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

AIM: To 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. METHODOLOGY: Biofilm 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. RESULTS: US 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. CONCLUSIONS: The 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.

Biofilm removal from a simulated isthmus and lateral canal during syringe irrigation at various flow rates: a combined experimental and Computational Fluid Dynamics approach.

AIM: (i) To quantify biofilm removal from a simulated isthmus and a lateral canal in an artificial root canal system during syringe irrigation with NaOCl at different concentrations and delivered at various flow rates (ii) to examine whether biofilm removal is further improved by a final high-flow-rate rinse with an inert irrigant following irrigation with NaOCl. (iii) to simulate the irrigant flow in these areas using a computer model (iv) to examine whether the irrigant velocity calculated by the computer model is correlated to biofilm removal. METHODOLOGY: Ninety-six artificial root canals with either a simulated isthmus or lateral canal were used. A dual-species in vitro biofilm was formed in these areas using a Constant Depth Film Fermenter. NaOCl at various concentrations (2, 5 and 10%) or adhesion buffer (control) was delivered for 30 s by a syringe and an open-ended needle at 0.033, 0.083, or 0.166 mL s-1 or passively deposited in the main root canal (phase 1). All specimens were subsequently rinsed for 30 s with adhesion buffer at 0.166 mL s-1 (phase 2). The biofilm was scanned by Optical Coherence Tomography to determine the percentage of the remaining biofilm. Results were analysed by two 3-way mixed-design ANOVAs (α = 0.05). A Computational Fluid Dynamics model was used to simulate the irrigant flow inside the artificial root canal system. RESULTS: The flow rate during phase 1 and additional irrigation during phase 2 had a significant effect on the percentage of the remaining biofilm in the isthmus (P = 0.004 and P < 0.001). Additional irrigation during phase 2 also affected the remaining biofilm in the lateral canal significantly (P ≤ 0.007) but only when preceded by irrigation at medium or high flow rate during phase 1. The effect of NaOCl concentration was not significant (P > 0.05). Irrigant velocity in the isthmus and lateral canal increased with increasing flow rate and it was substantially correlated to biofilm removal from those areas. CONCLUSIONS: The irrigant flow rate affected biofilm removal in vitro more than NaOCl concentration. Irrigant velocity predicted by the computer model corresponded with the pattern of biofilm removal from the simulated isthmus and lateral canal.

The influence of time and irrigant refreshment on biofilm removal from lateral morphological features of simulated root canals.

AIM: To evaluate the effect of irrigant refreshment and exposure time of a 2% sodium hypochlorite solution (NaOCl) on biofilm removal from simulated lateral root canal spaces using two different flow rates. METHODOLOGY: A dual-species biofilm was formed by a Constant Depth Film Fermenter (CDFF) for 96 h in plug inserts with anatomical features resembling an isthmus or lateral canal-like structures. The inserts were placed in a root canal model facing the main canal. NaOCl 2% and demineralized water (control group) were used as irrigant solutions. Both substances were applied at a flow rate of 0.05 and 0.1 mL s-1 . The samples were divided into three groups with zero, one or two refreshments in a total exposure time of 15 min. A three-way analysis of variance (anova) was performed to investigate the interaction amongst the independent variables and the effect of consecutive irrigant refreshment on percentage of biofilm removal. A Tukey post hoc test was used to evaluate the effect of each independent variable on percentage biofilm removal in the absence of statistically significant interactions. RESULTS: For the lateral canal, NaOCl removed significantly more biofilm irrespective of the number of refreshments and exposure time (P = 0.005). There was no significant effect in biofilm removal between the consecutive irrigant refreshments measured in the same biofilm. For the isthmus, NaOCl removed significantly more biofilm irrespective of the number of refreshments and exposure time; both NaOCl and a high flow rate removed significantly more biofilm when the exposure time was analysed (P = 0.018 and P = 0.029, respectively). Evaluating the effect of consecutive irrigant refreshment on the same biofilm, 2% NaOCl, 0.1 mL s-1 flow rate and one or two refreshments removed significant more biofilm (P = 0.04, 0.034 and 0.003, <0.001, respectively). CONCLUSIONS: In this model, refreshment did not improve biofilm removal from simulated lateral root canal spaces. NaOCl removed more biofilm from the lateral canal- and isthmus-like structure. A higher flow rate removed significantly more biofilm from the isthmus-like structure. There was always remaining biofilm left after the irrigation procedures.

