PRIASE 2021 guidelines for reporting animal studies in Endodontology: explanation and elaboration.

Laws and ethics require that before conducting human clinical trials, a new material, device or drug may have to undergo testing in animals in order to minimize health risks to humans, unless suitable supporting grandfather data already exist. The Preferred Reporting Items for Animal Studies in Endodontology (PRIASE) 2021 guidelines were developed exclusively for the specialty of Endodontology by integrating and adapting the ARRIVE (Animals in Research: Reporting In Vivo Experiments) guidelines and the Clinical and Laboratory Images in Publications (CLIP) principles using a validated consensus-based methodology. Implementation of the PRIASE 2021 guidelines will reduce potential sources of bias and thus improve the quality, accuracy, reproducibility, completeness and transparency of reports describing animal studies in Endodontology. The PRIASE 2021 guidelines consist of a checklist with 11 domains and 43 individual items and a flowchart. The aim of the current document is to provide an explanation for each item in the PRIASE 2021 checklist and flowchart and is supplemented with examples from the literature in order for readers to understand their significance and to provide usage guidance. A link to the PRIASE 2021 explanation and elaboration document and PRIASE 2021 checklist and flowchart is available on the Preferred Reporting Items for study Designs in Endodontology (PRIDE) website (http://pride-endodonticguidelines.org/priase/).

PRIASE 2021 guidelines for reporting animal studies in Endodontology: a consensus-based development.

Animal testing is crucial in situations when research on humans is not allowed because of unknown health risks and ethical concerns. The current project aims to develop reporting guidelines exclusively for animal studies in Endodontology, using an established consensus-based methodology. The guidelines have been named: Preferred Reporting Items for Animal Studies in Endodontology (PRIASE) 2021. Nine individuals (PD, VN, AK, PM, MN, JF, EP, JJ and SJ), including the project leaders (PD, VN) formed a steering committee. The steering committee developed a novel checklist by adapting and integrating their animal testing and peer review experience with the Animals in Research: Reporting In Vivo Experiments (ARRIVE) guidelines and also the Clinical and Laboratory Images in Publications (CLIP) principles. A PRIASE Delphi Group (PDG) and PRIASE Online Meeting Group (POMG) were also formed. Thirty-one PDG members participated in the online Delphi process and achieved consensus on the checklist items and flowchart that were used to formulate the PRIASE guidelines. The novel PRIASE 2021 guidelines were discussed with the POMG on 9 September 2020 via a Zoom online video call attended by 21 individuals from across the globe and seven steering committee members. Following the discussions, the guidelines were modified and then piloted by several authors whilst writing a manuscript involving research on animals. The PRIASE 2021 guidelines are a checklist consisting of 11 domains and 43 individual items together with a flowchart. The PRIASE 2021 guidelines are focused on improving the methodological principles, reproducibility and quality of animal studies in order to enhance their reliability as well as repeatability to estimate the effects of endodontic treatments and usefulness for guiding future clinical studies on humans.

Electrokinetic transport and distribution of antibacterial nanoparticles for endodontic disinfection.

AIM: To assess a novel, noninvasive intervention capable of mobilizing charged antibacterial nanoparticles to the apical portions of the root canal system, utilizing the principles of electrokinetics. METHODS: Experiments were conducted in three stages. Stage-1: A computer model was generated to predict and visualize the electric field and current density distribution generated by the proposed intervention. Stage-2: Transport of chitosan nanoparticles (CSnp) was evaluated qualitatively using a transparent microfluidic model with fluorescent-labelled CSnp. Stage-3: An ex vivo model was utilized to study the antimicrobial efficacy of the proposed treatment against 3-week-old monospecies E. faecalis biofilms. Scanning electron microscopy (SEM) was also utilized in this stage to confirm the deposition of CSnp. RESULTS: The results of the computer simulations predicted an electric field and current density that reach their maxima at the apical constriction of the root canal. Correspondingly, the microfluidic experiments demonstrated rapid, controlled CSnp transport throughout the simulated root canal anatomy with subsequent distribution and deposition in the apical constriction as well as periapical regions. Infected root canals when subjected to the novel treatment method resulted in a mean bacterial reduction of 2.1 log CFU. SEM analysis revealed electrophoretic deposition of chitosan nanoparticles onto the root canal dentine walls in the apical region. CONCLUSION: The findings from this study demonstrate that the combination of cationic antibacterial nanoparticles with a low-intensity electric field results in particle transportation (electrophoresis) and deposition within the root canal. This results in a synergistic antibiofilm efficacy and has the potential to enhance root canal disinfection.

