Matilde Ruiz-Linares

Antimicrobial Activity of a Sodium Hypochlorite/Etidronic Acid Irrigant Solution

INTRODUCTION The aim of this study was to evaluate the antimicrobial activity of a 2.5% sodium hypochlorite (NaOCl)/9% etidronic acid (HEBP) irrigant solution on Enterococcus faecalis growing in biofilms and a dentinal tubule infection model. METHODS The antimicrobial activity of the solutions 2.5% NaOCl and 9% HEBP alone and associated was evaluated on E. faecalis biofilms grown in the Calgary biofilm model (minimum biofilm eradication concentration high-throughput device). For the dentinal tubule infection test, the percentage of dead cells in E. faecalis-infected dentinal tubules treated with the solutions for 10 minutes was measured using confocal laser scanning microscopy and the live/dead technique. Available chlorine and pH of the solutions were also measured. Distilled water was used as the control. Nonparametric tests were used to determine statistical differences. RESULTS The highest viability was found in the distilled water group and the lowest in the NaOCl-treated dentin (P < .05). Both NaOCl solutions killed 100% of the E. faecalis biofilms and showed the highest antimicrobial activity inside dentinal tubules, without statistical differences between the 2 (P < .05). The HEBP isolated solution killed bacteria inside dentinal tubules but did not present any significant effect against E. faecalis biofilms. The incorporation of HEBP to NaOCl did not cause any loss of available chlorine within 60 minutes. CONCLUSIONS HEBP did not interfere with the ability of NaOCl to kill E. faecalis grown in biofilms and inside dentinal tubules.

Antibacterial and Anti-biofilm Activity of AH Plus with Chlorhexidine and Cetrimide

INTRODUCTION The use of root canal filling materials with antibacterial activity can be considered beneficial to reduce the remaining microorganisms in the root canal system, where Enterococcus faecalis is often found, and prevent recurrent infection. The aim of this study was to evaluate the antimicrobial activity and capacity for inhibiting E. faecalis biofilm formation of AH Plus, alone and mixed with chlorhexidine (CHX), cetrimide (CTR), and combinations of the two. METHODS AH Plus alone and mixed with 1% and 2% CHX, 0.1%-0.5% CTR, and combinations of both were tested to assess antimicrobial activity by a modified direct contact test and determine inhibition of E. faecalis biofilm formation at 24 hours. The results were expressed as log10 viable counts. Eradication and inhibition of biofilm formation were understood as no bacterial growth or log10 reduction = 5 with respect to the control (AH Plus alone). RESULTS AH Plus + CHX showed a low antimicrobial activity with respect to the control (at 2%, log10 reduction = 1.30). None of the tested concentrations achieved eradication or inhibition of biofilm. AH Plus + CTR showed a direct relationship of concentration-antimicrobial effect, reaching a log10 reduction of 2.92 at 0.5% and inhibition of biofilm formation at 0.2%. With the combination CHX + CTR, lower concentrations were needed for the same effect, and eradication and inhibition of biofilm were achieved. CONCLUSIONS The addition of CHX, CTR, or some combination of both to AH Plus confers it with bactericidal and anti-biofilm activity against E. faecalis.

Reduction in Enteroccocus faecalis counts – a comparison between rotary and reciprocating systems

