Establishment and maintenance of asepsis in endodontics - a review of the literature.

OBJECTIVES: Successful endodontic treatment depends on effective measures to eliminate and prevent infection of root canals. Initially treatment should start with isolation and disinfection of the operating field. This review makes an inventory of the available knowledge regarding its establishment and maintenance. MATERIALS AND METHODS: A literature search was conducted in the PubMed database in order to identify clinical trials examining disinfection or unintentional contamination of the endodontic operative field. A list of 115 articles was obtained and screened. Five relevant articles were identified. These articles were read in full text. The reference lists from these articles were checked manually for additional studies and three studies were obtained. A total of eight articles met the inclusion criteria. RESULTS: There was a great variety in terms of aim, method, and material of the included studies. None could prove a totally reliable aseptic operative field and not one chemical, or combination of chemicals, were found in more than one study. CONCLUSIONS: No study documented complete asepsis following initial disinfection, and no study could document predictable maintenance of an established bacteria-free surface. Critical appraisal and standardization of the disinfection and aseptic procedures in endodontics are needed.

Treatment of pulps in teeth affected by deep caries - A systematic review of the literature.

BACKGROUND: This systematic review assesses the effect of methods commonly used to manage the pulp in cases of deep caries lesions, and the extent the pulp chamber remains uninfected and does not cause pulpal or periapical inflammatory lesions and associated tooth-ache over time. STUDY DESIGN: An electronic literature search included the databases PubMed, EMBASE, The Cochrane Central Register of Controlled Trials and Cochrane Reviews from January 1950 to March 2013. In addition, hand searches were carried out. Two reviewers independently evaluated abstracts and full-text articles. An article was read in full if at least one of the two reviewers considered the abstract potentially relevant. Altogether, 161 articles were read in full text. Of these, 24 studies fulfilled established inclusion criteria. Based on studies of at least moderate quality, the quality of evidence of each procedure was rated in four levels according to GRADE. RESULTS: No study reached the high quality level. Twelve were of moderate quality. The overall evidence was insufficient to assess which of indirect pulp capping, stepwise excavation, direct excavation and pulp capping/partial pulpotomy, pulpotomy or pulpectomy is the most effective treatment approach for teeth with deep caries. CONCLUSIONS: Because of the lack of good studies it is not possible to determine whether an injured pulp by deep caries can be maintained or whether it should be removed and replaced with a root canal filling. Both randomized studies and prospective observational studies are needed to investigate whether a pulp exposed to deep caries is best treated by measures intended to preserve it or by pulpectomy and root filling.

Pulp exposures in adults--choice of treatment among Swedish dentists.

This study comprises a survey of Swedish dentists'treatment preferences in cases of carious exposure of the dental pulp in adults.The survey was conducted as part of a comprehensive report on methods of diagnosis and treatment in endodontics, published in 2010 by the Swedish Council on Health Technology Assessment. A questionnaire was mailed to a random subsample of 2012 dental offices where one dentist at each office was requested to answer all questions. Each questionnaire contained one of three sets of questions about endodontic practice routines.Thus around one-third of the subsample received case-specific questions about treating carious exposure. Only general practitioners aged below 70 years were included.The final study sample comprised 412 participants.The dentists were presented with two case scenarios. In Case 1 a 22-year old patient had a deep carious lesion in tooth 36 and in Case 2 a 50-year old patient had a deep carious lesion in tooth 14.The participants were asked to nominate their treatment of choice: pulp capping, partial pulpotomy or pulpectomy. For Case 1, 17 per cent of the respondents selected pulpectomy; the corresponding rate for Case 2 was 47 per cent. Female gender and age group 25-49 years were predictive of selection of less invasive treatment options. However, according to recent guidelines (2011) from the National Board of Health and Wellfare, Swedish dentists are recommended to elect pulpectomy prior to pulp capping/partial pulpotomy when confronted with a tooth having a cariously exposed pulp in adults.

Accreditation of postgraduate speciality training programmes in Endodontology. Minimum criteria for training Specialists in Endodontology within Europe.

