Antibacterial substantivity of a new antibiotic‐based endodontic irrigation solution

The aim of this study was to evaluate the residual antibacterial activity of Tetraclean, MTAD and 5.25% sodium hypochlorite (NaOCl) in bovine root dentin in vitro. One hundred and ten dentin tubes prepared from bovine incisor teeth were infected in vitro for 14 days with Enterococcus faecalis. Thereafter, the specimens were divided into five groups as follows: Tetraclean; MTAD; 5.25% NaOCl; infected dentin tubes (positive control); and sterile dentin tubes (negative control). Dentin chips were collected using round burs into tryptic soy broth and after culturing, the number of colony‐forming units (CFU) was counted. The number of CFU in all experimental groups was minimum after treatment, and the results obtained were significantly different from each other at any time period (P < 0.05). The Tetraclean group showed the most effective antibacterial action at all five experimental periods (P < 0.05). MTAD group showed the least antibacterial activity after treatment. However, at days 7, 14, 21 and 28 MTAD showed more effective antibacterial action than NaOCl. In each group, the number of CFU increased significantly by time‐lapse (P < 0.05). In conclusion, the residual antibacterial activity of Tetraclean was significantly greater than MTAD and 5.25% NaOCl.     

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).     

In vitro antimicrobial activity of sodium hypochlorite and chlorhexidine against selected single-species biofilms

AIM: To investigate the antimicrobial activity of 2.5% and 5.25% sodium hypochlorite and 2.0% chlorhexidine gel and liquid as endodontic-irrigating substances against selected single-species biofilms. METHODS: Single-species biofilms of Enterococcus faecalis, Staphylococcus aureus, Candida albicans, Prevotella intermedia, Porphyromonas gingivalis, Porphyromonas endodontalis and Fusobacterium nucleatum were generated on a cellulose nitrate membrane placed on agar medium. The biofilms were then immersed in the endodontic-irrigating substances for 30 s and also for 5, 10, 15, 30 and 60 min, with and without mechanical agitation. Sterile saline was used as control. After each time period, the membrane filters were then transferred to tubes containing 2 mL of fresh broth medium plus neutralizers (in order to prevent the residual action of the tested substances). The micro-organisms were suspended using a vortex, and the inoculum was serially diluted 10-fold. Aliquots of the dilutions were plated on 5% sheep blood agar medium, and incubated under adequate gaseous conditions. Colony-forming units were calculated. The samples were compared using the Friedman and Tukey test, when necessary, at a significance level of P < 0.05. RESULTS: Mechanical agitation promoted the effectiveness of the antimicrobial agents, resulting in less time to eliminate the same micro-organisms, except for S. aureus with 2.5% NaOCl. Antimicrobial agents in liquid presentation, especially 5.25% NaOCl and 2% chlorhexidine, killed the tested micro-organisms more rapidly. Saline did not inhibit the growth of any of the tested micro-organisms, with or without agitation, being statistically different (P < 0.05) from NaOCl and chlorhexidine. P. intermedia, P. gingivalis, P. endodontalis and F. nucleatum were eliminated in 30 s by all antimicrobial agents, with our without agitation, in contrast with the facultative and aerobe strains. CONCLUSIONS: Mechanical agitation improved the antimicrobial properties of the chemical substances tested using a biofilm model, favouring the agents in liquid presentation, especially 5.25% NaOCl and 2% chlorhexidine.     

Comparison of the antimicrobial efficacy of 1.3% NaOCl/BioPure MTAD to 5.25% NaOCl/15% EDTA for root canal irrigation

The purpose of this study was to compare the antimicrobial efficacy of 1.3% NaOCl/BioPure MTAD to 5.25% NaOCl/15% EDTA for root canal irrigation. Twenty-six bilaterally matched pairs of human teeth were collected. The teeth were incubated with Enterococcus faecalis for 4 weeks. The teeth were divided into two experimental groups and one positive control group. The canals were instrumented and irrigated with either 5.25% NaOCl/15% EDTA or 1.3% NaOCl/BioPure MTAD. Bacterial samples were collected after instrumentation/irrigation and after additional canal enlargement. Statistical analysis of the data using the Wilcoxon Signed Rank test showed significant differences between the experimental groups. The first bacterial samples revealed growth in 0 of 20 samples with 5.25% NaOCl/15% EDTA irrigation and in 8 of 20 samples with 1.3% NaOCl/BioPure MTAD irrigation. Samples taken after additional canal enlargement revealed growth in 0 of 20 samples in 5.25% NaOCl/15% EDTA and in 10 of 20 samples in 1.3% NaOCl/BioPure MTAD group. This investigation showed consistent disinfection of infected root canals with 5.25% NaOCl/15% EDTA. The combination of 1.3% NaOCl/BioPure MTAD left nearly 50% of the canals contaminated with E. faecalis.     