AIM To compare the chemomechanical reduction and regrowth of Enterococcus faecalis between rotary and reciprocating systems in root canal preparation. METHODOLOGY Seventy-six single-rooted human mandibular premolars were selected and standardized to 15 mm in length. Root canals were enlarged up to a size 25 K-file and irrigated with distilled water and then were infected with E. faecalis for 4 weeks. The specimens were divided into 3 groups (n = 24) for instrumentation with Mtwo, Twisted File and WaveOne. Each group was further divided into two subgroups (n = 12) according to the irrigant used: distilled water or 5.25% sodium hypochlorite (NaOCl). Before and after rotary preparation, microbiological samples were collected using three sterilized paper points, and efficacy was expressed as reduction in percentage. The proportion of grown samples for 60 days was evaluated using nonparametric Kaplan-Meier survival analysis. Differences amongst groups were tested using the log-rank test at a significance level of 0.05. RESULTS In the main root canal, the percentage reduction in the distilled water and 5.25% NaOCl groups ranged from 95.9% to 100%, with no significant differences amongst the three systems (P > 0.05). The bacterial regrowth in NaOCl groups revealed that Mtwo had the lowest number of samples regrown at 60 days, giving statistically significant differences with respect to Twisted File (P = 0.029) and WaveOne (P = 0.005). CONCLUSIONS Reciprocating and rotary systems resulted in similar percentage reduction in E. faecalis when using either distilled water or 5.25% NaOCl solution. Over time, the Mtwo system was more effective regarding disinfection.

Antimicrobial residual effects of irrigation regimens with maleic acid in infected root canals

BackgroundThe success of endodontic treatment depends largely on the control of microorganisms present in infected root canals. The aim of this study was to determine the residual antimicrobial activity of several final irrigation protocols with 7% maleic acid (MA) alone and combined with chlorhexidine (CHX), cetrimide (CTR) or both, in root canals infected with Enterococcus faecalis. Biofilms of E. faecalis were grown in uniradicular roots for 4 weeks. A total of 72 specimens were divided into 5 experimental groups according to the final irrigation regime used: Group 1: 2.5% NaOCl; Group 2: 7% MA; Group 3: 7% MA + 0.2% CTR; Group 4: 7% MA + 2% CHX; Group 5: 7% MA + 0.2% CTR + 2% CHX; and Control group: 0.9% saline solution. Twelve roots (2/group) that were instrumented and not infected served as negative or sterility controls. The proportion of ungrown samples over 60 days was evaluated using non-parametric Kaplan-Meier survival analysis. Differences among groups were tested using the log-rank test (p< 0.05).ResultsThe association of MA with CHX and CHX + CTR as final irrigating solutions showed the best results, 5 and 4 samples out of 12, respectively, and without differences between them (p = 0.928), followed by 7% MA + 0.2% CTR with 7 out of 12. The 7% MA (11/12) group showed significant differences with respect to the groups in which MA was combined with CHX (p < 0.005) and CHX + CTR (p < 0.005).ConclusionFinal irrigating solutions of 7% MA combined with 2% CHX or 2% CHX + 0.2% CTR were found to effectively improve antimicrobial root canal disinfection.

The effect of benzalkonium chloride additions to AH Plus sealer. Antimicrobial, physical and chemical properties

OBJECTIVES The aim of this study was to determine the antimicrobial and antibiofilm activities and physicochemical properties of AH Plus sealer mixed with different concentrations of benzalkonium chloride (BC). METHODS AH Plus was tested alone and mixed with 1%, 2% and 3% of BC. The antimicrobial and antibiofilm activities of the sealers against Enterococcus faecalis were evaluated by the direct contact test (DCT) and by confocal laser scanning microscopy, respectively. Setting time, flow and solubility were assessed according to ANSI/ADA specifications. Microhardness and contact angle tests were also performed. The chemical changes of the sealers were evaluated by X-ray diffraction analysis, and both Fourier transform infrared spectroscopy (FT-IR) and attenuated total reflectance Fourier transform infrared (ATR FT-IR). RESULTS AH Plus+3% BC was the only sealer to promote total elimination of E. faecalis and the biovolume in this group was significantly lower than in the rest of the sealers (p>0.05). The physical properties of the sealers were according to the ANSI/ADA specifications. The microhardness decreased significantly when BC was added and a significant reduction in contact angle was obtained when incorporating 2% and 3% BC (p<0.05). No phase changes were observed with the modified sealers. CONCLUSIONS The addition of 2% or higher concentrations BC to AH Plus showed antimicrobial and antibiofilm activities without affecting the properties specified in ANSI/ADA standards. However, additives to the root canal sealer altered other physical and chemical properties that are not commonly found in the literature to evaluate filling materials. CLINICAL SIGNIFICANCE The present study highlights that the antimicrobial properties of AH Plus can be significantly improved with the addition of BC. Testing beyond what is specified in standards may be indicated.