Efficacy of bioactive nanoparticles on tissue-endotoxin induced suppression of stem cell viability, migration and differentiation.

AIM: To characterize a lipopolysaccharide (LPS)-treated dentine tissue model (LPS dentine) to analyse the efficacy of polycationic chitosan nanoparticles (CSnp) and/or dexamethasone conjugate chitosan nanoparticles (Dex-CSnp) on the viability/differentiation potential of stem cells from apical papilla (SCAP) when exposed to LPS dentine. A further aim was to understand the effect of macrophage-dependent inflammation on SCAP migration in the presence of LPS dentine. METHODOLOGY: A total of 88 dentine slabs were used. TOF-SIMS analysis was performed amongst the LPS-treated and untreated dentine groups (n = 2/group). The study was conducted using four dentine groups: no treatment (control); LPS treatment only; LPS treatment followed by CSnp conditioning; and LPS treatment followed by Dex-CSnp conditioning groups. SCAP adherence, viability, differentiation and biomineralization potential on dentine from different groups were studied using fluorescent and scanning electron microscopy. Inflammation by macrophages in response to LPS dentine was quantified, and effect on SCAP migration was analysed. Statistical analysis was performed using Student's t-test with a significance level of P < 0.05. RESULT: TOF-SIMS analysis confirmed LPS contamination. LPS dentine affected SCAP viability but not adherence to dentine (P < 0.001). Conditioning of LPS dentine with either nanoparticles improved SCAP viability (P < 0.01) and rescued other LPS related adverse effects on SCAPs, such as F-actin disruption, decrease in differentiation/biomineralization potential. IL-6 produced by macrophages in response to LPS-treated dentine impeded SCAP migration (P < 0.001), diminished on CSnp and Dex-CSnp conditioning groups (P < 0.01). CONCLUSION: This study developed an LPS-dentine model and highlighted the ability of CSnp and Dex-CSnp to promote stem cell viability, migration, differentiation potential and reduce inflammation, providing an environment conducive for tissue regeneration/repair.

Alternative model for cathepsin K activation in human dentin.

OBJECTIVE: To evaluate the protease activity in dentin matrices subjected to lactic acid (LA) in comparison to polyacrylic acid (PAA) challenge model at cathepsin K (CT-K) optimum pH 5.5 to assess effectiveness of inhibitors in dentin collagen degradation. METHODS: Dentin disks measuring 0.5mm prepared from human molars were completely demineralized in 10% H3PO4. Demineralized dentin disks were challenged with 0.1M LA, 1.1mM PAA, artificial saliva (AS), or deionized water (C) for 24h or 7-days. Dentin collagen properties were tested by measurement of %dry mass change, and ultimate tensile strength (UTS). Degradation of dentin type I collagen was measured by telopeptide assays measuring the sub-product release of C-terminal cross-linked telopeptides (ICTP) and C-terminal peptide (CTX) in the incubation media in relation to total protein concentration, which correlates with matrix metalloproteinases (MMPs) and CT-K activities. RESULTS: Gravimetric analysis showed statistically significant difference between C and other groups (p<0.04) at 24h. LA specimens showed significantly higher weight loss from 24h to 7-days (p=0.02). UTS revealed statistically significant difference between AS and LA at 24h and 7-days. UTS at 24h and 7-days for C and AS had significantly higher mean values compared to LA and PAA. Telopeptide assays reported that CTXtp results showed that LA at 24h had significantly higher mean values compared to C and AS. SIGNIFICANCE: LA has the ability to activate endogenous CT-K in dentin as measured by the release of CTX (CT-K specific telopeptide). This LA based model has the potential application for further investigations on the activity and possible inhibitors of CT-K in human dentin.

