Determination of mutagenicity of the precipitate formed by sodium hypochlorite and chlorhexidine using the Ames test

The aim of this study was to determine the direct mutagenic potential of any precipitate formed by combining sodium hypochlorite (NaOCl) and chlorhexidine (CHX). The precipitates formed by NaOCl and CHX were dissolved in 100% dimethyl sulfoxide and cultured with mutant Salmonella Typhimurium strains. The cells were observed for reverse mutation. The numbers of positive/mutated wells were statistically compared with those in the background plates using the two‐sample proportion independent t‐test. The precipitates were not found to be significantly more mutagenic than the background plates. Within the limitations of this study, the results suggest that the precipitates formed when sodium hypochlorite and chlorhexidine contact did not show mutagenic (and are therefore carcinogenic) potential.     

In vitro antimicrobial activity of several concentrations of sodium hypochlorite and chlorhexidine gluconate in the elimination of Enterococcus faecalis

AIM: The aim of this study was to assess, in vitro, the effectiveness of several concentrations of NaOCl (0.5%, 1%, 2.5%, 4% and 5.25%) and two forms of chlorhexidine gluconate (gel and liquid) in three concentrations (0.2%, 1% and 2%) in the elimination of E. faecalis. METHODOLOGY: A broth dilution test using 24-well cell culture plates was performed and the time taken for the irrigants to kill bacterial cells was recorded. Isolated 24 h colonies of pure cultures of E. faecalis grown on 10% sheep blood plus Brain Heart Infusion (BHI) agar plates were suspended in sterile 0.85% NaCI solution. The cell suspension was adjusted spectrophotometrically to match the turbidity of a McFarland 0.5 scale. One mL of each tested substance was placed on the bottom of wells of 24-well cell culture plates (Corning, NY), including the control group (sterile saline). Six wells were used for each time period and irrigant concentration. Two mL of the bacterial suspension were ultrasonically mixed for 10 s with the irrigants and placed in contact with them for 10, 30, and 45 s; 1, 3, 5, 10, 20, and 30 min; and 1 and 2 h. After each period of time, 1 mL from each well was transferred to tubes containing 2 mL of freshly prepared BHI + neutralizers in order to prevent a residual action of the irrigants. All tubes were incubated at 37 degrees C for 7 days. The tubes considered to have positive growth were those which presented medium turbidity during the incubation period. Data were analysed statistically by the Kruskal-Wallis test. with the level of significance set at P < 0.05. RESULTS: All irrigants were effective in killing E. faecalis. but at different times. Chlorhexidine in the liquid form at all concentrations tested (0.2%, 1% and 2%) and NaOCI (5.25%) were the most effective irrigants. However, the time required by 0.2% chlorhexidine liquid and 2% chlorhexidine gel to promote negative cultures was only 30 s and 1 min, respectively. CONCLUSIONS: Even though all tested irrigants possessed antibacterial activity, the time required to eliminate E. faecalis depended on the concentration and type of irrigant used.     

Evaluation of irrigant extrusion following the use of different root canal irrigation techniques: A systematic review and meta-analysis

This systematic review investigates whether different irrigation techniques have different effects on irrigant extrusion from mature tooth apices. Articles published between January 2000 and January 2022 were searched in six electronic databases (MEDLINE, Embase, Google Scholar, Web of Science, Scopus and Cochrane) using appropriate keywords. Overall, 2265 articles were screened by their titles and abstracts. Fifty-six full-text articles were selected based on the inclusion criteria. Of them, 17 in vitro studies were included in the systematic review and meta-analysis. The meta-analysis was conducted using the random-effects inverse variance method. The results showed that the negative pressure technique caused a lesser amount (p = 0.00) and frequency (p = 0.00) of extrusion than the open-ended needle irrigation. Sonic and ultrasonic activation caused less amount of extrusion than both open-ended (p = 0.00 or p = 0.01) and closed-ended needle (p = 0.00) irrigation.     

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.     

