Efficacy of chlorhexidine- and calcium hydroxide-containing medicaments against Enterococcus faecalis in vitro

OBJECTIVE: We sought to assess the efficacy of chlorhexidine (CHX) and calcium hydroxide, Ca(OH)(2), against Enterococcus faecalis in vitro. STUDY DESIGN: The effect of CHX (0.2% and 2% in gel or solution) and Ca(OH)(2) (alone or with 0.2% CHX gel) was evaluated by using the agar diffusion test and an in vitro human root inoculation method, to measure zone of inhibition or bacterial growth with optical density analysis, respectively. For optical density analysis, samples from infected root canals were collected after 7 days of medication and were cultured for 24 hours in brain-heart infusion to detect viable bacteria. RESULTS: In the agar diffusion test, CHX was effective against E faecalis in a concentration-dependent fashion, but Ca(OH)(2) alone had no effect. In the root canal inoculation test, CHX was significantly more effective against E faecalis than Ca(OH)(2) was (P < .05), but there were no significant differences between the modes of medication or concentrations of CHX. CONCLUSIONS: CHX is effective against E faecalis in vitro. Further in vivo studies are needed to confirm the value of CHX in clinical treatment.     

Antimicrobial substantivity of bovine root dentin exposed to different chlorhexidine delivery vehicles

Root canal dentin acquires antimicrobial substantivity after exposure to chlorhexidine gluconate (CHX) for 1 wk. Therefore development of a vehicle for delivery of CHX as an intracanal medication is desirable. This in vitro study assessed the efficacy of two CHX delivery vehicles, a controlled-release device and a gel, to affect antimicrobial substantivity of bovine root dentin. Sixty bovine incisor root specimens were prepared with standardized length (10 mm) and canal diameter (3.3 mm), and coated externally with nail polish. Specimens were divided into four equal groups and their canals medicated for 7 days with either: (i) an experimental controlled-release device containing 25% CHX that was immersed in sterile saline; (ii) 2% CHX gel; or (iii) Ca(OH)2 paste. Sterile saline was used as the positive control. After medication, the canals of the specimens were inoculated with Enterococcus faecalis for 21 days. Root canal dentin samples ranging in depth from 0.1 to 0.45 mm were then obtained using sterile round burs of ascending diameter. Each dentin sample was placed in a separate test tube containing Brain Heart Infusion broth and incubated for 24 h. The optical density (OD) of the broth was then measured spectrophotometrically at 540 nm. The positive control showed significantly higher mean OD values (one-way ANOVA and Tukey's Studentized Range Test; p < 0.001) than the three test groups. The CHX controlled-release device group showed significantly lower OD values than the Ca(OH)2 group; however only at dentin depths up to 0.2 mm. In contrast, the CHX gel group consistently showed significantly lower OD values than both the CHX controlled-release device and Ca(OH)2 groups. These results suggest that bovine root canals medicated with 2% CHX gel for 7 days acquire antimicrobial properties for at least 21 days.     

Molecular- and culture-based comparison of the effects of antimicrobial agents on bacterial survival in infected dentinal tubules

The aim of this study was to evaluate the effect of root canal obturation with or without prior calcium hydroxide (Ca(OH)(2)) or 2% chlorhexidine (CHX) on the persistence of bacteria or its DNA in infected dentinal tubules. Canals of 85 extracted teeth were instrumented and inoculated with 10(4) cells/mL of Enterococcus faecalis. Teeth were incubated at 37 degrees C for 21 days and divided into 3 groups of 25 teeth plus controls. Teeth in group 1 were obturated immediately with gutta-percha (GP) and AH-Plus (Maillefer, Dentsply, Tulsa, OK). In group 2, Ca(OH)(2) was placed for 7 days before obturation. In group 3, 10 minutes of irrigation was performed with CHX performed before obturation. After incubation, GP was removed, and dentin specimens were collected and analyzed with culturing and polymerase chain reaction (PCR). No growth occurred in any cultures. By using PCR, E faecalis was detected in fewer roots in group 3 than in groups 1 or 2 (chi(2), p = 0.05); 2% CHX treatment followed by obturation was more effective in removing E faecalis DNA than placement of Ca(OH)(2) or immediate obturation.     