Preferred Reporting Items for Animal Studies in Endodontology: a development protocol.

The regulated use of animals in endodontic research is often necessary to investigate the biological mechanisms of endodontic diseases and to measure the preclinical efficacy, biocompatibility, toxicology and safety of new treatments, biomaterials, sealers, drugs, disinfectants, irrigants, devices and instruments. Animal testing is most crucial in situations when research on humans is not ethical, practical or has unknown health risks. Currently, there is a wide variability in the quality of manuscripts that report the results of animal studies. Towards the goal of improving the quality of publications, guidelines for preventing disability, pain, and suffering to animals, and enhanced reporting requirements for animal research have been developed. These guidelines are referred to as Animals in Research: Reporting In Vivo Experiments (ARRIVE). Henceforth, causing any form of animal suffering for research purposes is not acceptable and cannot be justified under any circumstances. The present report describes a protocol for the development of welfare and reporting guidelines for animal studies conducted in the specialty of Endodontology: the Preferred Reporting Items for Animal Studies in Endodontology (PRIASE) guidelines. The PRIASE guidelines will be developed by adapting and modifying the ARRIVE guidelines and the Clinical and Laboratory Images in Publication (CLIP) principles. The development of the new PRIASE guidelines will include a five-step consensus process. An initial draft of the PRIASE guidelines will be developed by a steering committee. Each item in the draft guidelines will then be evaluated by members of a PRIASE Delphi Group (PDG) for its clarity using a dichotomous scale (yes or no) and suitability for its inclusion using a 9-point Likert scale. The online surveys will continue until each item achieves this standard, and a set of items are agreed for further analysis by a PRIASE Face-to-face Consensus Meeting Group (PFCMG). Following the consensus meeting, the steering committee will finalize and confirm the PRIASE guidelines taking into account the responses and comments of the PFCMG. The PRIASE guidelines will be published and disseminated internationally and updated periodically based on feedback from stakeholders.

Impact of apical extent of root canal filling on vertical root fracture: a case-control study.

AIM: To investigate the impact of apical extent of root filling on vertical root fracture (VRF) in a case-control study. METHODOLOGY: Eighty-six patients (119 roots) diagnosed with VRF in crowned root filled anterior and posterior teeth were selected. The cases were matched individually with control teeth in a ratio of 1:1 for age (±5 years), gender, tooth type, canal instrumentation method, master apical file (MAF) size and taper, technique of canal filling and time period after root filling. All root canals had been prepared using nickel-titanium (NiTi) rotary instruments and filled using the lateral compaction technique. The apical extent of root filling (overfilled to or beyond the radiographic apex or not overfilled and short of the radiographic apex) was recorded as the dependent variable by two individual examiners. Inter-examiner agreement was obtained using Kappa statistics. Recorded numbers of overfilled and not overfilled canals in cases and controls were analysed using chi-square tests and conditional logistic regression, and odds ratio was calculated. In addition, the frequency distribution of vertical and cross-sectional extensions and the course of VRFs were evaluated. RESULTS: The mean age of patients with VRFs was 50 ± 10 years with 27 (31%) males and 59 (69%) females. The Kappa score for inter-observer agreement was 0.832 (P < 0.001). There was a significant difference between cases and controls with respect to apical extent of root filling (P < 0.0001). When compared to roots not overfilled, overfilled roots had 11.5 times higher odds for occurrence of VRF (OR = 11.5; CI: 4.99 - 26.48). Most VRFs had a complete corono-apical longitudinal extension and were present bucco-lingually/palatally. CONCLUSION: After matching for age, gender, tooth type, MAF size and taper, canal filling technique and time period after root filling, root canals filled to or beyond the radiographic apex following lateral compaction had a greater association with VRF than canals filled short of the radiographic apex.

Inflammatory potential of monospecies biofilm matrix components.