Antimicrobial activity of alexidine, chlorhexidine and cetrimide against Streptococcus mutans biofilm.

BackgroundThe use of antimicrobial solutions has been recommended to disinfect demineralized dentin prior to placing the filling material. The aim of this study was to evaluate the ability of several antimicrobials in controlling Streptococcus mutans (SM) biofilm formed in dentin.MethodsAntimicrobial activity of 0.2% and 2% chlorhexidine (CHX), 0.2% cetrimide (CTR) and 0.2%, 0.5%, 1% and 2% alexidine (ALX) was assayed on 1-week SM biofilm formed on standardized coronal dentin blocks. Results of SM biofilm antimicrobial activity by different protocols were expressed as the kill percentage of biofilm and the term “eradication” was used to denote the kill of 100% of the bacterial population. To compare the efficacies of the different protocols the Student t test was used, previously subjecting data to the Anscombe transformation.ResultsAll ALX concentrations tested and 0.2% CTR achieved a kill percentage higher than 99%, followed by 2% CHX with percentages above 96% (no statistically significant difference among them). Whereas 2% ALX and 0.2% CTR respectively showed eradication in 10 and 9 of the twelve specimens, 0.2% CHX did not produce eradication in any case.ConclusionsThe present study shows that, when used for one minute, 2% and 1% alexidine, and 0.2% cetrimide, achieve eradication of Streptococcus mutans biofilm in most specimens when applied to a dentin-volumetric model.

Antimicrobial Substantivity of Alexidine and Chlorhexidine in Dentin

INTRODUCTION The use of irrigating solutions with long-lived activity helps to avoid failure of endodontic therapy. The aim of the present study was to evaluate and compare the antimicrobial substantivity against Enterococcus faecalis by using as carrier a dentin-volumetric unit exposed to chlorhexidine (CHX) and alexidine (ALX). METHODS Standardized dentin blocks of human molars were treated for 1 minute with 0.5% CHX, 2% CHX, 1% ALX, and 2% ALX. They were exposed to E. faecalis in suspension to determine antimicrobial substantivity during a period of 80 days. Results were analyzed by means of Kaplan-Meier survival analysis (P < .05). RESULTS ALX obtained the best results, with differences that were significantly different from CHX regardless of the concentration. Although all the samples treated with CHX were contaminated at the end of the study period, a direct relationship was seen between concentration and survival time. There was no statistically significant difference between 2% and 1% ALX. CONCLUSIONS The present study shows that 2% and 1% ALX used for 1 minute provide longer antimicrobial substantivity against E. faecalis than CHX when applied to 2% and 0.5%.

Physical Properties of AH Plus with Chlorhexidine and Cetrimide

INTRODUCTION The use of root canal filling materials with antibacterial activity could be considered beneficial to reduce any remaining microorganisms and prevent recurrent infection. The aim of this study was to evaluate the physical properties of AH Plus (Dentsply DeTrey, Konstanz, Germany) alone and mixed with 1% or 2% chlorhexidine (CHX); 0.1%, 0.2%, 0.3%, and 0.5% of cetrimide (CTR); and combinations of both. METHODS Setting time, flow, solubility, and radiopacity of AH Plus and modified AH Plus were evaluated following the American National Standards Institute/American Dental Associations requirements (ANSI/ADA Specification No. 57/2000). Five samples of each material were tested for each property. The hypothesis of equality among groups was rejected by an analysis of variance test, and then a post hoc Tukey test was performed. RESULTS AH Plus and modified AH Plus gave values that are within the required standards. Overall, results indicate that CTR increased setting time, but CHX alone and CHX + CTR reduced it. CHX increased the flow, whereas CTR reduced it, and their combinations gave intermediate values comparable with those of AH Plus. Concerning solubility, the weight loss in all samples was under the 3% limit set by the ANSI/ADA. In comparison with AH Plus alone, radiopacity gave slightly lower values with CHX, higher values with CTR, and similar values with combinations. CONCLUSIONS The addition of CHX, CTR, and combinations of both to AH Plus did not alter the physical properties specified by ANSI/ADA requirements.