Control of microorganisms in vitro by endodontic irrigants

The aim of this study was to determine the minimum inhibitory concentration (MIC) and antimicrobial effectiveness by the direct exposure test of 4 endodontic irrigants [1% sodium hypochlorite (NaOCl), 2% chlorhexidine (CHX), 1% calcium hydroxide (Ca(OH)2; prepared with 1 g of Ca(OH)2 and 100 mL of sterile distilled water), a solution of Ca(OH)2 + detergent (HCT20)] for S. aureus, E. faecalis, P. aeruginosa, B. subtilis, C. albicans and a mixed culture. Microbial growth was analyzed by two methods: turbidity of the culture medium that was confirmed by Gram stain and subculture in a specific nutrient broth. In the dilution test, NaOCl solution showed MIC equal to 0.1% for S. aureus, E. faecalis, P. aeruginosa and C. albicans and equal to 1% for B. subtilis and the mixed culture. CHX (2%) presented MIC equal to 0.000002% for S. aureus, 0.02% for E. faecalis, B. subtilis, C. albicans and the mixed culture and 0.002% for P. aeruginosa. Ca(OH)2 solution (1%) showed MIC greater than 1% for all the microorganisms except P. aeruginosa for which it was equal to 1%. Calcium hydroxide solution + detergent showed MIC equal to 4.5 mL for S. aureus, P. aeruginosa, B. subtilis, C. albicans and the mixed culture and greater than 4.5 mL for E. faecalis. In the direct exposure test, NaOCl had better antimicrobial effectiveness for all microorganisms at all times. CHX (2%) was effective for S. aureus, E. faecalis and C. albicans at all times, and ineffective for P. aeruginosa, B. subtilis and the mixed culture. The other solutions showed the worst results.     

In vitro evaluation of the antimicrobial activity of calcium hydroxide combined with chlorhexidine gel used as intracanal medicament

The aim of this study was to investigate the antimicrobial activity of calcium hydroxide (Ca(OH)2) combined with 2% chlorhexidine gluconate (CHX) gel against endodontic pathogens and to compare the results with the ones achieved by Ca(OH)2 mixed with sterile water and by CHX gel alone. Two methods were used: the agar diffusion test and the direct contact test. Ca(OH)2 + 2% CHX gel produced inhibitory zones ranging from 2.84 to 6.5 mm, and required from 30 seconds to 6 hours to eliminate all tested microorganisms. However, 2% CHX gel showed the largest microbial growth zones from 4.33 to 21.67 mm, and required 1 minute or less to inhibit all tested microorganisms. A paste of Ca(OH)2 plus sterile water inhibited only the microorganisms with which it was in direct contact and required from 30 seconds to 24 hours to kill all tested microorganisms. In conclusion, 2% CHX gel + Ca(OH)2 showed better antimicrobial activity than Ca(OH)2 manipulated with sterile water.     

Antimicrobial efficacy of chlorhexidine and two calcium hydroxide formulations against Enterococcus faecalis

The purpose of this study was to investigate the efficacy of chlorhexidine (CHX) and calcium hydroxide (Ca(OH2) against Enterococcus faecalis in vitro. Extracted single-rooted human teeth were instrumented up to size 40. After removal of the smear layer, an inoculum of E. faecalis was inserted into the root canals. After incubation, the inoculum was removed and the root canals were filled with one of three different disinfectants: Ca(OH2 paste, CHX 2%, and a mixture of CHX and Ca(OH2 paste (n = 10 in each group). Control teeth were filled with water of standardized hardness (n = 10). The teeth were then incubated for 3 days. After incubation, each root canal was instrumented, and the removed dentin was examined microbiologically. CHX was significantly more effective against E. faecalis than was Ca(OH2 paste or a mixture of CHX with Ca(OH2 paste (p < 0.05). There was no increase in the efficiency of Ca(OH2 paste when CHX was added (p > 0.05). The results suggest that CHX is effective in the elimination of E. faecalis from dentinal tubules under the conditions of this study.     