Chemical efficacy of several NaOCl concentrations on biofilms of different architecture: new insights on NaOCl working mechanisms.

AIM: To investigate the anti-biofilm efficacy and working mechanism of several NaOCl concentrations on dual-species biofilms of different architecture as well as the changes induced on the architecture of the remaining biofilms. METHODOLOGY: Streptococcus oralis J22 and Actinomyces naeslundii T14V-J1 were co-cultured under different growth conditions on saliva-coated hydroxyapatite discs. A constant-depth film fermenter (CDFF) was used to grow steady-state, four-day mature biofilms (dense architecture). Biofilms were grown under static conditions for 4 days within a confined space (less dense architecture). Twenty microlitres of buffer, 2-, 5-, and 10% NaOCl were applied statically on the biofilms for 60 s. Biofilm disruption and dissolution, as well as bubble formation, were evaluated with optical coherence tomography (OCT). The viscoelastic profile of the biofilms post-treatment was assessed with low load compression testing (LLCT). The bacteria/extracellular polysaccharide (EPS) content of the biofilms was examined through confocal laser scanning microscopy (CLSM). OCT, LLCT and CLSM data were analysed through one-way analysis of variance (ANOVA) and Tukey's HSD post-hoc test. Linear regression analysis was performed to test the correlation between bubble formation and NaOCl concentration. The level of significance was set at a < 0.05. RESULTS: The experimental hypothesis according to which enhanced biofilm disruption, dissolution and bubble formation were anticipated with increasing NaOCl concentration was generally confirmed in both biofilm types. Distinct differences between the two biofilm types were noted with regard to NaOCl anti-biofilm efficiency as well as the effect that the several NaOCl concentrations had on the viscoelasticity profile and the bacteria/EPS content. Along with the bubble generation patterns observed, these led to the formulation of a concentration and biofilm structure-dependent theory of biofilm removal. CONCLUSIONS: Biofilm architecture seems to be an additional determining factor of the penetration capacity of NaOCl, and consequently of its anti-biofilm efficiency.

Factors affecting the chemical efficacy of 2% sodium hypochlorite against oral steady-state dual-species biofilms: Exposure time and volume application.

AIM: To study the influence of time and volume of 2% sodium hypochlorite (NaOCl) on biofilm removal and to investigate the changes induced on the biofilm architecture. Steady-state, dual-species biofilms of standardized thickness and a realistic contact surface area between biofilms and NaOCl were used. METHODOLOGY: Streptococcus oralis J22 and Actinomyces naeslundii T14V-J1 biofilms were grown on saliva-coated hydroxyapatite discs within sample holders in the Constant Depth Film Fermenter (CDFF) for 96 h. Two per cent NaOCl was statically applied for three different time intervals (60, 120 and 300 s) and in two different volumes (20 and 40 µL) over the biofilm samples. The diffusion-driven effects of time and volume on biofilm disruption and dissolution were assessed with Optical Coherence Tomography (OCT). Structural changes of the biofilms treated with 2% NaOCl were studied with Confocal Laser Scanning Microscopy (CLSM) and Low Load Compression Testing (LLCT). A two-way analysis of variance (2-way anova) was performed, enabling the effect of each independent variable as well as their interaction on the outcome measures. RESULTS: Optical coherence tomography revealed that by increasing the exposure time and volume of 2% NaOCl, both biofilm disruption and dissolution significantly increased. Analysis of the interaction between the two independent variables revealed that by increasing the volume of 2% NaOCl, significant biofilm dissolution could be achieved in less time. Examination of the architecture of the remaining biofilms corroborated the EPS-lytic action of 2% NaOCl, especially when greater volumes were applied. The viscoelastic analysis of the 2% NaOCl-treated biofilms revealed that the preceding application of higher volumes could impact their subsequent removal. CONCLUSIONS: Time and volume of 2% NaOCl application should be taken into account for maximizing the anti-biofilm efficiency of the irrigant and devising targeted disinfecting regimes against remaining biofilms.