Toxicity of concentrated sodium hypochlorite used as an endodontic irrigant

AIM: To present a clinical case that illustrates the toxicity of concentrated sodium hypochlorite (NaOCl) on vital tissues. SUMMARY: The severe clinical consequences of extruding concentrated NaOCl into the periradicular tissues during root canal irrigation are described. After diagnosis of external resorption in tooth 34, endodontic treatment was initiated. Following irrigation with a concentrated NaOCl solution, a rapidly developing swelling and haematoma were visible. During the next few days, an extensive bruise and local necrosis of the oral mucosa developed. After healing of the involved tissues, the canal was prepared and obturated. At this time, no clinical symptoms remained. KEY LEARNING POINTS: * The use of concentrated NaOCl as a root canal irrigant might cause severe clinical problems when extruded into vital tissues. * The present report confirms the known toxicity of NaOCl to soft tissues following inadvertent extrusion. * To avoid extrusion, it is always prudent to confirm the length and integrity of the root canal system before irrigating with concentrated solutions.     

Neurological complications following extrusion of sodium hypochlorite solution during root canal treatment

AIM: To report the presentation and management of two cases with neurological complications secondary to the extrusion of sodium hypochlorite solution into the facial soft tissues during root canal treatment. SUMMARY: The clinical features, with particular emphasis on nerve deficit following inadvertent extrusion of sodium hypochlorite, are discussed and its management highlighted. Early and aggressive treatment is advocated following such incidents in order to reduce potentially serious complications. KEY LEARNING POINTS: *Neurological sequelae can follow inadvertent hypochlorite extrusion. *Early recognition may avert a potentially more serious outcome. *Active hospital treatment including intravenous steroids and antibiotics is recommended.     

根管冲洗剂次氯酸钠和氯亚明的细胞毒性

目的:评价次氯酸钠(NaOCl)和氯亚明溶液在相同有效氯浓度下的细胞毒性.方法: 按有效氯浓度为0.500 0%、0.250 0%、0.200 0%、0.150 0%、0.125 0%、0.062 5%分别配制NaOCl溶液和氯亚明溶液,使其与L929细胞接触2 h、30 min、10 min,用MTT法检测各组的细胞存活率.结果: 在所设定的浓度范围内,除高低两端组(0.500 0%-2 h及0.062 5%-30 min)外,细胞在其余各组NaOCl溶液中的存活率均大于氯亚明,差异有统计学意义(P<0.05).结论: 在0.250 0%以下的有效氯浓度范围,NaOCl溶液的细胞毒性小于氯亚明溶液.     

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.     

The influence of organic load on the antimicrobial activity of different concentrations of NaOCl and chlorhexidine in vitro

AIM: To evaluate bacterial growth after contact with sodium hypochlorite (NaOCl; 1 and 5%) and chlorhexidine (CHX; 0.12, 0.5 and 1%) in vitro with or without the addition of organic material (bovine serum albumin (BSA) 0.5%). METHODOLOGY: Bacterial samples (American Type Culture Collection (ATCC)) of Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Porphyromonas gingivalis and Fusobacterium nucleatum were kept in contact with each irrigating solution for varying intervals of time: immediately (t0), 5 min (t5), 15 min (t15) and 30 min (t30). Each test was repeated 10 times. In half of the specimens, 0.5% BSA was added as organic material in an attempt to simulate the organic tissue present in the root canal system. Bacterial growth under appropriate condition of incubation was evaluated and compared for each microorganism at all time intervals. RESULTS: A 0.12% CHX solution did not eliminate E. faecalis at any time interval. One percent CHX eliminated all strains, as did both NaOCl concentrations. BSA did not interfere substantially with the antimicrobial activity of any of the irrigating solutions. CONCLUSIONS: Under the condition of this study, a 0.12% CHX solution was ineffective at killing E. faecalis.     