Antimicrobial effect and pH of chlorhexidine gel and calcium hydroxide alone and associated with other materials

The purposes of this study were to evaluate the effectiveness of 2% chlorhexidine (CHX) gluconate gel, calcium hydroxide [Ca(OH)2] and their combination with iodoform and zinc oxide powder as intracanal medications against select microorganisms, and to measure the pH changes caused by these medications. Antimicrobial activity was determined by the agar diffusion method. The zones of growth inhibition were measured and the results were analyzed statistically by Kruskal-Wallis test (p<0.05). The pH of the pastes was measured right after preparation, after 24 h and 1 week later. The largest mean zones of microbial inhibition were produced by 2% CHX gel, followed by Ca(OH)2 + 2% CHX gel + iodoform, Ca(OH)2 + 2% CHX gel, Ca(OH)2 + 2% CHX gel + zinc oxide, and Ca(OH)2 + water. The mean pH of all medications stayed above 12.0 during the whole experiment, except for CHX gel (pH=7.0). The results of this study showed that all medications had antimicrobial activity, but the most effective against the tested microorganisms were 2% CHX gel, followed by its combination with Ca(OH)2 and iodoform.     

Comparative analysis of endodontic pathogens using checkerboard hybridization in relation to culture

Background/Aim:  The purpose of this study was to detect bacterial species and to quantify the total number of bacteria from samples of infected root canals before (S1) and after chemo-mechanical preparation using 2% chlorhexidine (CHX) gel as auxiliary chemical substance (S2) and after 7 days of intracanal dressing (S3) to compare microbial changes.
Method:  Twenty-four teeth were selected for this study. Chemo-mechanical preparation was performed using 2% CHX gel, then three different intracanal medicaments [M1: Ca(OH)₂ paste; M2: 2% CHX gel; and M3: Ca(OH)₂ paste plus 2% CHX gel] were used for 7 days. Checkerboard DNA–DNA hybridization was performed to detect 40 bacterial species. Aerobic and anaerobic culture techniques were used to determine the bacterial community by counting the colony-forming units (CFU).
Results:  The species most frequently identified by checkerboard in S1 were: Fusobacterium nucleatum ssp . polymorphum , Treponema socranskii ssp. socranskii , Parvimonas micra and Enterococcus faecalis. In S2 and S3 a total of eight different species were identified; and only one of them was gram-positive ( E. faecalis ). Microorganisms were not identified after use of M2 for 7 days. The quantification obtained on agar plates ranged from 4 × 10⁵ to 2.6 × 10⁶ CFU/ml in S1, mean CFU was reduced by 99.96% in S2, and there was no statistical difference between the CFU in S2 and S3.
Conclusion:  The antibacterial effect of the mechanical preparation supplemented by the use of an antibacterial auxiliary substance greatly reduced the microorganisms in the main root canal.     

In vitro evaluation of the effectiveness of irrigants and intracanal medicaments on microorganisms within root canals

AIM: To evaluate in vitro the effectiveness of sodium hypochlorite (NaOCl), chlorhexidine (CHX) and five intracanal medicaments on microorganisms within root canals. METHODOLOGY: Ninety-six human single-rooted extracted teeth were used. After removing the crowns, canal preparation was completed and the external root surfaces were coated with epoxy resin. Following sterilization, the teeth were contaminated with Candida albicans and Enterococcus faecalis, and were incubated at 37 +/- 1 degrees C for 7 days. The teeth were divided according to the irrigant solution or intracanal medicament: group 1, sterile physiologic solution (SPS) and calcium hydroxide (Ca(OH)2) paste; group 2, SPS and camphorated paramonochlorophenol (CPMC); group 3, SPS and tricresol formalin; group 4, SPS and CaOH2 + CPMC paste; group 5, SPS and PMC furacin; group 6, 2.5% NaOCl without intracanal medication; group 7, 2.0% CHX without intracanal medication and group 8, SPS without intracanal medication (control group). Microbiological samples were collected with sterile paper points, and bacterial growth was determined. The data were submitted to the analysis of variance (anova, P = 0.05). RESULTS: For C. albicans, groups 3 and 8 were statistically less effective than groups 1, 2, 4 and 5 (Kruskal-Wallis (K-W) = 65.241; gl = 7; P = 0.001). For E. faecalis, groups 6 and 8 were statistically less effective than groups 1-4 and 7 (K-W = 61.048; gl = 7; P = 0.001). CONCLUSIONS: Ca(OH)2 + CPMC paste was the most effective intracanal medicament for the elimination of the two microorganisms; 2.0% CHX solution was more effective than 2.5% NaOCl against E. faecalis.     