AIM: To assess the inflammatory potential of biofilm matrix constituents of Enterococcus faecalis and Pseudomonas aeruginosa monospecies biofilms on macrophages. METHODOLOGY: In vitro biofilms of E. faecalis and P. aeruginosa were grown (7 days) in aerobic and anaerobic conditions. The biofilm matrix components: exopolysaccharides (EPS) and extracellular DNA (eDNA) were extracted and quantified. The inflammatory potential of EPS and eDNA was assessed on macrophage cell lines (RAW 267.4) using nitric oxide (NO), and enzyme-linked immunosorbent assay for tumour necrosis factor (TNF-α) and interleukin-6 (IL-6) expressions. LPS from P. aeruginosa and planktonic bacteria were positive controls. One-way analysis of variance and the Tukey post hoc test were used for statistical analysis. RESULTS: Extracted EPS from both biofilm strains was associated with higher levels than eDNA in both growth conditions (P < 0.05). The biofilm components had less inflammatory potential compared to planktonic bacteria and LPS. EPS produced higher levels of inflammatory response compared to eDNA for both strains (P < 0.05). IL-6 and TNF-α, and NO expression showed no difference for E. faecalis EPS (P ≥ 0.05). In contrast, P. aeruginosa EPS and eDNA had significant levels of IL-6 compared to TNF-α and NO (P < 0.05). CONCLUSIONS: Monospecies biofilm matrix EPS and eDNA from the bacterial strains tested had the ability to induce a low-grade inflammatory response when compared to planktonic bacteria and LPS. This study highlights the potential of biofilm matrix/components, devoid of bacteria to induce low-grade chronic inflammation.

Biomechanical studies on the effect of iatrogenic dentin removal on vertical root fractures.

INTRODUCTION: The aim of this study was to understand the mechanism by which iatrogenic root dentin removal influences radicular stress distribution and subsequently affects the resistance to vertical root fractures (VRF) in endodontically treated teeth. MATERIALS AND METHODS: The experiments were conducted in two phases. Phase 1: freshly extracted premolar teeth maintained in phosphate-buffered saline were instrumented to simulate three different degrees of dentin removal, designated as low, medium, and extreme groups. Micro-Ct analyzes were performed to quantitatively determine: (a) the amount of dentin removed, (b) the remaining dentin volume, and (c) the moment of inertia of root dentin. The specimens were then subjected to thermomechanical cycling and continuous loading to determine (a) the mechanical load to fracture and (b) dentin microcracking (fractography) using scanning electron microscopy. Phase 2: Finite element analysis was used to evaluate the influence of dentin removal on the stress distribution pattern in root dentin. The data obtained were analyzed using one-way ANOVA and Tukey's post hoc test (P < 0.05). RESULTS: Phase 1: A significantly greater volume of dentin was removed from teeth in extreme group when compared to low group (P < 0.01). The mechanical analysis showed that the load to fracture was significantly lower in teeth from extreme group (P < 0.05). A linear relationship was observed between the moment of inertia and load to fracture in all experimental groups (R2 = 0.52). Fractography showed that most microcracks were initiated from the root canal walls in extreme group. Phase 2: The numerical analysis showed that the radicular stress distribution increased apically and buccolingually with greater degree of root canal dentin removal. CONCLUSIONS: The combined experimental/numerical analyses highlighted the influence of remaining root dentin volume on the radicular bending resistance, stress distribution pattern, and subsequent propensity to VRF.

Microtissue engineering root canal dentine with crosslinked biopolymeric nanoparticles for mechanical stabilization.

