Evaluation of the antifungal activity of four solutions used as a final rinse in vitro

The aim of the study was to compare the antifungal activity of 1.3% sodium hypochlorite (NaOCl), 2% chlorhexidine (CHX), MTAD and Tetraclean as a final rinse against Candida albicans in a human tooth model in vitro. Ninety extracted human maxillary central and lateral incisor teeth were randomly divided into four groups each with 20 teeth, a positive and a negative control each with five teeth. After preparing the root canals, teeth were inoculated with Candida albicans (ATCC 10261) and incubated for 72 h. Teeth were divided into four experimental groups according to the irrigation solution as follows: NaOCl, CHX, MTAD and Tetraclean. After culturing aliquots from the experimental teeth on Sabouraud 4% dextrose agar, colony‐forming units were counted. The results showed that 1.3% NaOCl and 2% CHX were equally effective and significantly superior to MTAD and Tetraclean (P < 0.05). Furthermore, antifungal efficacy of Tetraclean was significantly superior to MTAD (P < 0.05).     

Residual antibacterial activity of chlorhexidine and MTAD in human root dentin in vitro

The purpose of this in vitro study was to compare the antimicrobial substantivity of BioPure MTAD, 2% chlorhexidine (CHX) and 2.6% sodium hypochlorite (NaOCl) in human root dentin. One hundred and ten dentin tubes prepared from human maxillary incisors were infected in vitro for 14 days with Enterococcus faecalis. The specimens were divided into five groups as follows: CHX; BioPure MTAD; NaOCl; infected dentin tubes (positive control); and sterile dentin tubes (negative control). Dentin chips were collected with round burs into Brain Heart Infusion (BHI) broth. After culturing, the number of colony-forming units (CFU) was counted. In all experimental groups, CFU was minimum after treatment (day 0), and the results obtained were significantly different from each other at any time period (P < 0.05). After treatment, the NaOCI group and BioPure MTAD group showed the lowest and highest number of CFU, respectively. In each group, the number of CFUs increased significantly by time-lapse (P < 0.05). In conclusion, the substantivity of BioPure MTAD was significantly greater than CHX and NaOCl.     

Comparative evaluation of antimicrobial efficacy of sodium hypochlorite, MTAD, and Tetraclean against Enterococcus faecalis biofilm

The aim of this study was to compare the antimicrobial efficacy of 5.25% NaOCl, BioPure MTAD (Dentsply Tulsa Dental, Johnson City, TN), and Tetraclean (Ogna Laboratori Farmaceutici, Milano, Italy) against Enterococcus faecalis biofilm generated on cellulose nitrate membrane filters. After incubation, the membrane filters were transferred into tubes containing 5 mL of the selected antimicrobial solution test agent or NaCl 0.9% (positive control) and incubated for 5, 30, and 60 minutes at 20 degrees C. After each period of time, the test agents were vortexed for 60 seconds to resuspend the microorganisms. Ten-fold serial dilutions were generated in reduced transport fluid. Each dilution was plated onto a brain heart infusion plates. The plates were then incubated for 48 hours in an aerobic atmosphere at 37 degrees C and colony-forming units per membrane was calculated. Statistical analysis showed that only 5.25% NaOCl can disgregate and remove the biofilm at every time; however, treatment with Tetraclean caused a high degree of biofilm disgregation in every considered time intervals as compared with MTAD (T5 p < 0.05, T30 p < 0.01, and T60 p < 0.001).     

Effect of MTAD on flexural strength and modulus of elasticity of dentin

The purpose of this study was to evaluate the effect of MTAD on the flexural strength and modulus of elasticity of dentin. Dentin bars were randomly assigned to eight groups treated with various concentrations of NaOCl (5.25%, 2.65%, 1.31%, 0.66%), 17% EDTA, MTAD, saline, or according to a clinical protocol with 1.3% NaOCl and then 5 min of MTAD. Specimens were subjected to flexural strength and modulus of elasticity tests. Two-sample t test (alpha = 0.05) showed no statistically significant differences between various groups except for a reduction in flexural strength for the 2-h MTAD group (p = 0.002) and the EDTA group (p = 0.002). A significant reduction of modulus of elasticity for the 2-h MTAD group (p < 0.001), EDTA group (p < 0.001), and 0.6% NaOCl (p < 0.002) also was noted. There was no significant difference in flexural strength and modulus of elasticity between the dentin bars exposed to saline or MTAD when applied according to the clinical protocol (p > 0.05). These results indicate that MTAD can be used as prescribed for clinical use without affecting the physical properties of the dentin.     