This consensus statement from the European Society of Endodontology (ESE) sets out the minimum criteria for training Specialists in Endodontology within Europe. The case is made for recognizing Endodontology as a distinctive dental discipline throughout Europe. Guidelines are presented on the requirements of a specialist and of a specialist training programme in Endodontology. The aims, objectives and curriculum content of a specialist training pathway are outlined, with guidelines on trainee appraisal, and the expectations of faculty and institutional commitment. In publishing these guidelines, the ESE is responding to a public and professional need for consistently high standards of training and specialist clinical service within Europe.

Antibiotics Used in Regenerative Endodontics Modify Immune Response of Macrophages to Bacterial Infection.

INTRODUCTION: Ciprofloxacin, amoxicillin, and metronidazole are antibiotics used in regenerative endodontic therapy (RET). Although their antimicrobial properties are well-documented, there is a lack of information on the effects of these antibiotics on the immune response by host macrophages and periapical healing. Thus, this study had 2 objectives: (1) to determine the immune response of macrophages to bacterial infection in response to the combination of ciprofloxacin or amoxicillin and metronidazole and (2) using conditioned media produced by these macrophages to simulate the periapical microenvironment, to determine the impact on the expression of extracellular matrix (ECM) components by periodontal fibroblasts. METHODS: Macrophages were treated with ciprofloxacin and metronidazole or amoxicillin and metronidazole at 10-1000 µg/mL. The treated macrophages were exposed to lipopolysaccharide, and the pro- and anti-inflammatory cytokines produced were quantified with enzyme-linked immunosorbent assay. Periodontal fibroblasts were treated with conditioned media from these treated macrophages, and the expression of ECM genes was determined by quantitative polymerase chain reaction. RESULTS: Lipopolysaccharides elicited the production of proinflammatory cytokines interleukin 1 beta and tumor necrosis factor alpha by macrophages, but this was suppressed by ciprofloxacin and metronidazole. Moreover, only conditioned media from macrophages treated with ciprofloxacin and metronidazole rescued microbial-induced down-regulation of ECM genes by periodontal fibroblasts. Specifically, ciprofloxacin was the antibiotic responsible for these observations. In contrast, these effects were not observed with amoxicillin and metronidazole. CONCLUSIONS: Apart from disinfection of the root canal system, the combination of ciprofloxacin and metronidazole also exerts an immunomodulatory effect, which may aid in periapical healing.

Rapid method for the detection of root canal bacteria in endodontic therapy.

INTRODUCTION: Complete eradication of microorganisms is essential for successful root canal therapy. However, current methods to evaluate persistent bacteria after therapy are not widely practiced. Adenosine triphosphate (ATP) is an indicator of viable cells. The bioluminescence-based ATP assay is easy to perform, and results can be obtained in a clinically relevant time frame of 5 minutes. The aims of this study were to evaluate the sensitivity of the ATP detection method and the specificity of this assay for viable cells and to compare the ATP and culture methods from root canal samples of patients undergoing endodontic treatment. METHODS: The sensitivity of the ATP assay was determined using bacterial species commonly isolated from root canals. Bacteria were treated with sodium hypochlorite; after which, culture plating and the ATP assay were performed. Forty-three root canal samples before (S1) and after (S2) instrumentation and 36 samples after the removal of calcium hydroxide dressing (S3) were collected from patients undergoing root canal treatment and subjected to ATP assay and anaerobic culture. RESULTS: The sensitivity of the ATP assay was determined to be between 10 and 100 bacterial cells. This method of detection also correlated well with the presence of viable bacteria. The ATP readings obtained allowed clear segregation of anaerobic culture-positive and -negative samples obtained from infected root canals of patients. CONCLUSIONS: The ATP detection method can be used as a rapid tool to determine the presence of viable bacteria during root canal therapy. This method may be potentially useful as an adjunct to root canal treatment.

Isolation of alkaline-tolerant bacteria from primary infected root canals.