AIM: To evaluate the functional strain distribution pattern in root dentine following canal preparation and root canal surface engineering with crosslinked biopolymeric nanoparticles using digital moiré interferometry (DMI). METHODOLOGY: Root dentine specimens were prepared, grating material replicated and tested for 10-50 N, compressive loads in a customized high-resolution, whole-field moiré interferometry set-up. Digital moiré fringes were acquired to determine the strain distribution pattern at specific regions of interest before and after canal enlargement, and dentine surface engineering with a chitosan nanoparticle crosslinker solution. Fringe patterns were acquired, and strain distribution pattern in the direction perpendicular to dentinal tubules (U-field) and parallel to dentinal tubules (V-field) was analysed with custom digital image-processing software. Data were analysed with a statistical method on trend analysis at 0.05 significant level. RESULTS: Distinct deformation patterns perpendicular to the dentinal tubules were observed in root dentine. Root canal dentine removal following instrumentation resulted in an increase in strain distribution, which increased with an increase in applied loads (P < 0.01). The root canal dentine engineered with crosslinked nanoparticles demonstrated a conspicuous decrease in previously increased strain distribution in both coronal and apical root dentine (P < 0.01). A significant increase in tensile strain in root dentine was observed subsequent to instrumentation in the direction parallel to dentinal tubules (P < 0.01). There was a significant reduction in the tensile strain formed at the apical region of the instrumented root dentine following crosslinked nanoparticle treatment (P < 0.05). CONCLUSIONS: This study highlighted the potential of root canal dentine microtissue engineering with crosslinked chitosan nanoparticle to improve radicular strain distribution patterns in instrumented canals.

Photodynamically crosslinked and chitosan-incorporated dentin collagen.

A lingering concern with restored root-filled teeth is the loss of structural integrity of the dentin and dentin-sealer interface over time. We hypothesized that crosslinking of dentin collagen with simultaneous incorporation of a biopolymer into collagen matrix would improve its structural stability. This study aimed to investigate the effects of combining chemical/photodynamic crosslinking of dentin collagen with the incorporation of carboxymethyl-chitosan (CMCS) on the resistance to enzymatic degradation and mechanical properties of dentin collagen. Ninety-six demineralized dentin collagen specimens (human, n = 72; and bovine, n = 24) were prepared and crosslinked chemically/ photodynamically, with/without CMCS. Glutaraldehyde and carbodiimides were used for chemical crosslinking, while rose Bengal activated with a non-coherent light (540 nm) at 20 J/cm(2) was applied for photodynamic crosslinking. The crosslinked human dentin collagen was subjected to chemical characterization, 7 days enzymatic degradation, and transmission electron microscopy (TEM), while the bovine dentin collagen was used for tensile-testing. Crosslinked collagen showed significantly higher resistance to enzymatic degradation (p < 0.01), stable ultrastructure, and increased tensile strength (p < 0.05). Crosslinking CMCS with collagen matrix as observed in the TEM further improved the mechanical properties of dentin collagen (p < 0.01). This study highlighted the possibility of improving the resistance and toughness of dentin collagen by chemically/photodynamically crosslinking collagen matrix with CMCS.

Bioactivity of novel carboxymethyl chitosan scaffold incorporating MTA in a tooth model.

AIM: To characterise the bioactivity of a novel carboxymethyl chitosan (CMCS) scaffold with and without incorporating mineral trioxide aggregate (MTA) in a tooth model. METHODOLOGY: Cross-linked CMCS scaffold (CaC) and MTA-coated CaC (CaMT) scaffold were prepared by freeze-drying. The bioactivity of the scaffolds was tested in vitro in four different mineralisation solutions (bulk system) and ex vivo in simulated body fluid (SBF) in the tooth model. After mineralisation, the mineral deposits on the scaffolds were analysed using scanning electron microscopy, energy dispersive X-ray, and inductively coupled plasma mass spectroscopy. All data were statistically analysed using the two-sample t-test (P < 0.05). RESULTS: Hydroxyapatite (HAP) deposition was observed on CaC and CaMT scaffolds after 1 week of mineralisation in the tooth model and in the bulk system. The deposition was significantly higher (P < 0.05) on CaMT scaffold than that on CaC scaffold. The amount of HAP formed in the tooth model was significantly lower (P < 0.05) than that in the bulk solution. CONCLUSIONS: The CMCS scaffolds are bioactive and capable of biomineralisation by forming HAP within a tooth model ex vivo. The bioactivity of the CMCS scaffold can be enhanced by incorporating MTA.

Influence of bacterial growth modes on the susceptibility to light-activated disinfection.