Quantitative analysis of the solubilizing action of MTAD,sodium hypochlorite,and EDTA on bovine pulp and dentin

Necrotic bovine pulp and dentin were used in this study as model tissues to represent the organic and inorganic components of the smear layer present in instrumented root canals. The capabilities of endodontic irrigants to dissolve pulverized forms of these tissues were compared. Lyophilized tissue samples were mixed for 2 h at 37 degrees C with MTAD, three concentrations of sodium hypochlorite (NaOCl), 17% EDTA, or isotonic saline. Undissolved tissues were rinsed with water and lyophilized. The change in tissue weight after exposure to an irrigant was measured to quantify solubilization. The results showed that various concentrations of NaOCl removed organic components of pulp and dentin effectively. As pulp solubilizers 5.25% and 2.60% NaOCl were equal (>90%), and 5.25% NaOCl was capable of dissolving virtually the entire organic component of dentin. EDTA was capable of solubilizing inorganic material in dentin and organic material in pulp and likely also in dentin. It dissolved >70% of the dentin and >51% of the pulp. The solubilizing effects of MTAD on pulp and dentin were somewhat similar to those of EDTA. The major difference between the actions of these solutions was a high binding affinity of doxycycline present in MTAD for the dentin.     

Effects of reducing agents on birefringence dentin collagen after use of different endodontic auxiliary chemical substances

Introduction

The aim of this study was to evaluate the effect of 10% ascorbic acid or 10% sodium ascorbate on organic matrix collagen of bovine dentin root canal walls after irrigation with 5.25% sodium hypochlorite (NaOCl), 17% ethylenediaminetetraacetic acid (EDTA), or 0.9% sodium chloride.

Methods

Eighty bovine incisors were randomly divided into 8 groups (n = 10): group 1, 0.9% sodium chloride (control); group 2, 5.25% NaOCl + 17% EDTA (NaOCl + EDTA); group 3, 5.25% NaOCl + 17% EDTA + 10% ascorbic acid (NaOCl + EDTA + AA); group 4, 5.25% NaOCl + 17% EDTA + 10% sodium ascorbate (NaOCl + EDTA + SA); group 5, 5.25% NaOCl (NaOCl); group 6, 17% EDTA; group 7, 10% ascorbic acid (AA); and group 8, 10% sodium ascorbate (SA). Teeth were chemomechanically prepared, submitted to histologic processing, and stained with Sirius Red dye to be analyzed under polarized light microscopy. Absorbance assay was also performed to confirm the loss of collagen.

Results

NaOCl + EDTA and NaOCl groups presented a significantly different birefringence pattern compared with the control group (P < .05). The measurement of the optical retardations of NaOCl + EDTA + SA indicated that this group was not statistically different from the control group. Although the measurement of the optical retardations of NaOCl + EDTA + AA was statistically different from the control group, the results were significantly higher than for NaOCl + EDTA. The birefringence of EDTA, AA, and SA groups was not statistically different from that of control group. The absorbance assay of NaOCl + EDTA and NaOCl groups confirmed the loss of collagen (P < .05).

Conclusions

It is possible to conclude that 5.25% NaOCl, whether associated or not with 17% EDTA, causes birefringence alterations and loss of dentin collagen. These alterations reduced the ability of Sirius Red to bind with collagen fiber molecules. The reductions in the optical retardation values could be reversed by the application of either 10% ascorbic acid or 10% sodium ascorbate after 5.25% sodium hypochlorite and 17% EDTA irrigation.     

