic (45.49%), manual agitation (43.97%) and passive irrigation groups (38.67%), respectively. The differences were significant between the residual biofilm in the passive irrigation and both sonic and ultrasonic groups (P=0.001). Agitation resulted in better penetration of 2.5% NaOCl into the lateral canal of an artificial root canal model. Ultrasonic agitation of NaOCl improved the removal of biofilm.

The effect of sodium hypochlorite concentration and irrigation needle extension on biofilm removal from a simulated root canal model.

To investigate the effect of sodium hypochlorite concentration and needle extension on removal of Enterococcus faecalis biofilm, sixty root canal models were 3D printed. Biofilms were grown on the apical 3 mm of the canal for 10 days. Irrigation for 60s with 9 mL of either 5.25% or 2.5% NaOCl or water was performed using a needle inserted either 3 or 2 mm from the canal terminus and imaged using fluorescence microscopy and residual biofilm percentages were calculated using imaging software. The data were analysed using analysis of covariance and two-sample t-tests. A significance level of 0.05 was used throughout. Residual biofilm was less using 5.25% than with 2.5% NaOCl. Statistically significant biofilm removal was evident with the needle placed closer to the canal terminus. A greater reduction of available chlorine and pH was noted as the concentration increased. One-minute irrigation was not sufficient for complete biofilm removal.

Confocal laser scanning, scanning electron, and transmission electron microscopy investigation of Enterococcus faecalis biofilm degradation using passive and active sodium hypochlorite irrigation within a simulated root canal model.

Root canal irrigation is an important adjunct to control microbial infection. The aim of this study was to investigate the effect of 2.5% (wt/vol) sodium hypochlorite (NaOCl) agitation on the removal, killing, and degradation of Enterococcus faecalis biofilm. A total of 45 root canal models were manufactured using 3D printing with each model comprising an 18 mm length simulated root canal of apical size 30 and taper 0.06. E. faecalis biofilms were grown on the apical 3 mm of the models for 10 days. A total of 60 s of 9 ml of 2.5% NaOCl irrigation using syringe and needle was performed, the irrigant was either left stagnant in the canal or agitated using manual (Gutta‐percha), sonic, and ultrasonic methods for 30 s. Following irrigation, the residual biofilms were observed using confocal laser scanning, scanning electron, and transmission electron microscopy. The data were analyzed using one‐way ANOVA with Dunnett post hoc tests at a level of significance p ≤ .05. Consequence of root canal irrigation indicate that the reduction in the amount of biofilm achieved with the active irrigation groups (manual, sonic, and ultrasonic) was significantly greater when compared with the passive and untreated groups (p < .05). Collectively, finding indicate that passive irrigation exhibited more residual biofilm on the model surface than irrigant agitated by manual or automated (sonic, ultrasonic) methods. Total biofilm degradation and nonviable cells were associated with the ultrasonic group.

A novel experimental approach to investigate the effect of different agitation methods using sodium hypochlorite as an irrigant on the rate of bacterial biofilm removal from the wall of a simulated root canal model

ObjectiveRoot canal irrigation is an important adjunct to control microbial infection. This study aimed primarily to develop a transparent root canal model to study in situ Enterococcus faecalis biofilm removal rate and remaining attached biofilm using passive or active irrigation solution for 90s. The change in available chlorine and pH of the outflow irrigant were assessed. MethodsA total of forty root canal models (n=10 per group) were manufactured using 3D printing. Each model consisted of two longitudinal halves of an 18mm length simulated root canal with size 30 and taper 0.06. E. faecalis biofilms were grown on the apical 3mm of the models for 10days in Brain Heart Infusion broth. Biofilms were stained using crystal violet for visualization. The model halves were reassembled, attached to an apparatus and observed under a fluorescence microscope. Following 60s of 9mL of 2.5% NaOCl irrigation using syringe and needle, the irrigant was either left stagnant in the canal or activated using gutta-percha, sonic and ultrasonic methods for 30s. Images were then captured every second using an external camera. The residual biofilm percentages were measured using image analysis software. The data were analyzed using Kruskal–Wallis test and generalized linear mixed model. ResultsThe highest level of biofilm removal was with ultrasonic agitation (90.13%) followed by sonic (88.72%), gutta-percha (80.59%), and passive irrigation group (control) (43.67%) respectively. All agitation groups reduced the available chlorine and pH of NaOCl more than that in the passive irrigation group. SignificanceThe 3D printing method provided a novel model to create a root canal simulation for studying and understanding a real-time biofilm removal under microscopy. Ultrasonic agitation of NaOCl left the least amount of residual biofilm in comparison to sonic and gutta-percha agitation methods.