Chemical biofilm removal capacity of endodontic irrigants as a function of biofilm structure: optical coherence tomography, confocal microscopy and viscoelasticity determination as integrated assessment tools.

AIM: To investigate the influence of biofilm structure on the biofilm removal capacity of endodontic irrigants and to study changes in the architecture of the remaining biofilms. METHODOLOGY: Streptococcus oralis J22 and Actinomyces naeslundii T14V-J1 were cocultured under different growth conditions on saliva-coated hydroxyapatite discs. A constant depth film fermenter (CDFF) was used to grow steady-state 4-day biofilms. Biofilms were grown under static conditions for 4 and 10 days within a confined space. Twenty microlitres of 2% NaOCl, 2% Chlorhexidine (CHX), 17% Ethylene-diamine-tetra-acetic acid (EDTA) and buffer were applied statically on the biofilms for 60 s. Biofilm removal was evaluated with optical coherence tomography (OCT). Post-treated biofilms were assessed via low load compression testing (LLCT) and Confocal laser scanning microscopy (CLSM). Optical coherence tomography data were analysed through a two-way analysis of variance (ANOVA). Low load compression testing and CLSM data were analysed through one-way ANOVA and Dunnett's post hoc test. The level of significance was set at a < 0.05. RESULTS: The initial biofilm structure affected the biofilm removal capacity of the irrigants. NaOCl demonstrated the greatest chemical efficacy against the biofilms and was significantly more effective on the static than the CDFF biofilms (P < 0.001). CHX was ineffective and caused a rearrangement of the biofilm structure. Ethylene-diamine-tetra-acetic acid exhibited a distinct removal effect only on the CDFF biofilms. Biofilm age influenced the structure of the remaining biofilms. The 4-day grown remaining biofilms had a significantly different viscoelastic pattern compared to the respective 10-day grown biofilms (P ≤ 0.01), especially in the NaOCl-treated group. Confocal laser scanning microscopy analysis confirmed the CHX-induced biofilm structural rearrangement. CONCLUSIONS: Biofilm structure is an influential factor on the chemical efficacy of endodontic irrigants. Optical coherence tomography allows biofilm removal characteristics to be studied. NaOCl should remain the primary irrigant. Ethylene-diamine-tetra-acetic acid was effective against cell-rich/EPS-poor biofilms. Chlorhexidine did not remove biofilm, but rather rearranged its structure.

Cleaning lateral morphological features of the root canal: the role of streaming and cavitation.

AIM: To investigate the effects of ultrasonic activation file type, lateral canal location and irrigant on the removal of a biofilm-mimicking hydrogel from a fabricated lateral canal. Additionally, the amount of cavitation and streaming was quantified for these parameters. METHODOLOGY: An intracanal sonochemical dosimetry method was used to quantify the cavitation generated by an IrriSafe 25 mm length, size 25 file inside a root canal model filled with filtered degassed/saturated water or three different concentrations of NaOCl. Removal of a hydrogel, demonstrated previously to be an appropriate biofilm mimic, was recorded to measure the lateral canal cleaning rate from two different instruments (IrriSafe 25 mm length, size 25 and K 21 mm length, size 15) activated with a P5 Suprasson (Satelec) at power P8.5 in degassed/saturated water or NaOCl. Removal rates were compared for significant differences using nonparametric Kruskal-Wallis and/or Mann-Whitney U-tests. Streaming was measured using high-speed particle imaging velocimetry at 250 kfps, analysing both the oscillatory and steady flow inside the lateral canals. RESULTS: There was no significant difference in amount of cavitation between tap water and oversaturated water (P = 0.538), although more cavitation was observed than in degassed water. The highest cavitation signal was generated with NaOCl solutions (1.0%, 4.5%, 9.0%) (P < 0.007) and increased with concentration (P < 0.014). The IrriSafe file outperformed significantly the K-file in removing hydrogel (P < 0.05). Up to 64% of the total hydrogel volume was removed after 20 s. The IrriSafe file typically outperformed the K-file in generating streaming. The oscillatory velocities were higher inside the lateral canal 3 mm compared to 6 mm from WL and were higher for NaOCl than for saturated water, which in turn was higher than for degassed water. CONCLUSIONS: Measurements of cavitation and acoustic streaming have provided insight into their contribution to cleaning. Significant differences in cleaning, cavitation and streaming were found depending on the file type and size, lateral canal location and irrigant used. In general, the IrriSafe file outperformed the K-file, and NaOCl performed better than the other irrigants tested. The cavitation and streaming measurements revealed that both contributed to hydrogel removal and both play a significant role in root canal cleaning.