AIM: To evaluate the efficacy of light-activated disinfection (LAD) using methylene blue (MB) and a non-coherent light source on gram-positive and gram-negative bacteria in different growth modes. The influence of different photosensitizer (PS) formulations in the MB-mediated LAD of biofilms was also evaluated. METHODOLOGY: Light-activated disinfection using MB was tested on Enterococcus faecalis in a planktonic suspension, coaggregated suspension and mono-species biofilms and on Pseudomonas aeruginosa in a planktonic suspension and mono-species biofilms. Further, the difference in susceptibility of E. faecalis and P. aeruginosa biofilms to LAD with modified PS formulations was assessed by conventional culturing methods and confocal laser scanning microscopy (CLSM). RESULTS: Higher energy dose was required for LAD of bacteria in a coaggregated suspension and in biofilm compared to their planktonic counterparts. Biofilm mode of growth offered the greatest resistance to LAD in both the tested strains of pathogens (P<0.001). Gram-positive E. faecalis was more susceptible to LAD than the gram-negative P. aeruginosa, and the use of modified PS formulations was found to enhance the efficacy of LAD to destroy the biofilm (P<0.001). CONCLUSIONS: Bacterial growth modes play a vital role in influencing the susceptibility to LAD in a dose-dependent manner. The nature of the PS formulation influences the susceptibility of biofilms to LAD.

Characterizing bubble dynamics created by high-intensity focused ultrasound for the delivery of antibacterial nanoparticles into a dental hard tissue.

Hig hintensity focused ultrasound (HIFU) has been applied for drug delivery in various disease conditions. Delivery of antibacterial-nanoparticles into dental hard tissues may open up new avenues in the treatment of dental infections. However, the basic mechanism of bubble dynamics, its characterization, and working parameters for effective delivery of nanoparticles, warrants further understanding. This study was conducted to highlight the basic concept of HIFU and the associated bubble dynamics for the delivery of nanoparticles. Characterization experiments to deliver micro-scale particles into simulated tubular channels, activity of ultrasonic bubbles, and pressure measurement inside the HIFU system were conducted. Subsequently, experiments were carried out to test the ability of HIFU to deliver nanoparticles into human dentine using field emission scanning electron micrographs (FESEM) and elemental dispersive X-ray analysis (EDX). The characterization experiments showed that the bubbles collapsing at the opening of tubular channels were able to propel particles along their whole length. The pressure measured showed sufficient negative and positive pressure suggesting that the bubble grew to a certain size before collapsing, thus enabling the particles to be pushed. The FESEM and EDX analysis highlighted the ability of HIFU to deliver nanoparticles deep within the dentinal tubules. This study highlighted the characteristics and the mechanism involved of the bubbles generated by the HIFU and their capability to deliver nanoparticles.

Light activated disinfection: an alternative endodontic disinfection strategy.

BACKGROUND: An improved light activated disinfection technique utilizing a specific photosensitizer formulation, liquid optical-conduit, oxygen-carrier and light energy of appropriate wavelength has been introduced recently. This study tested the efficacy of this improved light activated disinfection on ex vivo biofilms of Enterococcus faecalis at two different stages of maturation. METHODS: Eighty-five tooth sections were prepared and endodontic biofilm of E. faecalis were grown within the root canal. In stage 1, conventional light activated disinfection (LAD), chemical disinfectant (sodium hypochlorite) and improved LAD were tested on four-day-old (immature) biofilms. In stage 2, conventional LAD, improved LAD and chemomechanical disinfection (alone and in combination with improved LAD) were tested on four-week-old (mature) biofilms. RESULTS: Sodium hypochlorite and improved LAD showed the ability to significantly inactivate bacteria in four-day-old biofilms when compared to the control and LAD (p < 0.05). Inactivation of bacteria from deeper dentine was higher in improved LAD than sodium hypochlorite. In four-week-old biofilms, a combination of chemomechanical disinfection and improved LAD produced significant bacterial killing compared to either chemomechanical disinfection or improved LAD alone. CONCLUSIONS: This study highlighted the potential of improved LAD to kill bacteria within dentinal tubules. In combination with chemomechanical preparation, the improved LAD significantly inactivated four-week-old biofilm bacteria.

Hydromechanics in dentine: role of dentinal tubules and hydrostatic pressure on mechanical stress-strain distribution.