Antibacterial efficacy of calcium hydroxide and chlorhexidine gluconate irrigants at 37 degrees C and 46 degrees C

This study investigated the ability of two endodontic irrigants to eliminate Enterococcus faecalis from dentinal tubules, and whether their antimicrobial action was enhanced by heat. The lumens of disks prepared from extracted bovine roots were infected with E. faecalis and incubated for 72 h. Specimens were then filled with saline, 10% calcium hydroxide (Ca(OH)2), or 0.12% chlorhexidine gluconate (CHX) at 24 degrees C or 46 degrees C and incubated at 37 degrees C or 46 degrees C. The samples were then pulverized and plated to quantify residual bacteria. No statistical difference (p > 0.05) in bacterial growth was seen between the two saline groups, or between the two medication groups at a given temperature. CHX and Ca(OH)2 at either temperature produced significantly less growth than either saline group, and CHX or Ca(OH)2 at 46 degrees C produced significantly less growth than either group at 37 degrees C. Heat enhanced the antibacterial action of both experimental irrigants against E. faecalis, but heating saline produced no increase in bactericidal effect.     

Bacterial leakage in roots filled with different medicaments and sealed with Cavit

The aim of this study was to evaluate the time required by four different root canal medications coupled with the temporary filling material Cavit (ESPE, Seefeld, Germany) to prevent penetration of bacteria into the root canal. There were 145 roots prepared in a standardized manner. Four groups with 15 samples each were dressed with calcium hydroxide (Ca(OH)(2)), a 5% chlorhexidine gel (CHX), a chloromono-campherphenolic compound (ChKM), and Ledermix (LM), respectively, and sealed with Cavit. Four control groups contained identical medications but the roots were left unsealed. The 25 remaining roots served as additional controls. A standard setup for bacterial leakage studies was chosen with Staphylococcus epidermidis as test strain. Cavit application resulted in a significantly better seal compared with the unsealed groups. In the Cavit-sealed groups, all groups differed significantly from one another except for the CHX and the ChKM groups. The Ca(OH)(2) medicated roots provided the longest protection (median of 36 days), followed by the Ledermix-group (27 days) and the CHX (18 days) or ChKM groups (19 days). It may be concluded that Cavit-sealed and medicated root canals do not provide adequate protection against bacterial leakage for more than 1 month.     

The effect of antibiotics and endodontic antimicrobials on the polymerase chain reaction

The effectiveness of endodontic antimicrobial treatment could be determined using sensitive molecular methods. The purpose of this study was to determine if antibiotics or endodontic reagents interfere with the ability of PCR to detect Enterococcus faecalis in vitro. Amoxicillin (25 mg/ml), clindamycin (15 mg/ml), tetracycline (25 mg/ml), doxycycline (10 mg/ml), calcium hydroxide, 1% buffered sodium hypochlorite (NaOCl1), 3% and 6% unbuffered NaOCl (NaOCl3 and NaOCl6), 2% chlorhexidine (CHX), 5% tincture iodine (TI), 2% iodine potassium iodide (IKI), chloroform (CF), 70% ethyl alcohol, 5% sodium thiosulphate, 5% citric acid or saline were added to 10 or 10 cells/ml E. faecalis ATCC 19433 for 1 h (1 wk for Ca(OH)2). Using PCR, all specimens were positive except for NaOCl3 and NaOCl6. PCR with Ca(OH)2 was positive with 10 cells/ml but negative with 10 cells/ml. The following reagents yielded negative culturing results: all antibiotics, Ca(OH)2, CHX, IKI, TI, NaOCl3, NaOCl6, and CF. BacLight nuclear staining revealed the presence of viable cells in all PCR positive, culture negative combinations, except for those with CF. Therefore, in the presence of threshold values of bacterial concentrations, all reagents tested except for NaOCl3 and NaOCl6 do not interfere with the detection of E. faecalis using PCR.     