[Endodontics in motion: new concepts, materials and techniques 3. The role of irrigants during root canal treatment]. / Endodontologie in beweging: nieuwe concepten, materialen en technieken 3. De rol van activatie van wortelkanaalspoelmiddelen tijdens de wortelkanaalbehandeling.

The aims of root canal irrigation are the chemical dissolution or disruption and the mechanical detachment of pulp tissue, dentin debris and smear layer (instrumentation products), microorganisms (planktonic or biofilm) and their products from the root canal wall, their removal out of the root canal system and their chemical dissolution or disruption. Each of the endodontic irrigation systems has its own irrigant flow characteristics, which should fulfill these aims. Without flow (convection), the irrigant would have to be distributed through diffusion. This process is slow and depends on temperature and concentration gradients. On the other hand, convection is a faster and more efficient transport mechanism. During irrigant flow, frictional forces will occur, for example between the irrigant and the root canal wall (wall shear stress). These frictional forces have a mechanical cleaning effect on the root canal wall. These frictional forces are the result of the flow characteristics related to the different irrigation systems.

Influence of refreshment/activation cycles and temperature rise on the reaction rate of sodium hypochlorite with bovine dentine during ultrasonic activated irrigation.

AIM: To evaluate the effect of multiple refreshment/activation cycles and temperature on the reaction rate of sodium hypochlorite (NaOCl) with bovine dentine during ultrasonic activated irrigation (UAI) under laboratory conditions. METHODOLOGY: The root canal walls of 24 standardized root canals in bovine incisors were exposed to a standardized volume of NaOCl at different temperatures (24 °C and 38 °C) and exposure times (20, 60 and 180 s). The irrigant was refreshed and ultrasonically activated four times for 20 s followed by a 40 s rest interval, with no refreshment and no activation as the controls. The reaction rate was determined by measuring the amount of active chlorine in the NaOCl solution before and after being exposed to dentine during the specific experimental conditions. Calorimetry was used to measure the electrical-to-sonochemical conversion efficiency during ultrasonic activation. RESULTS: Refreshment, activation and exposure time all increased the reaction rate of NaOCl (P < 0.05). During activation, the temperature of the irrigant increased up to 10 °C. Such temperature rise was insufficient to enhance the reaction rate of NaOCl (P > 0.125). CONCLUSIONS: The reaction rate of NaOCl with dentine is enhanced by refreshment, ultrasonic activation and exposure time. Temperature rise of irrigant during ultrasonic activation was not sufficient to alter the reaction rate.

Formation and removal of apical vapor lock during syringe irrigation: a combined experimental and Computational Fluid Dynamics approach.

AIM: (i) To evaluate the effect of needle type and insertion depth, root canal size and irrigant flow rate on the entrapment of air bubbles in the apical part of a root canal (apical vapor lock) during syringe irrigation using experiments and a Computational Fluid Dynamics (CFD) model, (ii) to investigate whether the irrigant contact angle affects bubble entrapment, (iii) to examine if an established vapor lock can be removed by syringe irrigation. METHODOLOGY: Bubble entrapment during irrigation of straight artificial root canals of size 35 or 50 was evaluated by real-time visualizations. The irrigant was delivered by a closed-ended or an open-ended needle positioned at 1 or 3 mm short of working length (WL) and at a flow rate of 0.033-0.260 mL s(-1) . Results were analysed by nonparametric tests at 0.05 significance. Selected cases were also simulated by a two-phase CFD model. RESULTS: A vapor lock was observed in 48% of the cases investigated experimentally. Increasing the apical size, using an open-ended needle, positioning the needle closer to WL and delivering the irrigant at higher flow rate resulted in significantly smaller vapor lock. An increased contact angle resulted in the entrapment of a larger bubble when a low flow rate was used. Both brief insertion of the needle to WL whilst irrigating at a flow rate of 0.083 mL s(-1) and delivering the irrigant at 0.260 mL s(-1) without changing the needle position were capable of removing an established vapor lock. CONCLUSIONS: Apical vapor lock may occur under certain conditions, but appears to be easily prevented or removed by syringe irrigation.