A comparison of the antimicrobial efficacy of NaOCl/Biopure MTAD versus NaOCl/EDTA against Enterococcus faecalis

The purpose of this investigation was to compare the antimicrobial efficacy of irrigating with 1.3% NaOCl/Biopure MTAD versus irrigation with 5.25% NaOCl/15% EDTA in the apical 5 mm of roots infected with Enterococcus faecalis. Bilaterally matched human teeth were sterilized and inoculated with E. faecalis. After chemomechanical root canal preparation, the root-ends were resected and pulverized in liquid nitrogen to expose E. faecalis in dentinal tubules or other recesses away from the main root canal system. The number of colony forming units (CFU) of E. faecalis per mg was determined from the pulverized root-ends. No significant differences were seen (t = 0.70, p = 0.495) between the number of colony forming units of E. faecalis for teeth irrigated with 5.25% NaOCl/15% EDTA (mean 131 +/- 291 CFU/mg) versus those teeth irrigated with 1.3% NaOCl/Biopure MTAD (mean 187 +/- 237 CFU/mg). This study demonstrated that there is no difference in antimicrobial efficacy for irrigation with 5.25% NaOCl/15% EDTA versus irrigation with 1.3% NaOCl/Biopure MTAD in the apical 5 mm of roots infected with E. faecalis.     

Effect of endodontic irrigants on the shear bond strength of epiphany sealer to dentin

This study examined the effects of commonly employed endodontic irrigants on Epiphany-dentin bond strengths. Smear layers were created on dentin discs obtained from human third molars. Five groups of 10 discs each were treated with one of the following: (a) water; (b) 2% chlorhexidine; (c) 6% sodium hypochlorite (NaOCl); (d) 6% NaOCl followed by EDTA and water; or (e) 1.3% NaOCl followed by MTAD. The treated surfaces were air-dried and treated with Epiphany primer. Hollow steel tubes were placed on the dentin surfaces, and Epiphany sealer was applied to dentin inside the tubes. Samples were tested for shear bond strengths after 7 days of storage. Representative specimens were examined using scanning electron microscopy. One-way ANOVA and Tukey's test revealed significant differences among the groups (p<0.05). Using water or chlorhexidine as an irrigant resulted in significantly lower bond strengths when compared with NaOCl, NaOCl/EDTA, or NaOCl/MTAD. Neither EDTA nor MTAD significantly improved Epiphany-dentin bond strengths when compared with NaOCl used alone.     

The antimicrobial effect of new and conventional endodontic irrigants on intra-orally infected dentin

Abstract

Objectives. To evaluate if the incorporation of antimicrobial compounds to chelating agents or the use of chelating agents with antimicrobial activity as 7% maleic acid and peracetic acid show similar disinfection ability in comparison to conventional irrigants as sodium hypochlorite or iodine potassium iodide against biofilms developed on dentin. Materials and methods. The total bio-volume of live cells, the ratio of live cells and the substratum coverage of dentin infected intra-orally and treated with the irrigant solutions: MTAD, Qmix, Smear Clear, 7% maleic acid, 2% iodine potassium iodide, 4% peracetic acid, 2.5% and 5.25% sodium hypochlorite was measured by using confocal microscopy and the live/dead technique. Five samples were used for each irrigant solution. Results. Several endodontic irrigants containing antimicrobials as clorhexidine (Qmix), cetrimide (Smear Clear), maleic acid, iodine compounds or antibiotics (MTAD) lacked an effective antibiofilm activity when the dentin was infected intra-orally. The irrigant solutions 4% peracetic acid and 2.5–5.25% sodium hypochlorite decrease significantly the number of live bacteria in biofilms, providing also cleaner dentin surfaces (p < 0.05). Conclusions. Several chelating agents containing antimicrobials could not remove nor kill significantly biofilms developed on intra-orally infected dentin, with the exception of sodium hypochlorite and 4% peracetic acid. Dissolution ability is mandatory for an appropriate eradication of biofilms attached to dentin.     

Time-dependent Decalcifying Effects of Endodontic Irrigants with Antibacterial Properties

The aim of this study was to investigate the extent of calcium ion (Ca²⁺) removal on root dentin after treatment with 1.25% NaOCl, 2.5% NaOCl, 5.25% NaOCl, 2% chlorhexidine (CHX), 2% iodine potassium iodide (IKI), and MTAD in 3 immersion time periods. Extracted human mandibular premolars were bisected longitudinally, and the root halves (n = 70) were isolated with nail varnish, leaving the root canal exposed. The specimens were immersed in the test solutions, during which the amount of Ca²⁺ release into the solutions was determined at 5, 10, and 15 minutes by flame photometry. The decalcifying effect of 5%, 2.5%, and 1.25% NaOCl and 2% IKI significantly increased within time (P < .05), with 5% NaOCl extracting the greatest amount of Ca²⁺ at all treatment times. For 2% CHX and distilled water, a significant increase in the amount of Ca²⁺ extraction from root dentin was only evident at 10 minutes (P < .05) and did not change at 15-minute readings. Between the 5- to 10-minute and 10- to 15-minute intervals, the greatest amount of increase in the rate of Ca²⁺ extraction from root dentin was observed in the 2.5% NaOCl group. At 15 minutes, 2% CHX and distilled water showed the least amount of change. Among the test solutions, MTAD extracted the least amount of Ca²⁺ at 5 minutes.     