INTRODUCTION: Alkaline-tolerant bacteria in primary infected root canals could have enhanced survival capacity against antimicrobials commonly used in root canal treatment. The aims of this study were to isolate and characterize alkaline-tolerant bacteria before endodontic treatment (S1), after chemomechanical root canal preparation (S2), and after calcium hydroxide dressing (S3). METHODS: Bacteriologic samples were obtained from 43 primary infected root canals. Samples were inoculated into culture media at a pH of 9 and incubated anaerobically. The identities of bacterial isolates were determined by 16S ribosomal RNA sequencing. RESULTS: All S1 samples were culture positive, with 70% harboring bacteria tolerating a pH of 9. Gram-positive bacteria Pseudoramibacter alactolyticus and Streptococcus spp were the most frequently isolated strains with a prevalence of 54%. Of 13 culture-positive S2 samples, 8 isolates tolerated a pH of 9, namely Streptococcus sanguinis, Enterococcus faecalis, Enterobacter cancerogenus, Streptococcus oralis, and Fusobacterium nucleatum. Seven of these 8 isolates (88%) were correspondingly isolated at S1. All 3 culture-positive S3 samples tolerated a pH of 9, namely S. sanguinis and E. faecalis, which were also isolated in the corresponding S1 and S2 samples. CONCLUSIONS: We showed that the presence of alkaline-tolerant Streptococcus and Enterococcus spp in primary infected root canals could lead to their persistence during and after root canal treatment and could pose a challenge to current treatment efficacy.

The effects of antimicrobials on endodontic biofilm bacteria.

INTRODUCTION: In the present study, confocal microscopy, a miniflow cell system, and image analysis were combined to test in situ the effect of antimicrobials and alkali on biofilms of Enterococcus faecalis, Lactobacillus paracasei, Streptococcus anginosus, and Streptococcus gordonii isolated from root canals with persistent infections. METHODS: Biofilms formed for 24 hours were exposed for 5 minutes to alkali (pH = 12), chlorhexidine digluconate (2.5%), EDTA (50 mmol/L), and sodium hypochlorite (1%). The biofilms were then characterized by using fluorescent markers targeting cell membrane integrity (LIVE/DEAD) and metabolic activity (5-cyano-2,3-ditolyl tetrazolium chloride and fluorescein diacetate). In addition, the biofilm architecture and the extent to which coating of the substrate surface with collagen influenced the resistance pattern to the chemicals were also analyzed. RESULTS: NaOCl (1%) affected the membrane integrity of all organisms and removed most biofilm cells. Exposure to EDTA (50 mmol/L) affected the membrane integrity in all organisms but failed to remove more than a few cells in biofilms of E. faecalis, L. paracasei, and S. anginosus. Chlorhexidine (2.5%) had a mild effect on the membrane integrity of E. faecalis and removed only 50% of its biofilm cells The effects were substratum-dependent, and most organisms displayed increased resistance to the antimicrobials on collagen-coated surfaces. CONCLUSIONS: The biofilm system developed here was sensitive and differences in cell membrane integrity and removal of biofilm cells after exposure to antimicrobials commonly used in endodontics was discernible.

Incidence of periapical lesions and clinical symptoms after pulpectomy--a clinical and radiographic evaluation of 1- versus 2-session treatment.

OBJECTIVE: Outcome of pulpectomy in 2 treatment sessions with calcium hydroxide as an intracanal dressing was compared to a procedure comprising instrumentation and root-filling in 1 session. STUDY DESIGN: Subjects with a vital pulp condition (N = 256) were recruited to a randomized clinical trial. Outcome parameters included radiographic signs of apical periodontitis and painful symptoms at clinical follow-ups 1 week and 1-3 years after treatment. RESULTS: Of 244 subjects available for final recall, 17 presented with periapical radiolucency. Lesions were evenly distributed among the 2 treatment groups. Postoperative pain recorded 1 week after permanent filling was significantly associated with overfilling (P = .001), with no difference between treatment groups. There was no association with presence of overfilling and radiographic lesion at end point of recall. CONCLUSIONS: Study confirms that pulpectomy may be carried out at a high rate of success if due attention is given to aseptic operating procedures, proper instrumentation and filling. Under these conditions an interappointment dressing with calcium hydroxide does not seem to influence outcome.

Observations on healing following endodontic surgery in nonhuman primates (Macaca fascicularis): effects of rhBMP-2.