A novel methodology providing insights into removal of biofilm-mimicking hydrogel from lateral morphological features of the root canal during irrigation procedures.

AIM: To introduce and characterize a reproducible hydrogel as a suitable biofilm mimic in endodontic research. To monitor and visualize the removal of hydrogel from a simulated lateral canal and isthmus for the following: I) Ultrasonic-Activated Irrigation (UAI) with water, ii) UAI with NaOCl and iii) NaOCl without UAI. METHODOLOGY: A rheometer was used to characterize the viscoelastic properties and cohesive strength of the hydrogel for suitability as a biofilm mimic. The removal rate of the hydrogel from a simulated lateral canal or isthmus was measured by high-speed imaging operating at frame rates from 50 to 30,000 fps. RESULTS: The hydrogel demonstrated viscoelastic behaviour with mechanical properties comparable to real biofilms. UAI enhanced the cleaning effect of NaOCl in isthmi (P < 0.001) and both NaOCl and water in lateral canals (P < 0.001). A greater depth of cleaning was achieved from an isthmus (P = 0.009) than from a lateral canal with UAI and also at a faster rate for the first 20 s. NaOCl without UAI resulted in a greater depth of hydrogel removal from a lateral canal than an isthmus (P < 0.001). The effect of UAI was reduced when stable bubbles were formed and trapped in the lateral canal. Different removal characteristics were observed in the isthmus and the lateral canal, with initial highly unstable behaviour followed by slower viscous removal inside the isthmus. CONCLUSIONS: The biofilm-mimicking hydrogel is reproducible, homogenous and can be easily applied and modified. Visualization of its removal from lateral canal anatomy provides insights into the cleaning mechanisms of UAI for a biofilm-like material. Initial results showed that UAI improves hydrogel removal from the accessory canal anatomy, but the creation of stable bubbles on the hydrogel-liquid interface may reduce the cleaning rate.

Acoustic streaming induced by an ultrasonically oscillating endodontic file.

Ultrasonically activated irrigation is an advanced dental technique for irrigation of the root canal system during a root canal treatment. The basic cleaning mechanism is a result of acoustic streaming induced by an oscillating file, leading to mixing of the irrigant and pressure and shear stresses on the walls of the root canal. Here the induced acoustic streaming, pressure, and shear stress are investigated in a two-dimensional cross-section of the root canal, using a combination of theory, numerical predictions, and experimental validation through high-speed particle tracking velocimetry. Acoustic streaming theory describes very well the flow induced by an ultrasonically oscillating endodontic file. It consists of an oscillatory component, which is dominant near the file, and a steady component, or jet, along the axis of oscillation. The importance of the oscillatory component for both the pressure and the shear stress is apparent, as it is two to three orders of magnitude higher than the steady component. A confinement affects the formation of the steady jets; meanwhile the oscillatory velocities and associated pressure and shear stress are increased. Previous work considering only the steady component of the flow therefore, underestimated the hydrodynamic effects induced by ultrasonic files.

Factors affecting irrigant extrusion during root canal irrigation: a systematic review.

