[The assessment of the endodontic foundation]. / De beoordeling van het endodontische fundament.

The diagnosis of an endodontic disease requires thorough research, collecting both clinical and radiographic information. The clinical examination includes history taking, visual inspection of the tooth and surrounding tissues, palpation of the soft and hard tissues, periodontal examination and percussion. The radiographic examination provides valuable information, but can never stand alone in arriving at a diagnosis. It is important to link the findings of the radiographic examination to other information. Sometimes, invasive examination is necessary, during which the coronal restoration is removed to allow better assessment of the tooth. This can provide additional information about the presence of caries, fractures, leakage of the restoration or other reasons for failure of the initial root canal treatment. A good diagnosis is essential for planning successful follow-up treatment.

Diffusion of antimicrobials in multispecies biofilms evaluated in a new biofilm model.

AIM: To describe the application of a newly-developed in vitro model in which the diffusion of antimicrobials in oral biofilms can be studied. METHODOLOGY: In a flow chamber consisting of three parallel feeding channels connected with each other by eight perpendicular side channels, multispecies biofilms were grown from saliva of a single donor for 48 h. The dimensions of the side channels were 100 µm × 100 µm × 5130 µm (H × W × L). When one or more side channels were filled with biofilm, the biofilms were stained with fluorescent stains. Then, one side-channel biofilm was selected and treated with phosphate buffered saline, 2% sodium hypochlorite (NaOCl), 17% ethylenediaminetetra-acetic acid (EDTA) or modified salt solution (MSS). Diffusion of the irrigants was observed by acquiring fluorescence images at 10× objective every 15 s for 30 min. RESULTS: It was possible to culture biofilms in the narrow (100 µm) channels. The biofilms varied in phenotype. In this model, no diffusion of NaOCl into the biofilms was seen after its application. Seventeen-percentage EDTA only diffused into the biofilm up to 200 µm in 30 min. MSS did diffuse in the biofilm over a distance of 450 µm within 2 min after a single application. CONCLUSIONS: This new model enables the investigation of the diffusion of antimicrobials in biofilms. Other applications to improve our understanding of the characteristics of biofilms are now possible.

In mixed biofilms Enterococcus faecalis benefits from a calcium hydroxide challenge and culturing.

AIM: To evaluate the fate of Enterococcus faecalis in dual-species and multispecies biofilms after treatment with calcium hydroxide (Ca(OH)2 ). METHODOLOGY: Biofilms were cultured from laboratory strains of E. faecalis and Pseudomonas aeruginosa, or from microbiota retrieved from primary root canal infections. The biofilms were then treated with Ca(OH)2 . The proportion of E. faecalis and P. aeruginosa and their susceptibility to disinfection were evaluated in a viability assay. In the mixed-species assay, the presence and proportions of E. faecalis before and after Ca(OH)2 treatment were evaluated with a quantitative polymerase chain reaction assay. Groups were compared using Mann-Whitney U-test and Student's t-tests. An α <0.05 was considered statistically significant. RESULTS: After Ca(OH)2 treatment in dual-species biofilms, the proportion of E. faecalis had increased (P < 0.001), whereas the total number of CFUs per biofilm was equal. Enterococcus faecalis was equally susceptible to disinfection by sodium hypochlorite (NaOCl) or by chlorhexidine. Pseudomonas aeruginosa had become more susceptible to NaOCl disinfection. The root canal isolates contained no detectable amounts of E. faecalis. After biofilm culturing or Ca(OH)2 treatment, it appeared that E. faecalis must have been present in 5 of 6 (83%) root canal samples. CONCLUSIONS: Calcium hydroxide favours the population of E. faecalis in a dual-species biofilm. Culturing multispecies root canal isolates makes E. faecalis detectable. E. faecalis was often present in primary endodontic infections, albeit in low numbers.

Cytotoxicity, interaction with dentine and efficacy on multispecies biofilms of a modified salt solution intended for endodontic disinfection in a new in vitro biofilm model.