Tissue-dissolution capacity and dentin-disinfecting potential of calcium hydroxide mixed with irrigating solutions

OBJECTIVE: The goal of this study was to compare the tissue-dissolution potential and antibacterial effectiveness of a conventional Ca(OH)(2)/saline paste with equivalent Ca(OH)(2)/NaOCl and Ca(OH)(2)/chlorhexidine digluconate medications. STUDY DESIGN: Tissue specimens were obtained from freshly dissected pig palates. Tissue pieces of similar form and weight were incubated in air-tight containers with Ca(OH)(2) pastes or solutions proper for up to 7 days. Antimicrobial testing was performed in dentin blocks infected with Enterococcus faecalis. Medicated, sealed dentin specimens were incubated for 1 and 5 days, and bacterial growth was tested at different dentin depths. RESULTS: Up to day 4, the Ca(OH)(2)/irrigating solution mixtures dissolved tissue more effectively than the conventional Ca(OH)(2)/saline paste. After 7 days, however, no statistically significant differences were found between the saline and hypochlorite mixtures, but the Ca(OH)(2)/chlorhexidine medication was significantly less effective. Dentin block disinfection was quicker and more thorough with the Ca(OH)(2)/chlorhexidine or the Ca(OH)(2)/NaOCl than with the Ca(OH)(2)/saline paste. CONCLUSION: Ca(OH)(2)/irrigant mixtures under investigation appear more advantageous than the conventional Ca(OH)(2)/saline mixture, and merit further investigation in a clinical study.     

Removal efficacy of various calcium hydroxide/chlorhexidine medicaments from the root canal

Aim To compare the efficiency of removing calcium hydroxide [Ca(OH)₂]/chlorhexidine (CHX) (gel), Ca(OH)₂/CHX (solution) and Ca(OH)₂/saline pastes with the use of instrumentation and irrigation with sodium hypochlorite and ethylene diamine tetraacetic acid (EDTA) solutions. Moreover the role of the patency file in the cleanliness of the apical third of the root canal was evaluated. Methodology Sixty‐four human single‐rooted teeth with straight canals were used. Root canal preparation was performed with a stepback technique using Hedström (H) files. Teeth were randomly assigned to three groups and subsequently filled with one of the pastes: Ca(OH)₂/CHX (gel), Ca(OH)₂/CHX (solution) and Ca(OH)₂/saline paste. The medicaments were removed 10 days later using instrumentation and irrigation with 1% sodium hypochlorite and 17% EDTA, with or without obtaining patency of the apical foramen with a size 10 H‐file. The crowns were removed at the cemento‐enamel junction and the roots were grooved longitudinally and split into halves. Images of all halves were acquired with the use of a flatbed scanner. A scoring system of 1 to 4 was used to assess the amount of residue on the cervical, middle and apical third of the canal. Data were subjected to statistical analysis using Kruskal–Wallis and Mann–Whitney tests, with Bonferroni correction, at 95% confidence level (P < 0.05). Results Remnants of medicament were found in all experimental teeth regardless of the experimental material used and the use of the patency file. When examining the root canal as a whole, Ca(OH)₂/CHX (gel) paste was associated with significantly larger amount of residue, whereas the Ca(OH)₂/CHX (solution) paste was associated with less amount (P < 0.05) than the other two medicaments with or without the use of a patency file. Conclusions None of the techniques used in this study removed the inter‐appointment root canal medicaments effectively; the use of the patency file facilitated removal of more of the medicament in the apical third of those straight canals.     