An in vitro comparison of the antimicrobial effects of various endodontic medicaments on Enterococcus faecalis

The purpose of this in vitro study was to investigate the antimicrobial action of Dermacyn (Oculus Innovative Sciences, Petaluma, CA), BioPure MTAD (Dentsply Tulsa Dental, Johnson City, TN), 2% chlorhexidine (CHX; Ultradent, West Jordan, UT), and 5.25% sodium hypochlorite (NaOCl) against Enterococcus faecalis (American Type Culture Collection 4082). Eighteen Petri dishes of BHI agar were inoculated with E faecalis. Each Petri dish had five saturated paper disks placed. Four of the disks were saturated with a different test solution, and the last paper disk served as the control and was saturated with sterile distilled water. The plates were randomly distributed into two groups. Group one (n=9) was incubated aerobically and group 2 (n=9) was incubated anaerobically for 48 hours at 37 degrees C. The largest diameter of the zones of microbial inhibition was measured in millimeters and recorded. Statistical analysis was performed with repeated-measures analysis of variance. BioPure MTAD showed significantly (p<0.05) more zones of microbial inhibition than 5.25% NaOCl, 2% CHX, and Dermacyn. Sodium hypochlorite and CHX showed significantly (p<0.05) more zones of microbial inhibition than Dermacyn. The zone of inhibition between NaOCl and CHX was not significant (p>0.05). The control group showed no microbial inhibition.     

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 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.     

Surface tension comparison of four common root canal irrigants and two new irrigants containing antibiotic

The aim of this study is to compare the surface tension of four common endodontic irrigants: Moltendo EDTA 17%, Cetrexidin, Smear Clear, Sodium hypochlorite 5.25%, with the surface tension of MTAD and Tetraclean. Freshly produced MilliQ water was used as a reference. All measurements were performed following the Wilhelmy plate technique, using a Cahn DCA-322 Dynamic Contact Angle Analyzer at the temperature of 22 degrees C. MilliQ water, sodium hypochlorite 5.25%, and EDTA 17% had the highest surface tension, whereas those of Cetrexedin and Tetraclean has shown the lowest surface tension value. Both new irrigants, MTAD and Tetraclean, are capable of removing the smear layer. Thanks to their low surface tension, increasing the intimate contact of irrigant solutions with the dentinal walls, they may permit deeper penetration.     

Reduction in antimicrobial substantivity of MTAD after initial sodium hypochlorite irrigation

Potential intrinsic tetracycline staining of intraradicular dentin has been observed when BioPure MTAD was employed as the final irrigant after initial rinsing with NaOCl. This study examined the effect of NaOCl-MTAD interaction on the antimicrobial substantivity of MTAD in dentin. Dentin cores previously irrigated with either MTAD, or in conjunction with 1.3% NaOCl as an initial irrigant were placed on blood agar plates inoculated with Escherichia faecalis at 10(5) cfu/ml. Dentin cores irrigated with 1.3% NaOCl only, and autoclaved dentin disks were used as the respective positive and negative controls. After anaerobic incubation, the mean diameter of bacterial inhibition zones formed around the MTAD group was significantly larger than the NaOCl/MTAD group, which, in turn, was not significantly different from the NaOCl positive control. Oxidation of MTAD by NaOCl resulted in the partial loss of antimicrobial substantivity in a manner similar to the peroxidation of tetracycline by reactive oxygen species.     

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.     

The effect of passive ultrasonic activation of 2% chlorhexidine or 5.25% sodium hypochlorite irrigant on residual antimicrobial activity in root canals

Ninety-four single-canal roots were prepared using the step-down technique. Forty-two canals were irrigated with 2% chlorhexidine, 42 canals with 5.25% sodium hypochlorite (NaOCl), and 10 control canals with phosphate-buffered saline (PBS). The chlorhexidine and NaOCl groups were each then equally divided into a final irrigation group and a 1-min passive ultrasonic irrigation group. Canals were enlarged with a Parapost drill. The apical 3-5 mm was covered with nail polish. Canals were rinsed with PBS, dried, refilled with PBS, and stored. At 6 h, 20 microl of fluid was pipetted from each canal and placed into wells on agar plates, which were inoculated with Streptococcus sanguinis. The plates were incubated, and zones of inhibition were measured. Sampling was repeated at 24, 48, 72, 96, 120, 144, and 168 h. Residual antimicrobial activity with 2% chlorhexidine was statistically significantly superior to 5.25% NaOCl with irrigation alone and with final passive ultrasonic activation (p < 0.001). Chlorhexidine experimental groups demonstrated residual antimicrobial activity for as long as 168 h.     