An in vitro Evaluation on a Novel Root Canal Cleansing Method by Using Nylon Fibers

Despite traditional metal-based dental files, such as NiTi being demonstrated effective in root cleaning, the tooth structure is always damaged. Thus, to fulfill the need for a minimally invasive tool for contemporary endodontics and dentistry, the use of polymer fibers might provide a good option, as it is soft, fabricable, and disposable. In this study, two types of nylon fibers with respective average diameters of 206.9 µm (fiber W) and 156.4 µm (fiber B), respectively, were used as dental files, and mounted onto either a reciprocating or a low-speed rotary hand-piece. In vitro, simulated root canal models were colored red using nail varnish, and then cleaned by the fiber files mounted on the hand-pieces. Three parts of the simulated models, i.e., the apical third, the medium third, and the coronal third, were chosen to assess the cleaning the efficiency (CE) of each specimen by calculating the ratio of the cross-sectional area changes, before and after cleansing, using micro-Computer Tomography (CT). A NiTi file with a low-speed hand-piece was used as a control. SEM was used to observe the nylon fiber surfaces before and after the cleansing. Micro-CT results showed that for both the nylon fibers, W and B, an average CE of 82.11% ± 9.68% for the medium third could be achieved, which is statistically higher (p &lt; 0.01) than the coronal third and apical third. The cleansing efficiency was not affected by, the types of fibers, nor the hand-pieces according to student’s t-test. Most of the nylon fibers could withstand deformation after the cleansing. To conclude, nylon fiber files have demonstrated a certain cleansing efficiency in simulated root canals, and micro-CT is a promising method to assess CE.

Fluid Dynamics and Biofilm Removal Generated by Syringe-delivered and 2 Ultrasonic-assisted Irrigation Methods: A Novel Experimental Approach

IntroductionThorough understanding of fluid dynamics in root canal irrigation and corresponding antibiofilm capacity will support improved disinfection strategies. This study aimed to develop a standardized, simulated root canal model that allows real-time analysis of fluid/irrigation dynamics and its correlation with biofilm elimination. MethodsA maxillary incisor with an instrumented root canal was imaged with micro–computed tomography. The canal volume was reconstructed in 3 dimensions and replicated in soft lithography-based models microfabricated from polyethylene glycol–modified polydimethylsiloxane. Canals were irrigated by using a syringe (SI) and 2 ultrasonic-assisted methods, intermittent (IUAI) and continuous (CUAI). Real-time fluid movement within the apical 3 mm of canals was imaged by using microparticle image velocimetry. In similar models, canals were inoculated with Enterococcus faecalis to grow 3-week-old biofilms. Biofilm reduction by irrigation with SI, CUAI, and IUAI was assessed by using a crystal violet assay and compared with an untreated control. ResultsSI generated higher velocity and shear stress in the apical 1–2 mm than 0–1 and 2–3 mm. IUAI generated consistently low shear stress in the apical 3 mm. CUAI generated consistently high levels of velocity and shear stress; it was the highest of the groups in the apical 0–1 and 2–3 mm. Biofilm was significantly reduced compared with the control only by CUAI (two-sample permutation test, P = .005). ConclusionsCUAI exhibited the highest mechanical effects of fluid flow in the apical 3 mm, which correlated with significant biofilm reduction. The soft lithography-based models provided a novel model/method for study of correlations between fluid dynamics and the antibiofilm capacity of root canal irrigation methods.