OBJECTIVES: The potential of recombinant human bone morphogenetic protein-2 (rhBMP-2) to enhance bone healing following endodontic surgery was tested. The pattern and timing of de novo bone formation and cementum regeneration, and the potential for root resorption and ankylosis to accompany bone formation were evaluated. STUDY DESIGN: Pulpal infections were induced in maxillary and mandibular incisor teeth in young adult Cynomolgus monkeys. The teeth received conventional endodontic treatment immediately followed by surgical root resection. In a randomized split-mouth design, contralateral apical bone defects received rhBMP-2 in absorbable collagen sponge (ACS) carrier or served as sham-surgery controls to provide histological and radiographic evaluations following 1 (mandibular incisors) and 4.5 (maxillary incisors) month(s) postsurgery. RESULTS: At 1 month postsurgery trabecular bone filled the apical bone defects. The newly formed bone appeared considerably more mature and had assumed characteristics of the contiguous resident bone at 4.5 months postsurgery. The resected root tips were almost completely covered by new cementum with a maturing functionally oriented periodontal ligament. Localized inflammatory infiltrates were associated with the filled root canals and extruded root-filling material. Root resorption and ankylosis were not observed. There were no apparent differences in healing patterns between sites implanted with rhBMP-2/ACS and those serving as sham-surgery controls. CONCLUSIONS: Under conditions where the influence of infectious elements and irritation caused by root filling material are minimized, bone formation and cementum regeneration appears rapid following endodontic surgery. rhBMP-2/ACS did not offer an obvious benefit above and beyond that of the native osteogenic potential in this animal model.

Advances since the paper by Zander and Glass (1949) on the pursuit of healing methods for pulpal exposures: historical perspectives.

Clinical observations and experimental studies in humans and laboratory animals have demonstrated that healing and repair of pulpal exposures by caries, trauma, or iatrogenic causes are possible with a variety of wound treatment methods. Yet clinical trials have shown that predictable long-term pulp tissue preservation may be an elusive goal and has led to doubts about pulp capping and pulpotomy as valid clinical procedures. Nevertheless substantial knowledge has accumulated over the years on the mechanisms and the treatment factors that are important to promote/support continued vital pulp functions. This article highlights some key contributions to our current knowledge base, which have come to light during the more than 50 years since a pioneering experimental study by Zander and Glass was published in the Triple O journal.

Streptococci from root canals in teeth with apical periodontitis receiving endodontic treatment.

OBJECTIVES: The object of this study was to investigate the diversity among streptococcal species isolated from root canals in conjunction with endodontic therapy and to characterize their production of extracellular proteins. STUDY DESIGN: Consecutive root canal samples (RCS) taken as bacteriological controls during root canal treatment of teeth with apical periodontitis were analyzed in a total of 100 clinical cases. Bacteria were isolated and classified by selective media and gas liquid chromatography. Streptococcal strains were identified by carbohydrate fermentation, hydrolysis of aesculin/arginine, and production of enzymes. Releases of extracellular proteins by streptococci and Enterococcus spp in fluid culture media were examined with SDS-PAGE and 2-dimension gel electrophoresis (2 DE). Extracellular proteins produced were quantified and qualitatively analyzed. Specific proteins were targeted with Western immunoblot assays. Comparisons were made with type strains. RESULTS: Of a total of 241 bacterial strains recovered in the first samples submitted, Streptococcus gordonii, S anginosus, and S oralis were the most frequently isolated streptococci. In 49 of 89 resubmitted samples showing bacterial growth, S gordonii and S oralis still predominated among streptococci. Other common bacterial isolates were Enterococcus spp, Lactobacillus paracasei, and Olsenella uli. Quantitative and qualitative differences in extracellular protein production were observed among clinical isolates and laboratory streptococcal strains. In similar conditions for growth, S intermedius, S anginosus, S oralis, and S gordonii were strong producers of extracellular proteins (>3.0 microg/mL), while Enterococcus spp and S mutans were weak. Whole cell protein extracts showed a different profile from that of extracellular proteins. The chaperone protein DnaK was recognized to be produced extracellularly by S gordonii, S oralis, S anginosus, and S parasanguis. CONCLUSIONS: Being strong producers of extracellular proteins and by virtue of common presence in teeth undergoing endodontic therapy, S gordonii, S anginosus, and S oralis may be of pathogenic significance in posttreatment apical periodontitis.