The effect of exposure to irrigant solutions on apical dentin biofilms in vitro

This study assessed the effectiveness of different concentrations of sodium hypochlorite (NaOCl), 2% chlorhexidine (CHX) (Vista Dental Products, Racine, WI), and BioPure MTAD (Dentsply Endodontics-Tulsa Dental, Tulsa, OK). Intracanal contents were collected from 10 patients diagnosed with chronic apical periodontitis. The samples were cultured on hemisections of root apices to generate a polymicrobial biofilm. Each biofilm was separately immersed in 6% NaOCl, 3% NaOCl, 1% NaOCl, 2% CHX, 1% NaOCl followed by BioPure MTAD, and sterile phosphate buffered solution (PBS). SEM analysis showed 6% NaOCl and 3% NaOCl were capable of disrupting and removing the biofilm; 1% NaOCl and 1% NaOCl followed by MTAD were capable of disrupting the biofilm, but not eliminating bacteria; 2% CHX was not capable of disrupting the biofilm. Viable bacteria could not be cultured from specimens exposed to 6% NaOCl, 2 % CHX, or 1% NaOCl followed by BioPure MTAD. These results indicate that 6% NaOCl was the only irrigant capable of both rendering bacteria nonviable and physically removing the biofilm.     

In vitro antifungal efficacy of four irrigants as a final rinse

The purpose of this study was to compare the antifungal efficacy of 6% sodium hypochlorite (NaOCl), 2% chlorhexidine gluconate (CHX), 17% ethylene-diaminetetraacetic acid (EDTA), and BioPure MTAD as a final rinse on Candida albicans in vitro. Forty-eight, single rooted teeth were randomly divided into four groups each with a positive and a negative control. After root canal preparation, experimental teeth were inoculated with Candida albicans (ATCC 60193) and incubated for 72 hours. The groups were rinsed as follows: 1 ml of 6% NaOCl, 0.2 ml of 2% CHX, 1 ml of 17% EDTA, and 5 ml of BioPure MTAD. Aliquots from the experimental teeth were plated on Sabouraud 4% dextrose agar plates and colony-forming units were counted as a measure of antifungal activity. The results showed that 6% NaOCl and 2% CHX were equally effective and statistically significantly superior to BioPure MTAD and 17% EDTA (p < 0.05) in antifungal activity.     

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 temperature on sodium hypochlorite short-term stability, pulp dissolution capacity, and antimicrobial efficacy

The purpose of this study was to test some effects of preheating NaOCl solutions using a commercially available syringe heating device. Irrigating solution temperatures in 10-ml syringes were measured. Stability of 5.25, 2.62, and 1% NaOCl solutions for 60 min at 20, 45, and 60 degrees C was assessed using iodine/thiosulfate titration. Human pulp tissue dissolution capacity of a 1% NaOCl solution was gauged at the latter temperatures, and compared to corresponding values with a 5.25% solution at 20 degrees C. Killing efficacy of diluted NaOCl solutions against 48-h incubations of Enterococcus feacalis ATCC 29212 was compared at 45 degrees C and 20 degrees C. Using the heating device, a 20 degrees C solution reached 45 degrees C and 60 degrees C in 7 and 20 min, respectively. Solutions remained stable during the observation period. The 1% NaOCl solution at 45 degrees C dissolved pulp tissues as effectively as the 5.25% solution at 20 degrees C, while the 60 degrees C/1% solution was significantly more effective (p < 0.05). A 100-fold increase in killing efficacy was observed between corresponding NaOCl solutions at 20 degrees C and 45 degrees C.     

Comparative evaluation of endodontic irrigants against Enterococcus faecalis biofilms

The aim of this study was to compare the efficacy of root canal irrigants against E. faecalis biofilms using a novel in vitro testing system. Biofilms grown in a flow cell system were submerged in test irrigants for either 1 or 5 minutes. Statistical analysis revealed a significant relationship between test agent and percentage kill of the biofilm bacteria (P < 0.05). No statistically significant relationship between time and percentage kill was found. The percentage kill of the biofilm bacteria was: 6% NaOCl (>99.99%), 1% NaOCl (99.78%), Smear Clear (78.06%), 2% chlorhexidine (60.49%), REDTA (26.99%), and BioPure MTAD (16.08%). Post-hoc analysis showed a significant difference between 1% and 6% NaOCl, and all other agents including Smear Clear, 2% chlorhexidine, REDTA, and BioPure MTAD (P < 0.05). Within the parameters of this study, both 1% NaOCl and 6% NaOCl were more efficient in eliminating E. faecalis biofilm than the other solutions tested.     