The antimicrobial effect of MTAD: an in vitro investigation

Pulp and periradicular diseases are of microbial origin. To effectively clean the root canal system a disinfecting agent must be able to penetrate into difficult-to-reach areas and kill microorganisms with minimal damage to the host tissues. The purpose of this investigation was to test the ability of a mixture of a tetracycline isomer, an acid, and a detergent (MTAD) to kill Enterococcus faecalis and compare its efficacy to that of sodium hypochlorite (NaOCl) and ethylene diamine tetraacetic acid (EDTA). The zones of inhibition and minimum inhibitory concentrations were measured for these solutions. Measurement of zones of inhibition and determination of the minimum inhibitory concentrations showed that MTAD is as effective as 5.25% NaOCl and significantly more effective than EDTA (p < 0.0001). Furthermore, MTAD is significantly more effective in killing E. faecalis than NaOCl when the solutions are diluted (p < 0.0001). Measurement of the minimum inhibitory concentrations demonstrated that although MTAD is still effective in killing E. faecalis at 200x dilution, NaOCl ceases to exert its antibacterial activity beyond 32x dilution. EDTA did not exhibit any antibacterial activity. Based on the results of this study, it seems that MTAD is an effective solution in eradicating E. faecalis.     

The effects of calcium hydroxide removal methods on bond strength of Epiphany SE with two irrigation protocols

Abstract

Objective: The purpose of this study was to compare the effects of five calcium hydroxide removal methods on the bond strength of Epiphany SE sealer after canal irrigation with NaOCl+EDTA or NaOCl+MTAD. Materials and methods: The 120 roots were instrumented by using the ProTaper rotary system under irrigation with 2.5% sodium hypochlorite and randomly divided into two major groups according to the final irrigation: 1.3% NaOCl + MTAD and 5% NaOCl + 17% EDTA. For controls, 10 roots from each of the final irrigations with NaOCl + MTAD and NaOCl + EDTA (20 roots) were not filled with Ca(OH)₂. The intra-canal paste, Ca(OH)₂ was applied to each of the 100 remaining roots and stored at 37°C for 7 days. Each group was sub-divided into five sub-groups (n = 10) according to the removal technique for the intra-canal dressing: Group-1: recapitulation with size 30 K file + 3 ml of saline solution, Group-2: recapitulation with size 30 K file + 3 ml of 5% NaOCl, Group-3: using 5% NaOCl + 17% EDTA, Group-4: using 5%NaOCl + 15% citric acid, and Group-5: using 1.3% NaOCl + 5 ml MTAD. The root canals were filled with Resilon and Epiphany SE sealer. The bond strength was measured. Results: The mean bond strength of Epiphany SE to root dentine irrigated with NaOCl + MTAD was lower than that of NaOCl + EDTA. The highest bond strengths were 3.31 ± 0.057 and 2.60 ± 0.054 in the NaOCl + citric acid group when Ca(OH)₂ was applied to roots treated with NaOCl + EDTA and NaOCl + MTAD, respectively (p < 0.05). Conclusion: For root canals treated with NaOCl + EDTA or NaOCl + MTAD, the use of NaOCl + chelating agent for Ca(OH)₂ removal does not adversely affect the bond strength of Epiphany SE to dentin.     

Effect of dentin treatment time with tetraclean on its residual antibacterial activity

The goal of this study was to evaluate the effect of dentin treatment duration (10 minutes, 24 hours, and seven days) with Tetraclean on its residual antibacterial activity in bovine root dentin. Results showed that the number of colony-forming units in all three experimental groups was zero at the first culture. Furthermore, the 10-minute group and seven-day group demonstrated the highest and the lowest number of colony-forming units, respectively.     

Quantitative analysis of the effect of irrigant solution sequences on dentin erosion

Objective

The purpose of the study was to examine the level of erosion on root canal wall dentin caused by immersion in different irrigant solutions in alternative sequences.