Measurement and visualization of file‐to‐wall contact during ultrasonically activated irrigation in simulated canals

(i) To quantify in a simulated root canal model the file-to-wall contact during ultrasonic activation of an irrigant and to evaluate the effect of root canal size, file insertion depth, ultrasonic power, root canal level and previous training, (ii) To investigate the effect of file-to-wall contact on file oscillation. File-to-wall contact was measured during ultrasonic activation of the irrigant performed by 15 trained and 15 untrained participants in two metal root canal models. Results were analyzed by two 5-way mixed-design anovas. The level of significance was set at P<0.05. Additionally, high-speed visualizations, laser-vibrometer measurements and numerical simulations of the file oscillation were conducted. File-to-wall contact occurred in all cases during 20% of the activation time. Contact time was significantly shorter at high power (P<0.001), when the file was positioned away from working length (P<0.001), in the larger root canal (P<0.001) and from coronal towards apical third of the root canal (P<0.002), in most of the cases studied. Previous training did not show a consistent significant effect. File oscillation was affected by contact during 94% of the activation time. During wall contact, the file bounced back and forth against the wall at audible frequencies (ca. 5kHz), but still performed the original 30kHz oscillations. Travelling waves were identified on the file. The file oscillation was not dampened completely due to the contact and hydrodynamic cavitation was detected. Considerable file-to-wall contact occur-red during irrigant activation. Therefore, the term 'Passive Ultrasonic Irrigation' should be amended to 'Ultrasonically Activated Irrigation'.

Elimination of E. Faecalis by a New Non‐Thermal Atmospheric Pressure Plasma Handheld Device for Endodontic Treatment. A Preliminary Investigation

AbstractThe aim of this study was to investigate the feasibility of an atmospheric pressure plasma jet (PJ) for endodontic treatment and to compare the antimicrobial efficiency with conventional irrigation solutions (chlorhexidine digluconate (CHX), sodium hypochlorite (NaOCl)) against intracanal bacteria (E. faecalis) using a standardized simulated root canal model. The disinfection efficiency was determined by number of colony forming units. Scanning electron microscopy was used to investigate the effect of the treatment procedures on bacteria inside the root canal. The PJ achieved a significantly higher microbial reduction than chemical CHX irrigation and a comparable one with NaOCl. The application of PJ for endodontic treatment could provide a new possibility of controlling endodontic infection. magnified image

An In Vitro Comparative Study of Intracanal Fluid Motion and Wall Shear Stress Induced by Ultrasonic and Polymer Rotary Finishing Files in a Simulated Root Canal Model

Objective. This in vitro study compared the flow pattern and shear stress of an irrigant induced by ultrasonic and polymer rotary finishing file activation in an acrylic root canal model. Flow visualization analysis was performed using an acrylic canal filled with a mixture of distilled water and rheoscopic fluid. The ultrasonic and polymer rotary finishing file were separately tested in the canal and activated in a static position and in a cyclical axial motion (up and down). Particle movement in the fluid was captured using a high-speed digital camera and DaVis 7.1 software. The fluid shear stress analysis was performed using hot film anemometry. A hot-wire was placed in an acrylic root canal and the canal was filled with distilled water. The ultrasonic and polymer rotary finishing files were separately tested in a static position and in a cyclical axial motion. Positive needle irrigation was also tested separately for fluid shear stress. The induced wall shear stress was measured using LabVIEW 8.0 software.

Debris retention and wear in three different nickel‐titanium rotary instruments

The aim of this in vitro study was to compare three nickel-titanium (Ni-Ti) rotary instruments: EndoWave, K3 and ProFile, with respect to their debris retention and wear. Thirty simulated root canal models made from resin blocks with 40 degrees canal curvature were used. They were divided into three groups of 10 samples each; EndoWave, K3 and ProFile were used in Groups A, B and C respectively. They were prepared with four serial size files. The weight of each file was measured to evaluate debris retention and wear at four representative time points: prior to root canal preparation (W1), immediately after root canal preparation (W2), after ultrasonic cleaning (W3), after further additional cleaning (W4). The weight of debris on each file after root canal instrumentation and ultrasonic cleaning was calculated as D1: (W2 - W4) and D2: (W3 - W4) respectively. Also, wear of the file was calculated as WF: (W1 - W4). There was a significant difference in the amount of debris remaining on the file among the three files; EndoWave retained more debris (D1) than K3 and ProFile (P < 0.001, one-way anova). ProFile had the largest amount of debris (D2) remaining after ultrasonic cleaning (P < 0.001). Wear was not detectable in this study. Under the conditions of this study, immediately after root canal preparation, EndoWave retained the largest amount of debris, while ProFile retained the most debris even after ultrasonic cleaning.