AIM: To investigate the cytotoxicity of a modified salt solution (MSS) and evaluate the antimicrobial properties of MSS on in vitro biofilm models. METHODOLOGY: In a metabolic assay, fibroblasts derived from periodontal ligaments (PDL) of human extracted teeth were cultured and challenged with MSS or controls. Then, in active attachment biofilm models, the efficacy of MSS in the presence of dentine powder and in eliminating mature biofilms was investigated. In the dentine assay, a biofilm of Enterococcus faecalis was employed. For the final assay, microorganisms were retrieved from infected root canals and cultured to produce biofilms. After the treatments with MSS or the controls, the biofilms were collected, serially diluted and plated. The colony-forming units were counted. One-way anova was used to analyse the differences between the groups. A P < 0.05 was considered significant. RESULTS: The PDL fibroblasts remained metabolically active after challenges with MSS. Dentine powder did not alter the efficacy of MSS (P > 0.05). In endodontic biofilms, the culturable bacteria were equally reduced by MSS, 2% chlorhexidine (CHX) or 2% sodium hypochlorite (NaOCl) (P > 0.05). CONCLUSIONS: Modified salt solution is noncytotoxic in vitro and has good antimicrobial properties equal to CHX and NaOCl. Although the results are promising, ex vivo and in vivo studies are needed before its use as an interappointment root canal dressing can be considered.

[Endodontics in motion: new concepts, materials and techniques 4. Root canal disinfection in 2015]. / Endodontologie in beweging: nieuwe concepten, materialen en technieken 4. Wortelkanaaldesinfectie anno 2015.

Apical periodontitis is an inflammatory response around the root tip of a tooth to microbial infection of the root canal system. Therefore, disinfection of the root canal system is the most important aim of root canal treatment. There are various mechanical and chemical ways to clean and disinfect. Most methods, however, cannot be relied upon to fully decontaminate in all cases. There are problems, for example, with the proper concentrations of disinfectant agents, like sodium hypochlorite. But the more recent agents, like ethylenediaminetetraacetic acid, calcium hydroxide or antibiotic pastes also have disadvantages, which are mostly a result of poor access of the irrigant to the biofilm bacteria in the affected root canals. Currently, a new strategy with a modified salt solution is under investigation that offers the prospect of being used as a root canal irrigant. At this moment the preferred treatment still seems to be to remove infected tissue as much as possible and to create access for irrigation procedures. The best results are achieved with 1-2% sodium hypochlorite as a disinfectant, possibly alternating with ethylenediaminetetraacetic acid as a cleansing agent. There is no scientific evidence for the successful use of calcium hydroxide.

The Effect of Unintentional AH-Plus Sealer Extrusion on Resolution of Apical Periodontitis After Root Canal Treatment and Retreatment-A Retrospective Case-control Study.

INTRODUCTION: This study aimed to investigate the effect of unintentional AH Plus sealer extrusion on the outcome of nonsurgical root canal treatment and retreatment in teeth with apical periodontitis (AP) compared to teeth with AP and no sealer extrusion at the 1-year follow-up. METHODS: In a retrospective case-control study conducted from September 2016 to September 2021, teeth presenting clinical and radiographic signs of AP prior to treatment were included. The study comprised 2 groups: AH Plus sealer extrusion group (n = 60): initial root canal treatment or nonsurgical retreatment with unintentional apical extrusion of AH Plus sealer; and no AH Plus sealer extrusion group (n = 60): underwent the same treatment types without sealer extrusion. Additional factors, including sex, age, tooth type, tooth location, and treatment type, were recorded as potential outcome predictors. At 1-year follow-up, teeth were evaluated clinically and radiographically, and outcomes were classified as favorable (healed/healing) or unfavorable (uncertain/non-healing). Statistical analyses, such as the χ2 test, logistic regression, and Mann-Whitney U test, were employed for data analysis. RESULTS: A favorable outcome was observed in 88.4% and 85% of the cases of the AH Plus sealer extrusion and no AH Plus sealer extrusion groups, respectively. There was no statistically significant difference between the groups in terms of outcomes (P > .05) when analyzed categorically or ordinally. Of independent variables, nonsurgical retreatment showed lower healing of AP in both groups. CONCLUSIONS: Unintentional AH Plus sealer extrusion did not affect the healing of AP. However, initial root canal treatment showed better healing of AP than nonsurgical retreatment in the presence of AH Plus sealer extrusion. The analyzed outcome predictors had no effect on treatment outcome.

The effects of hyperosmosis or high pH on a dual-species biofilm of Enterococcus faecalis and Pseudomonas aeruginosa: an in vitro study.