Inhibitory activity of root canal irrigants against Candida albicans, Enterococcus faecalis and Staphylococcus aureus

The present study evaluated the antimicrobial activity of three root canal irrigants against Enterococcus faecalis, Candida albicans, and Staphylococcus aureus. These microorganisms were incubated in the presence of citric acid (6 and 10%), EDTA (17%), and NaOCl (0.5, 1.0, 2.5, and 5.25%). Agar diffusion tests were performed and redox indicator resazurin was used to evaluate the inhibitory effect of the irrigants on the metabolic activity of these microorganisms. The mean diameters of the inhibition zones for the C. albicans cultures were 11.6 mm (17% EDTA), 5.5 mm (0.5% NaOCl), 12.9 mm (1% NaOCl), 22.1 mm (2.5% NaOCl), and 28.5 mm (5.25% NaOCl). The mean diameters of the inhibition zones for E. faecalis were 2.8 mm (1% NaOCl), 5.4 mm (2.5% NaOCl), and 8.3 mm (5.25% NaOCl). For S. aureus, the mean values were 8.0 mm (17% EDTA), 3.0 mm (1% NaOCl), 8.8 mm (2.5% NaOCl), and 10.0 mm (5.25% NaOCl). Most of the irrigant solutions presented effective antimicrobial activity against C. albicans. A high inhibitory effect on the metabolic activity of E. faecalis was detected when the microorganisms were incubated with 17% EDTA. The same result was reached when S. aureus was incubated in the presence of > 2.5% NaOCl. Altogether, these results indicate that 2.5% and 5.25% NaOCl are microbicides against S. aureus while 0.5% and 1% NaOCl are only microbiostatic against the tested bacteria. The 6% and 10% citric acid as well as 17% EDTA did not affect the viability of any of the assayed microorganisms.     

Antimicrobial effects of various endodontic irrigants on selected microorganisms

Aim This study was undertaken to determine the antimicrobial effect of various endodontic irrigants against six selected microorganisms
Methodology Staphylococcus aureus , Enterococcus faecalis , Streptococcus salivarius , Str. pyogenes , Escherichia coli and Candida albicans were included in the study. Pre-sterilized Whatman paper discs, 6 mm in diameter and soaked with the test solution, were prepared and placed onto the previously seeded agar Petri plates. Each plate was incubated aerobically. A zone of inhibition was recorded for each plate and the results were analysed statistically.
Results 5.25% NaOCl was effective against all test microorganisms with a substantial zone of inhibition. Saline was always ineffective. Decreased concentration of NaOCl significantly reduced its antimicrobial effect. Cresophene showed a significantly larger ( P  > 0.05) average zone of inhibition compared to the other experimental irrigants. Alcohol had smaller but not significantly different zones of inhibition than chlorhexidine.
Conclusions 5.25% NaOCl was superior in its antimicrobial abilities compared with other irrigants used. A reduced concentration of NaOCl (0.5%) resulted in significantly decreased antimicrobial effects. When compared with 21% alcohol, 0.5% NaOCl and 2% chlorhexidine, paramonochlorophenol (cresophene) showed a greater antimicrobial effect.     

Comparison of 5.25% sodium hypochlorite, MTAD, and 2% chlorhexidine in the rapid disinfection of polycaprolactone-based root canal filling material

The purpose of this investigation was to compare the effectiveness of 5.25% sodium hypochlorite, MTAD, and 2% chlorhexidine in the rapid disinfection of Resilon pellets. Resilon pellets were randomly selected, contaminated with Enterococcus faecalis, and disinfected with 5.25% sodium hypochlorite, MTAD, or 2% chlorhexidine. Positive and negative controls were used. All samples were air dried, washed in sterile water, placed in centrifuge tubes containing BHI broth, and incubated at 37 degrees C for up to 7 days. Broths were visually checked for turbidity and scored for growth. Samples from each experimental and control group were randomly chosen, plated, incubated at 37 degrees C, and checked visually for growth. If growth occurred, a gram stain was performed to identify bacterial species. Results indicate that 5.25% sodium hypochlorite, MTAD, and 2% chlorhexidine were all effective in the rapid disinfection of Resilon and gutta-percha pellets, and a 1-minute immersion was sufficient to disinfect.