Eradication of enterococci biofilms by lactic acid alone and combined with chlorhexidine and cetrimide

Objective: The antimicrobial activity of lactic acid (LA) alone or in combination with chlorhexidine (CHX) and cetrimide (CTR) against three Enterococcus faecalis strains, E. faecalis ATCC 29212, E. faecalis EF-D1 and E. faecalis U-1765, one Enterococcus durans strain and one dual-species biofilm was investigated. Study Design: The irrigating solutions tested were 20%, 15%, 10%, 5% and 2.5% LA, alone and in combination with 2% CHX and with 0.2% CTR. The biofilms were grown in the MBECTM high-throughput device for 24 hours and exposed to the solutions for 30 seconds and 1 minute. "Eradication" was defined as 100% bacterial kill. Results: Twenty percent LA eradicated all enterococci biofilms after 30 seconds contact time. The association of LA + 0.2% CTR achieved better results than LA alone, in contrast with the results obtained using LA + 2% CHX. E. durans was eradicated by all the tested solutions at 1 minute. The dual-species biofilm, E. faecalis ATCC 29212 + E. durans, gave intermediate values of the pure cultures. Conclusions: LA is capable of eradicating enterococci biofilm at a concentration of 20%. The combination of lower concentrations with 0.2% CTR achieved eradication after 1 minute.     

Antimicrobial substantivity of chlorhexidine-treated bovine root dentin

Previous studies have demonstrated antimicrobial substantivity in root canal dentin up to 7 days after treatment with chlorhexidine. This in vitro study assessed the antimicrobial substantivity of chlorhexidine-treated bovine root dentin over a period of 21 days. Sixty standardized bovine root sections were randomly divided into three equal groups, and their canals immersed in one of the following solutions: (i) sterile saline; (ii) 2.5% NaOCl; or (iii) 0.2% chlorhexidine (CHX). Half the specimens in each group were treated with the solution for 5 min and the other half for 7 days. After solutions were removed, the specimens were incubated at 37 degrees C in Brain Heart Infusion broth containing Enterococcus faecalis (ATCC 29212). A fresh inoculum was added to the broth every other day over a 21-day period. The canals were then enlarged with sterile burs, and the dentin shavings collected and cultured for the presence of cultivable bacteria in the dentinal tubules. Specimens treated with CHX for 7 days demonstrated significantly less dentin colonization by E. faecalis than the other specimens. CHX has potential as an intracanal medicament, if it can be applied for a period of at least 7 days.     

In vitro antimicrobial activity of phytotherapic Uncaria tomentosa against endodontic pathogens

The aim of this study was to evaluate the antimicrobial activity of Uncaria tomentosa (Willd.) DC (cat's claw) against Enterococcus faecalis, Staphylococcus aureus, and Candida albicans. Suspensions with 10(8) cells/ml of each microorganism were plated in triplicate on Mueller-Hinton agar. Wells in the agar were made and filled with 2% chlorhexidine (CHX) gel, 2% cat's claw (CC) gel, 2% CHX+CC, and 1% hydroxyethylcellulose (NAT) gel. Inhibition halos were measured after 24 h at 37°C and differences were analyzed using one-way ANOVA. The mean diameter of the microbial growth inhibition zones of 2% CHX+CC against the tested microbial strains ranged from 21.7 to 33.5 mm. This was the most effective substance against E. faecalis and C. albicans, followed by CHX and CC. Against S. aureus, CHX+CC, CHX, and CC showed similar antimicrobial activity (P > 0.05). The results indicate that all the investigated compounds had antimicrobial activity against microorganisms frequently found in infected root-filled teeth.     

Antioxidant and pro-oxidant properties of chlorhexidine and its interaction with calcium hydroxide solutions