AIM: To investigate the effect of hyperosmotic hyperosmosis or alkaline stress on a dual-species biofilm of Enterococcus faecalis and Pseudomonas aeruginosa. METHODOLOGY: Biofilms were grown on glass cover slips suspended in bacterial inoculate for 96 h, after which the cover slips with attached biofilms were immersed in brain heart infusion broth (BHI-broth) with 6 mol L(-1) sodium chloride (NaCl) representing the hyperosmotic group or Ca(OH)(2), pH 12.1, representing the alkaline group. Two per cent sodium hypochlorite and BHI- broth served as positive and negative controls, respectively. After treatment, the biofilms were washed, harvested and plated on blood-agar plates after serial dilution. The bactericidal effect was assessed by determining the colony-forming units (CFU). The effect on the biofilm mass was imaged with confocal laser scanning microscopy (CLSM). RESULTS: Hyperosmosis reduced the CFU of both species significantly after 72 h (P < 0.0001). After 168 h, P. aeruginosa was eradicated and the E. faecalis reduction was more than 99%. High pH could not induce a significant bacterial reduction. CLSM revealed dense flocculation of the biofilms incubated in alkaline broth. CONCLUSION: Hyperosmosis effectively reduced a dual-species biofilm of E. faecalis and P. aeruginosa, whilst high pH had limited bactericidal effect in this model.

The influence of apical periodontitis on the concentration of inflammatory mediators in peripheral blood plasma and the metagenomic profiling of endodontic infections: Study design and protocol.

Increased systemic inflammation has been identified in presence of oral disease, specifically endodontic disease. It is important to investigate whether treatment of the oral disease ameliorates systemic inflammation. Furthermore, there is no information about the extent to which different microorganisms may trigger inflammatory response. OBJECTIVES: Primarily (i) to compare the plasma concentrations of inflammatory mediators of apical periodontitis (AP) subjects to controls, (ii) to evaluate whether elimination of the endodontic infection reduces systemic inflammation (iii) to investigate the microbiome of root canal infections. Secondarily i) to correlate the inflammatory mediator data with the microbiome data to investigate whether the type of infection influences the type and severity of the inflammatory condition ii) to examine patterns in the inflammatory mediator data before and after tooth extraction in order to establish a biomarker signature of AP/oral disease.This is a multi-centre prospective case-control intervention study. The cohort will consist of 30 healthy human volunteers with one or two teeth with a root-tip inflammation and 30 matched healthy controls. Peripheral blood will be drawn at 6 time points, 3 before and 3 after the extraction of the tooth with apical periodontitis. The teeth will be pulverized, DNA extraction and sequencing will be performed.This study aims to compare the concentration of inflammatory blood plasma proteins in between AP-subjects and controls at different time points before and after the tooth extraction in a systematic and complete way. Additionally the composition of the root canal microbiome in association with the inflammatory response of the host will be assessed.

Reactive oxygen species can be traced locally and systemically in apical periodontitis: A systematic review.

OBJECTIVES: The aim of this systematic review was to summarize the existing evidence on the local production and systemic traces of reactive oxygen species (ROS) in apical periodontitis (AP). DESIGN: A search of MEDLINE-PubMed and EMBASE was conducted up to January 12 of 2021 to identify studies in 6 different languages. Eligibility was evaluated and data were extracted from the eligible studies following the predefined objective. The Newcastle-Ottawa Scale was used for quality assessment of the included studies. RESULTS: After screening, 21 papers met the inclusion criteria. Six studies were about systemic oxidative stress, 14 studies examined local production of reactive oxygen species and one studied both. ROS modulate cell signalling and cause oxidant imbalance locally at the site of AP. Cell signalling leads to a pro-inflammatory response, activation of MMPs and formation and progression of the AP lesion. Simultaneously, these oxidative stress biomarkers are also found in blood and saliva of subjects with AP. CONCLUSIONS: Understanding the mechanism of ROS generation, involved in chronic inflammation, can provide us with important information to enhance local and systemic healing and possibly improve diagnostic tools. Future research considerations would be to use antioxidants to accelerate the return to oxidative balance.

Does apical periodontitis have systemic consequences? The need for well-planned and carefully conducted clinical studies.

Apical periodontitis, infection of the root canal system, may have systemic consequences. This proposal has been brought forward many times in dentistry literature but the general consensus is that there is no scientific basis for an association between endodontic infections and general health. This opinion paper argues that, in order to obtain such a scientific basis, or to rule out the issue all together, we need carefully designed longitudinal challenge model (that is, intervention) studies in which we follow specific biomarkers of inflammation. These biomarkers can be those that are currently being substantiated in chronic inflammation and low-grade inflammation studies in medicine and nutritional science, where the presence of these inflammatory disorders is linked to systemic outcomes. A list of suggested biomarkers has been included.