The aim of the present study was to conduct a systematic review and critical analysis of published data on irrigant extrusion to identify factors causing, affecting or predisposing to irrigant extrusion during root canal irrigation of human mature permanent teeth. An electronic search was conducted in Cochrane Library, LILACS, PubMed, SciELO, Scopus and Web of Knowledge using a combination of the terms 'irrigant', 'rinse', 'extrusion', 'injection', 'complication', 'accident', 'iatrogenic', 'root canal', 'tooth' and 'endodontic'. Additional studies were identified by hand-searching of six endodontic journals and the relevant chapters of four endodontic textbooks, resulting in a total of 460 titles. No language restriction was imposed. After applying screening and strict eligibility criteria by two independent reviewers, 40 case reports and 10 ex vivo studies were included in the review. A lack of clinical studies focusing on irrigant extrusion during root canal irrigation was evident. The reviewed case reports focused mainly on the clinical manifestations and management of the accidents and did not provide adequate details on the possible factors that may influence irrigant extrusion. The data from the included ex vivo studies were inconclusive due to major methodological limitations, such as not simulating the presence of periapical tissues and not assessing the validity of irrigant detection methods. The extensive variability in the protocols employed hindered quantitative synthesis. The choice of factors investigated in ex vivo studies seems not to have been driven by the available clinical evidence. These issues need to be addressed in future studies.

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

AIM: (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. METHODOLOGY: 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. RESULTS: 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. 5 kHz), but still performed the original 30 kHz oscillations. Travelling waves were identified on the file. The file oscillation was not dampened completely due to the contact and hydrodynamic cavitation was detected. CONCLUSION: Considerable file-to-wall contact occur-red during irrigant activation. Therefore, the term 'Passive Ultrasonic Irrigation' should be amended to 'Ultrasonically Activated Irrigation'.

The effects of hyperosmosis or high pH on a dual-species biofilm of Enterococcus faecalis and Pseudomonas aeruginosa: an in vitro study.

AIM: To investigate the effect of hyperosmotic hyperosmosis or alkaline stress on a dual-species biofilm of Enterococcus faecalis and Pseudomonas aeruginosa. METHODOLOGY: Biofilms were grown on glass cover slips suspended in bacterial inoculate for 96 h, after which the cover slips with attached biofilms were immersed in brain heart infusion broth (BHI-broth) with 6 mol L(-1) sodium chloride (NaCl) representing the hyperosmotic group or Ca(OH)(2), pH 12.1, representing the alkaline group. Two per cent sodium hypochlorite and BHI- broth served as positive and negative controls, respectively. After treatment, the biofilms were washed, harvested and plated on blood-agar plates after serial dilution. The bactericidal effect was assessed by determining the colony-forming units (CFU). The effect on the biofilm mass was imaged with confocal laser scanning microscopy (CLSM). RESULTS: Hyperosmosis reduced the CFU of both species significantly after 72 h (P < 0.0001). After 168 h, P. aeruginosa was eradicated and the E. faecalis reduction was more than 99%. High pH could not induce a significant bacterial reduction. CLSM revealed dense flocculation of the biofilms incubated in alkaline broth. CONCLUSION: Hyperosmosis effectively reduced a dual-species biofilm of E. faecalis and P. aeruginosa, whilst high pH had limited bactericidal effect in this model.

Irrigant flow in the root canal: experimental validation of an unsteady Computational Fluid Dynamics model using high-speed imaging.

AIM: To compare the results of a Computational Fluid Dynamics (CFD) simulation of the irrigant flow within a prepared root canal, during final irrigation with a syringe and a needle, with experimental high-speed visualizations and theoretical calculations of an identical geometry and to evaluate the effect of off-centre positioning of the needle inside the root canal. METHODOLOGY: A CFD model was created to simulate irrigant flow from a side-vented needle inside a prepared root canal. Calculations were carried out for four different positions of the needle inside a prepared root canal. An identical root canal model was made from poly-dimethyl-siloxane (PDMS). High-speed imaging of the flow seeded with particles and Particle Image Velocimetry (PIV) were combined to obtain the velocity field inside the root canal experimentally. Computational, theoretical and experimental results were compared to assess the validity of the computational model. RESULTS: Comparison between CFD computations and experiments revealed good agreement in the velocity magnitude and vortex location and size. Small lateral displacements of the needle inside the canal had a limited effect on the flow field. CONCLUSIONS: High-speed imaging experiments together with PIV of the flow inside a simulated root canal showed a good agreement with the CFD model, even though the flow was unsteady. Therefore, the CFD model is able to predict reliably the flow in similar domains.