AIM: To evaluate the antioxidant and pro-oxidant properties of chlorhexidine (CHX). METHODOLOGY: The scavenging and generation of reactive oxygen species (ROS) by CHX in the presence or absence of saturated Ca(OH)(2) solutions was evaluated. The reaction emitted chemiluminescence in the presence of lucigenin thus was determined by a luminometer to evaluate the levels of ROS production. Changes in DNA conformation were analysed by agarose gel electrophoresis. Paired Student's t-test was used to compare the difference between groups. RESULTS: Chlorhexidine (0.00002-0.02%) effectively scavenged 56-88% of the superoxide radicals generated by the xanthine/xanthine oxidase reaction. Through analysis of PUC18 DNA conformation changes, CHX was shown to be a mild scavenger of hydroxyl radicals generated by H(2)O(2) plus FeCl(2). However, CHX (>0.083%) decreased the mobility of PUC18 plasmid DNA with potential production of DNA-DNA cross-link and severe DNA breaks (presence of DNA smear) at further higher concentrations. Furthermore, CHX induced ROS production including H(2)O(2) and superoxide radicals in 0.1N NaOH (pH = 12.76) or Ca(OH)(2) (pH = 12.5) solutions. CONCLUSION: Chlorhexidine exhibited both antioxidant and pro-oxidant properties under different conditions. These events are possibly involved in the killing of root canal and periodontal microorganisms when CHX and Ca(OH)(2) were used in combination or separately. Potential genotoxicity and tissue damage when extruded into the periradicular tissue and at higher concentrations should be considered during periodontal and endodontic practice.     

Eradication of enterococci biofilms by lactic acid alone and combined with chlorhexidine and cetrimide

Objective: The antimicrobial activity of lactic acid (LA) alone or in combination with chlorhexidine (CHX) and cetrimide (CTR) against three Enterococcus faecalis strains, E. faecalis ATCC 29212, E. faecalis EF-D1 and E. faecalis U-1765, one Enterococcus durans strain and one dual-species biofilm was investigated. Study Design: The irrigating solutions tested were 20%, 15%, 10%, 5% and 2.5% LA, alone and in combination with 2% CHX and with 0.2% CTR. The biofilms were grown in the MBECTM high-throughput device for 24 hours and exposed to the solutions for 30 seconds and 1 minute. “Eradication” was defined as 100% bacterial kill. Results: Twenty percent LA eradicated all enterococci biofilms after 30 seconds contact time. The association of LA + 0.2% CTR achieved better results than LA alone, in contrast with the results obtained using LA + 2% CHX. E. durans was eradicated by all the tested solutions at 1 minute. The dual-species biofilm, E. faecalis ATCC 29212 + E. durans, gave intermediate values of the pure cultures. Conclusions: LA is capable of eradicating enterococci biofilm at a concentration of 20%. The combination of lower concentrations with 0.2% CTR achieved eradication after 1 minute. Key words:Biofilm, cetrimide, chlorhexidine, enterococcus durans, enterococcus faecalis.     

Reduction in the cultivable bacterial populations in infected root canals by a chlorhexidine-based antimicrobial protocol

The present clinical study was conducted to assess the bacterial reduction after chemomechanical preparation using 0.12% chlorhexidine digluconate solution as an irrigant and the additive antibacterial effect of intracanal dressing with calcium hydroxide (Ca(OH)(2)) associated with 0.12% chlorhexidine digluconate gel. According to stringent inclusion/exclusion criteria, 13 teeth with primary intraradicular infections and chronic apical periodontitis were selected and followed in the study. Bacterial samples were taken at the baseline (before treatment) (S1), after chemomechanical preparation using chlorhexidine (CHX) as an irrigant (S2), and after a 7-day dressing with Ca(OH)(2)/CHX paste (S3). Cultivable bacteria recovered from infected root canals at the three stages were counted and identified by means of 16S ribosomal RNA gene sequencing analysis. At S1, all canals were positive for bacteria, with the mean number of 3.5 taxa per canal (range, 2-9). At S2, 7 cases (53.8%) still harbored cultivable bacteria, with a mean number of 1.7 taxon per canal (range, 1-4). At S3, only one case (7.7%) was positive for the presence of bacteria. The great majority of taxa found in posttreatment samples were gram-positive bacteria. A significantly high reduction in bacterial counts was observed between S1 and S2 and S1 and S3 (p<0.001). Also, significant differences were observed for comparisons involving S2 and S3 samples with regard to both quantitative bacterial reduction (p=0.014) and number of cases yielding negative cultures (p=0.01). It was concluded that chemomechanical preparation with 0.12% CHX solution as an irrigant significantly reduced the number of intracanal bacteria but failed to render the canal free of cultivable bacteria in about one half of the cases. Application of a 7-day intracanal dressing with Ca(OH)(2)/CHX paste further increased significantly the number of cases yielding negative cultures.