Methods

Dentin specimens from teeth with one root canal were instrumented and randomly divided into five groups. Each group was subjected to 17% EDTA, 17% ethyleneglycoltetraacetic acid (EGTA), or 10% citric acid (CA) and 5.25% sodium hypochlorite (NaOCl) varying the time of irrigant exposure and the order of the irrigants. The specimens were examined by scanning electron microscopy (SEM), and randomized digital images of the dentin surface were taken. The area of tubule openings was measured by a semiautomatic method.

Results

No erosion was detected when demineralizing agents were used as a final rinse after NaOCl. However, the erosion of peritubular and intertubular dentin was detected when EDTA, EGTA, or CA were used first followed by 5.25% NaOCl (P < .05), and an increase over 100% in the area of dentin tubule openings was measured (P < .01).

Conclusions

NaOCl used as a final irrigant solution after demineralization agents causes marked erosion of root canal dentin.     

Effects of different irrigation solutions on root dentine microhardness,smear layer removal and erosion

This study aimed to compare the effects of different irrigants on root dentine microhardness, erosion and smear layer removal. A total of 72 root dentine slices were divided into six groups, according to the final irrigants used: Group 1: 17% ethylenediamine tetra‐acetic acid (EDTA) + 2.5% NaOCl, Group 2: 7% maleic acid (MA) + 2.5% sodium hypochloride (NaOCl), Group 3: 1.3% NaOCl + mixture of tetracycline, acid and detergent (MTAD), Group 4: Smear Clear + 2.5% NaOCl, Group 5: 5% NaOCl, Group 6: saline. Vickers microhardness values were measured before and after treatment. In total, 42 root‐halves were prepared for scanning electron microscope to evaluate the amount of smear and erosion in the coronal, middle and apical thirds. Data were analysed using two‐way anova , Duncan and two‐proportion z‐tests. Maleic acid showed the greatest reduction in dentine microhardness (P < 0.05), followed by EDTA and MTAD. EDTA, maleic acid, MTAD and Smear Clear removed smear layer efficiently in the coronal and middle thirds of root canal. However, in the apical region, maleic acid showed more efficient removal of the smear layer than the other irrigants (P < 0.05).     

Evaluation of the antibacterial substantivity of several intra-canal agents

The aim of this in vitro study was to compare the antimicrobial substantivity of 2% chlorhexidine gluconate (CHX), 100 mg ml(-1) doxycycline and 2.6% sodium hypochlorite (NaOCl) in bovine root dentine. Eighty dentine tubes prepared from bovine incisors were infected in vitro for 14 days with Enterococcus faecalis. The specimens were divided into five groups as follows: doxycycline HCl; CHX; NaOCl; infected dentine tubes (positive control); and sterile dentine tubes (negative control). Dentine chips were collected with round burs into tryptic soy broth. After culturing, the number of colony-forming units (CFU) was counted. In all experimental groups, the number of CFU was minimum in the first cultures, and the results obtained were significantly different from each other at any time period (P < 0.05). In the first culture, the NaOCl group and doxycycline HCl group showed the lowest and highest number of CFU, respectively. In each group, the number of CFU increased significantly by time-lapse (P < 0.05). In conclusion, the substantivity of CHX was significantly greater than NaOCl and doxycycline.     

Effect of MTAD on Enterococcus faecalis-contaminated root canals of extracted human teeth

The purpose of this investigation was to compare the antimicrobial effect of MTAD (a mixture of a tetracycline isomer, an acid, and a detergent) with that of NaOCl with and without EDTA. Eighty-five extracted human teeth were contaminated with Enterococcus faecalis for 4 weeks. After biomechanical instrumentation using 1.3% or 5.25% NaOCl as root canal irrigant, the root canal and the external surface of each tooth were exposed to a 5-min application of MTAD, 1.3% NaOCl, 5.25% NaOCl or a 1-min application of EDTA followed by irrigation with 5 ml of 1.3% NaOCl or 5.25% NaOCl. Teeth or dentin shavings were cultured to determine presence or absence of the test bacteria. Fisher's exact test showed that the combination of 1.3% NaOCl as a root canal irrigant and MTAD as a final rinse was significantly more effective against E. faecalis than the other regimens. The chi2 test showed no difference between the other regimens.     