Bacterial leakage in root canals filled with AH Plus and dentine bonding agents

Abstract Objective. The aim of this study was to compare the efficacy of different dentine adhesives in delaying the coronal bacterial leakage of Enterococcus faecalis in filled root canals. Materials and methods. Ninety-five lower incisors of patients >65 years of age were instrumented using the ProTaper® system and were irrigated with 1 mL of 2.5% sodium hypochlorite (NaOCl) alternated with 1 mL 17% EDTA between each file change. Final irrigation was performed with 5 mL of 17% EDTA and then flushed with 5 mL of distilled water. The teeth were randomly divided into five experimental groups (n = 15/group) and one of the following dentine adhesives was applied: (1) AdheSE®; (2) Excite® DSC; (3) Clearfil™ Protect Bond; (4) One Coat 7.0; or (5) Control group without adhesive. After filling the root canals, the samples were mounted on a double chamber device to evaluate the bacterial filtration of E. faecalis during a period of 240 days. The results underwent non-parametric Kaplan-Meier survival analysis and comparisons among groups were done using the Log-Rank test. Results. At 240 days, E. faecalis was detected in samples of all groups in the lower chamber. The highest survival value was obtained by One Coat 7.0, giving statistically significant differences from the other groups, whereas Clearfil™ Protect Bond, AdheSE® and Excite® DSC showed similar behaviours, likewise similar to the Control group. Conclusions. One Coat 7.0 adhesive system provides the longest survival value to delay E. faecalis coronal leakage in filled root canals.

Antibiofilm Activity of Sodium Hypochlorite and Alkaline Tetrasodium EDTA Solutions

Introduction The aim of this study was to determine the antimicrobial activity of mixed alkaline tetrasodium EDTA (EDTANa4)/sodium hypochlorite (NaOCl) solutions with and without the addition of cetrimide (CTR) against Enterococcus faecalis biofilms. Methods The antimicrobial solutions were evaluated on a 3‐week biofilm of E. faecalis grown on radicular dentin blocks. The irrigating solutions were 2.5% NaOCl group, 20% EDTANa4 group, 10% EDTANa4 group, 2.5% NaOCl/10% EDTANa4 group, 2.5% NaOCl/5% EDTANa4 group, 2.5% NaOCl/10% EDTANa4/0.2% CTR group, 2.5% NaOCl/5% EDTANa4/0.2% CTR group, and 0.9% saline solution group. Cell viability was determined by adenosine triphosphate assay, and culture techniques were used to determine colony‐forming unit (CFU) counts. In groups with positive growth, the Pearson linear correlation coefficient r (−1 ≤ r ≤ 1) between relative light units (RLUs) and log10 (CFUs + 1) was calculated. The results of the bioluminescence adenosine triphosphate assay and CFUs of the groups were analyzed by means of analysis of variance and the Duncan test. Results There was Pearson correlation between RLUs and log10 (CFUs + 1). A similar reduction of RLUs in all NaOCl groups was seen, giving statistical differences with respect to the control. In the NaOCl groups there was no bacterial growth. The lowest antimicrobial efficacy was found for the EDTANa4 groups. Conclusions Mixed alkaline EDTANa4/NaOCl solutions with and without the addition of CTR do not interfere with the antimicrobial activity of NaOCl. HIGHLIGHTSThe ATP assay correlates well with the microbial counts to determine E. faecalis on root dentin.EDTANa4 with NaOCl and NaOCl + CTR effectively control biofilms of E. faecalis on dentin.Such solutions could constitute an effective alternative for use as a single irrigating solution.