OBJECTIVES: This investigation is to understand the role of free water in the dentinal tubules on the mechanical integrity of bulk dentine. METHODS: Three different experiments were conducted in this study. In experiment 1, three-dimensional models of dentine with gradient elastic modulus, homogenous elastic modulus, and with and without hydrostatic pressure were simulated using the finite element method. Static compressive loads of 15, 50 and 100 N were applied and the distribution of the principal stresses, von Mises stresses, and strains in loading direction were determined. In experiment 2, experimental compression testing of fully hydrated and partially dehydrated dentine (21 degrees C for 72 h) was conducted using a Universal testing machine. In experiment 3, Fourier transform infrared spectroscopic analysis of hydrated and partially dehydrated dentine was carried out. RESULTS: The finite element analysis revealed that the dentine model with simulated hydrostatic pressure displayed residual tensile stresses and strains in the inner region adjacent to the root canal. When external compressive loads were applied to the model, the residual stresses and strains counteracted the applied loads. Similarly the hydrated specimens subjected to experimental compression loads showed greater toughness when compared to the partially dehydrated specimens. The stress at fracture was significantly higher in partially dehydrated specimens (p=0.014), while the strain at fracture was significantly higher in hydrated dentine specimens (p=0.037). SIGNIFICANCE: These experiments highlighted the distinct role of free water in the dentinal tubules and hydrostatic pressure on the stress-strain distribution within the bulk dentine.

Enterococcus faecalis-mediated biomineralized biofilm formation on root canal dentine in vitro.

Enterococcus faecalis is the most predominant bacteria in teeth with failed root canal therapy and is found to survive harsh conditions prevailing in the root canals of endodontically treated teeth. This study aims to investigate the interaction between E. faecalis and root canal dentine substrate. Towards this end, tooth specimens were prepared and divided into two groups. The tooth specimens in group 1 were incubated with E. faecalis for periods of 2-, 4-, and 6-week intervals and the chemical composition of the biofilm was determined using X-ray diffraction and Fourier transform infrared (FTIR) spectroscopy. The tooth specimens in group 2 were incubated with E. faecalis for a period of 6 weeks and the topography and ultrastructure of the biofilm were examined using scanning electron microscopy (SEM), light microscopy, and laser confocal scanning microscopy. The sediments formed from the bacterial interaction on the dentine (in group 1) were also examined by SEM and FTIR. These experiments highlighted different stages in the interaction of E. faecalis with root canal dentine. Further, a bacterial-induced apatite reprecipitation on mature biofilm was also observed. This ability of E. faecalis to form such calcified biofilm on root canal dentine may be a factor that contributes to their persistence after endodontic treatment.

Digital moiré interferometric investigations on the deformation gradients of enamel and dentine: an insight into non-carious cervical lesions.

OBJECTIVES: The objective of this study was to evaluate the biomechanical basis of non-carious cervical lesions by examining the patterns of deformation (strain) in the enamel and dentine. METHODS: The digital moiré interferometry is optics based non-destructive, whole-field experimental technique that provides whole-field strain information. Diffraction gratings (with a frequency of 1200 lines/mm) were transferred onto sagittal sections of human teeth, which were subsequently loaded compressively for loads ranging from 10 to 200 N at the incisal edge of the tooth. The acquired digital moiré fringe patterns were used to determine the in-plane deformation pattern in the enamel and the dentine in the direction parallel to the long axis (axial direction) and in the direction perpendicular to the long axis (lateral direction) of the tooth. RESULTS: It is observed that the enamel displayed marked strain gradients in the lateral direction, while the coronal dentine experienced marked strain gradients in the axial directions during compression. With the increase in applied loads, the strains in the enamel increased at the cervical edge (above the cemento-enamel junction) on the facial side, while the strains in the dentine increased below the cemento-enamel junction on the facial side. CONCLUSION: The enamel and dentine displayed unique in-plane deformation patterns in the axial and the lateral directions of the tooth. These experiments support the hypothesis that occlusal loading will contribute to cervical loss of dental hard tissues.