Effect of MTAD on the bond strength to enamel and dentin

The purpose of this study was to compare the effect of MTAD (a mixture of a tetracycline isomer [doxycycline], an acid [citric acid], and a detergent [Tween 80]) and phosphoric acid on the bond strength to enamel and dentin using a conventional OptiBond Solo Plus dentin adhesive system. One hundred flat surfaces (50 enamel/50 dentin) were prepared for bonding. Specimens were embedded in acrylic with the flat surface exposed. One of the following surface treatments was applied before bonding composite with Optibond Solo Plus (Kerr Corp): 1 min NaOCl/1 min EDTA; 1 min NaOCl/1 min MTAD; 30 s etch with H3PO4 (positive control), 2 min saline (negative control), or 20 min NaOCl/5 min MTAD (clinical protocol for MTAD). Shear bond strength was tested 30 min after bonding. Data were analyzed using a one-way ANOVA followed by the Student-Newman-Keuls multiple comparison method. Surface pretreatment with acid etch resulted in the greatest enamel bond strengths (p < 0.05). All surface pretreatments were superior to saline for dentinal bonding (p < 0.05). On the basis of our findings, it appears that teeth endodontically treated with the MTAD protocol for clinical use (20 min 1.3% NaOCl/5 min MTAD) may not need any additional dentin conditioning before the application of the dental adhesive.     

Endodontic Chelators Induce Nitric Oxide Expression by Murine-cultured Macrophages

IntroductionEndodontic chelators may extrude to apical tissues during instrumentation activating cellular events on periapical tissues. This study assessed in vitro the expression of nitric oxide (NO) concentrations by murine peritoneal macrophages after contact with MTAD (Dentsply/Tulsa, Tulsa, OK), Tetraclean (Ogna Laboratori Farmaceutici, Muggio, Italy), Smear Clear (Sybron Endo, Orange, CA), and EDTA (Biodinâmica, Ibiporã, PR, Brazil).MethodsMacrophage cells were obtained from Swiss mice after peritoneal lavage. Chelators were diluted in distilled water obtaining 12 concentrations, and MTT assay identified the concentrations, per group, displaying the highest cell viability (analysis of variance, p < 0.01). Selected concentrations were tested for NO expression using Griess reaction. Culture medium and lipopolysaccharide (LPS) were used as controls.ResultsAnalysis of variance and Tukey tests showed that all chelators displayed elevated NO concentrations compared with the negative control (p < 0.01). MTAD induced the lowest NO expression, followed by Tetraclean, EDTA, and Smear Clear. No difference was observed between MTAD and Tetraclean (p > 0.01), Tetraclean and EDTA (p > 0.01), and EDTA and Smear Clear (p > 0.01). LPS ranked similar to both EDTA and Smear Clear (p > 0.01).ConclusionThe tested endodontic chelators displayed severe proinflammatory effects on murine-cultured macrophages. Citric acid–based solutions induce lower NO release than EDTA-based irrigants.     

In vitro antibacterial activity of different endodontic irrigants

Abstract – Aim: The objective of this study was to compare in vitro the antibacterial activity of Tetraclean (mixture of doxycycline, citric acid and polypropylene glycol), Niclor 5 (5.25% sodium hypochlorite solution), Cloreximid (0.2% chlorhexidine and 0.2% cetrimide solution) and hydrogen peroxide 12 volumes on three endodontic pathogens associated with primary endodontic infections. Enterococcus faecalis, Streptococcus mutans and Staphylococcus aureus strains were selected to evaluate the antibacterial activity of endodontic irrigants by the agar disc‐diffusion test. Material and methods: Paper discs were saturated with each one of the test solutions (at room temperature and preheating at 50°C) and placed onto culture agar‐plates preadsorbed with bacterial cells and further incubated for 24 h at 37°C. The growth inhibition zones around each irrigant were recorded and compared for each bacterial strain. Kruskal–Wallis and Mann–Whitney tests were applied to compare the various groups. Results: At room temperature, Tetraclean showed significantly higher inhibition of bacteria growth than all other irrigants tested. Preheating at 50°C significantly increased growth inhibition for all the groups tested. At 50°C, hydrogen peroxide 12 volumes and Tetraclean showed significantly higher efficacy than all other irrigants tested. Conclusions: 50°C‐preheated hydrogen peroxide 12 volumes and Tetraclean showed highest inhibition of the bacterial growth.