Antibacterial activity of various root canal sealers and root-end filling materials in dentin blocks infected ex vivo with Enterococcus faecalis

Objective. The aim was to investigate the antibacterial activity of the root-end filling materials MTA and IRM, different endodontic sealers and calcium hydroxide [Ca(OH)₂] in experimentally infected dentinal tubules. Materials and methods. Ninety-four human root segments were prepared and the root canals were enlarged to ISO size 90. After smear removal, the specimens were infected with Enterococcus faecalis for 3 weeks. The roots were divided into eight groups and filled either with MTA, IRM, Ca(OH)₂, gutta-percha and EndoRez (ER)/GuttaFlow (GF)/AH Plus (AH+) or with Resilon and Epiphany (EpRe). One group of specimens was left unfilled for control. Half of the specimens were treated for 1 day and the other half for 7 days in humid conditions at 37°C. Dentin samples from each canal were collected by enlarging the canals to ISO size 150; thus a dentinal depth of 300 µm was sampled. The number of cultivable bacteria was determined for each specimen. Statistical significance was set to 5%. Results. After 1-day or 7-days of treatment, compared to control, all materials (except ER and GF at day 7) significantly reduced the number of bacteria. At day 1 and day 7, no significant difference was found between ER and GF and between Ca(OH)₂, AH+, EpRe, IRM and MTA. However, a significant difference was found between these two groups of materials (except between GF and EpRe at day 7). Significantly more bacteria were cultured in the ER, GF, EpRe and IRM groups at day 7 compared to day 1. Conclusions. All materials exerted varying degrees of antibacterial activity which generally tended to decrease with time. The most stable antibacterial effect throughout the 7-day period was for Ca(OH)₂, AH+ and MTA.     

In vitro antimicrobial and anti-endotoxin action of Zingiber Officinale as auxiliary chemical and medicament combined to calcium hydroxide and chlorhexidine

Abstract

Objective. This study was conducted in vitro to compare the effectiveness of Zingiber Officinale as an auxiliary chemical substance followed by placement of different intra-canal medication in removing endotoxins and cultivable micro-organisms from infected root canals. Materials and methods. Seventy-two root canals were contaminated with Enterococcus faecalis, Candida albicans and Escherichia coli for 28 days. After, the teeth were instrumented using Zingiber Officinale and divided into six groups according to the intra-canal medication: chlorhexidine gel; calcium hydroxide + chlorhexidine gel; glycolic ginger extract; calcium hydroxide + glycolic ginger extract; calcium hydroxide + saline solution and saline solution (control). Sample collections were performed after root canal contamination (Baseline; S1), after instrumentation (S2), 7 days after instrumentation (S3), after 14 days with intra-canal medication (S4) and 7 days after removal of intra-canal medication (S5). The results were analyzed by the Kruskal-Wallis and Dunn tests. Results. It was observed that in S2 and S3 there was significant reduction of the micro-organisms and the quantity of endotoxins after instrumentation. In samples S4 and S5 there was complete elimination of micro-organisms and significant reduction of endotoxins. Conclusion. It was concluded that Zingiber Officinale as an auxiliary chemical substance was effective on the micro-organisms tested, yet was unable to eliminate the endotoxins. Similarly, the intra-canal medication were effective on micro-organisms, yet did not completely eliminate the endotoxins.     

Antimicrobial effect of 2% sodium hypochlorite and 2% chlorhexidine tested by different methods