Reaction rate of NaOCl in contact with bovine dentine: effect of activation, exposure time, concentration and pH.

AIM: To determine the influence of activation method (ultrasound or laser), concentration, pH and exposure time on the reaction rate (RR) of NaOCl when in contact with dentinal walls. METHODOLOGY: The walls from standardized root canals in bovine incisors were exposed to a standardized volume of sodium hypochlorite (NaOCl) with different concentrations (2% and 10%), pH (5 and 12) and exposure times (1 and 4min). Two irrigation protocols were tested: passive ultrasonic irrigation or laser activated irrigation with no activation as the control. The activation interval lasted 1min followed by a rest interval of 3 min with no activation. The RR was determined by measuring the iodine concentration using an iodine/thiosulfate titration method. RESULTS: Exposure time, concentration and activation method influenced the reaction rate of NaOCl whereas pH did not. CONCLUSIONS: Activation is a strong modulator of the reaction rate of NaOCl. During the rest interval of 3min, the consumption of available chlorine increased significantly. This effect seems to be more pronounced after irrigant activation by laser. pH did not affect the reaction rate of 2% NaOCl.

The effect of root canal taper on the irrigant flow: evaluation using an unsteady Computational Fluid Dynamics model.

AIM: To evaluate the effect of root canal taper on irrigant flow inside a prepared root canal during final irrigation with a syringe and two types of needles, using a Computational Fluid Dynamics (CFD) model. METHODOLOGY: A validated CFD model was used to simulate irrigant flow from either a side-vented or a flat 30G needle positioned inside size 30, .02 taper, 30, .04, 30, .06, ProTaper F3 or size 60, .02 taper root canals, at 3 mm short of working length (WL). Velocity, pressure and shear stress in the root canal were evaluated. RESULTS: The side-vented needle could not achieve irrigant replacement to the WL in any of the cases. Significant irrigant replacement was evident further than 2 mm apically to the tip of the flat needle in the size 30, .06 taper, F3 and size 60, .02 taper canal. A wider distribution of wall shear stress was noted as the canal taper increased but the maximum shear stress decreased. The flat needle led to higher mean pressure at the apical foramen. Both needles showed a similar gradual decrease in apical pressure as the taper increased, but the least pressure was calculated in the size 60, .02 taper canal. CONCLUSIONS: An increase in root canal taper improved irrigant replacement and wall shear stress whilst reducing the risk for irrigant extrusion. Irrigant flow in a minimally tapered root canal with a large apical preparation size also improved irrigant replacement and wall shear stress and reduced the risk for irrigant extrusion, compared to the tapered root canals with a smaller apical preparation size.

The effect of apical preparation size on irrigant flow in root canals evaluated using an unsteady Computational Fluid Dynamics model.

AIM: To evaluate the effect of apical preparation size on irrigant flow inside a root canal during final irrigation with a syringe and two different needles types, using a Computational Fluid Dynamics (CFD) model. METHODOLOGY: A validated CFD model was used to simulate the irrigant flow from either a side-vented or a flat 30G needle positioned inside root canals having sizes of 25, 35, 45 and 55, all with a .06 taper, at 3 mm short of working length (WL). Velocity, pressure and shear stress in the root canal were evaluated. RESULTS: Different preparation sizes resulted in minor differences in the flow pattern in the apical root canal. Major differences were observed between the two needle types. The side-vented needle could not achieve irrigant replacement to the WL even in a size 55, .06 taper root canal. Significant irrigant replacement was evident almost to the WL in size 35, 45 and 55, .06 taper root canals with the flat needle. The maximum shear stress decreased as the preparation size increased. The flat needle developed higher mean pressure at the apical foramen. Both needles led to a similar gradual decrease in apical pressure as the preparation size increased. CONCLUSIONS: Apical preparation size affected irrigant replacement, the shear stress on the canal wall and the pressure at the apical foramen. Root canal enlargement to sizes larger than 25 appeared to improve the performance of syringe irrigation. Adequate space between the needle and the canal wall should be ensured to allow for an effective reverse flow of the irrigant towards the canal orifice.