The objective of this study was to analyze the antimicrobial effect of 2% sodium hypochlorite (NaOCl) and 2% chlorhexidine (CHX) by agar diffusion test and by direct exposure test. Five microorganisms: Staphylococcus aureus, Enterococcus faecalis, Pseudomonas aernginosa, Bacillus subtilis, Candida albicans, and one mixture of these were used. These strains were inoculated in brain heart infusion (BHI) and incubated at 37 degrees C for 24 h. For the agar diffusion test (ADT), 18 Petri plates with 20 ml of BHI agar were inoculated with 0.1 ml of the microbial suspensions, using sterile swabs that were spread on the medium, obtaining growth injunction. Fifty-four paper disks (9 mm in diameter) were immersed in the experimental solutions for 1 min. Subsequently, three papers disks containing one of the substances were placed on the BHI agar surface in each agar plate. The plates were maintained for 1 h at room temperature, and then incubated at 37 degrees C for 48 h. The diameter of microbial inhibition was measured around the papers disks containing the substances. For the direct exposure test, 162#50 sterile absorbent paper points were immersed in the experimental suspensions for 5 min, and were then placed on Petri plates and covered with one of the irrigant solutions, or with sterile distilled water (control group). After intervals of 5, 1 0 and 30 min, the paper points were removed from contact with the solutions and individually immersed in 7 ml of Letheen Broth, followed by incubation at 37 degrees C for 48 h. Microbial growth was evaluated by turbidity of the culture medium. A 0.1 ml inoculum obtained from the Letheen Broth was transferred to 7 ml of BHI, and incubated at 37 degrees C for 48 h. Bacterial growth was again evaluated by turbidity of the culture medium. Gram stain of BHI cultures was used for verification of contamination and growth was determined by macroscopic and microscopic examination. The best performance of antimicrobial effectiveness of NaOCI was observed in the direct exposure test, and of CHX was observed in the agar diffusion test. The magnitude of antimicrobial effect was influenced by the experimental methods, biological indicators and exposure time.     

The properties and applications of chlorhexidine in endodontics

Microorganisms and their by‐products are considered to be the major cause of pulp and periradicular pathosis. Hence, a major objective in root canal treatment is to disinfect the entire root canal system, which requires that all contents of the root canal system be eliminated as possible sources of infection. This goal may be accomplished using mechanical instrumentation and chemical irrigation, in conjunction with medication of the root canal system between treatment sessions. To reduce or eliminate bacteria, various irrigation solutions have been advocated. Chlorhexidine is a cationic molecule, which can be used during treatment. It has a wide range of antimicrobial activity. Its cationic structure provides a unique property named substantivity. The purpose of this paper is to review the structure and mechanism of action of CHX, its antibacterial and antifungal activity, its effect on biofilm, its substantivity (residual antibacterial activity), its tissue solvent ability, its interaction with calcium hydroxide and sodium hypochlorite, its anticollagenolytic activity, its effect on coronal and apical leakage of bacteria, its toxicity and allergenicity and the modulating effect of dentine and root canal components on its antimicrobial activity. A Medline search was performed from 1981 to the end of March 2008 and was limited to English‐language papers. The keywords searched on Medline were ‘chlorhexidine AND endodontics’, ‘chlorhexidine AND root canal therapy’, ‘chlorhexidine AND substantivity’ and ‘chlorhexidine AND toxicity’. The reference lists of each article were manually checked for additional articles of relevance.     

Analysis of the Antimicrobial Susceptibility of Anaerobic Bacteria Isolated from Endodontic Infections in Brazil during a Period of Nine Years

Introduction

The purpose of this study was to analyze the susceptibility of some anaerobic species isolated from a Brazilian population at different periods of time by determining a pattern of development of resistance to frequently prescribed antibiotics in endodontics.

Methods

Root canal samples were collected from infected teeth at different periods of time (2000-2002, 2003-2005, and 2007-2008) and microbiologically identified with conventional culture techniques. The susceptibility of Prevotella intermedia/nigrescens, P. oralis, Fusobacterium nucleatum, and P. micra isolated strains was determined by the minimum inhibitory concentration (MIC) of amoxicillin, amoxicillin + clavulanate, benzylpenicillin, clindamycin, erythromycin, and metronidazole by using the E-test method.

Results

Amoxicillin and amoxicillin + clavulanate were effective against the majority of species at the different periods of study. Overall, there were low statistical differences regarding the microbial susceptibility between the experimental periods. However, an increase in the anaerobic resistance to penicillin G and clindamycin was observed. Resistance to erythromycin was observed in all species, and there were statistically significant differences between 2000-2002 and 2003-2005 periods for F. nucleatum (P < .05) and between 2003-2005 and 2007-2008 periods for P. intermedia/nigrescens and P. oralis (P < .05).

Conclusions

The antimicrobial resistance of anaerobes isolated from primary endodontic infections showed an increase throughout a period of time regarding a specific Brazilian population.