Evaluating the effect of using anti-stress balls as a distraction technique in reducing pain during inferior alveolar nerve block injection: a randomized clinical trial.

OBJECTIVE: The aim of this study is to evaluate the effect of using anti-stress balls in reducing patients' pain during injection of the inferior alveolar nerve block (IANB). MATERIALS AND METHODS: In this randomized clinical trial, 32 individuals were divided into two groups. The conventional method of anesthesia injection was performed using IANB conventional injection technique. During the injection, individuals in the anti-stress ball group were asked to use the anti-stress ball as a distraction technique. For the control group, no supersede methods were used for pain control. Finally, both groups were asked to record their pain utilizing the numerical rating scale (NRS). The participants' vital signs were monitored before and after injection. Kolmogorov-Smirnov test, independent T-test, and Fisher's exact chi-square test were performed for statistical analysis (α = 0.05). RESULTS: Sixteen females and 16 males in the age range of 40 to 20 years old participated in this study. The mean pain score in the anti-stress ball group was significantly lower (p < 0.001). In both sexes, the pain score in the anti-stress ball group was significantly lower (males p < 0.001 and females p = 0.001). In addition, in all age ranges, the pain score in the control group was higher except for the above 35 years old participants (p = 0.078). Moreover, there were no significant differences in individuals' vital signs (p > 0.05). CONCLUSION AND CLINICAL RELEVANCE: Utilizing an anti-stress ball reduces patients' pain significantly during IANB in both sexes and individuals who are below 35 years without changing vital signs. CLINICAL REGISTRATION NUMBER: IRCT20220815055704N1.

Micro-computerized tomographic analysis of premolars restored with oval and circular posts.

OBJECTIVES: The purpose of the study was to evaluate by micro-computerized tomography (microCT) areas and volumes of post, cement, and voids/bubbles in the post space of oval-shaped premolars restored either with oval or circular posts. MATERIALS AND METHODS: Twelve extracted premolars were divided into two groups according to the drill-fiber post system used: (1) GC Fiber Post Drill + circular post GC Fiber Post; (2) Ellipson tip(TM) + oval post Ellipson post(TM). Each tooth was scanned using microCT, and areas and volumes of canal, post space, post, cement, and voids at coronal, medium, and apical level were calculated by using a three-dimensional visualization software. Two-way analyses of variance and Tukey tests were used for statistical analysis (p < 0.05). RESULTS: The area of voids was significantly greater at apical level of oval posts, but no difference was found between the levels among the groups. Regardless of post shape, the volume of voids and of cement was significantly higher at the coronal level. With oval posts, the total volume of cement was twice as much as with circular posts, and the difference was statistically significant. CONCLUSIONS: Our results indicate that the volume of voids does not depend on post shape. Moreover, the microCT analysis demonstrated that the volume of cement was greater with oval posts compared to circular posts. CLINICAL RELEVANCE: The microCT analysis provided interesting information on voids distribution and on the ratio between post shape and cement thickness. These results might address the clinician in the reconstruction of teeth with fiber posts.

Porphyromonas gingivalis virulence factors induce toxic effects in SH-SY5Y neuroblastoma cells: GRK5 modulation as a protective strategy.

Periodontitis (PDS) is a chronic inflammatory disease initiated by a dysbiosis of oral pathogenic bacterial species, such as Porphyromonas gingivalis (Pg). These bacteria can penetrate the bloodstream, releasing various endo and exotoxins that fuel the infection, and stimulate toxic inflammation in different compartments, including the brain. However, the specific mechanisms by which PDS/Pg contribute to brain disorders, such as Alzheimer's disease (AD), remain unclear. This study assessed the effects of Pg's virulence factors - lipopolysaccharide (LPS-Pg) and gingipains (gps) K (Kgp) and Rgp - on SH-SY5Y cells. Our results demonstrated that LPS-Pg activated signaling through the Toll-like receptor (TLR)-2/4 induced a significant downregulation of G protein-coupled receptor kinase 5 (GRK5). Additionally, LPS-Pg stimulation resulted in a robust increase in Tau phosphorylation (pTau) and p53 levels, while causing a marked reduction in Bcl2 and increased cell death compared to unstimulated cells (Ns). LPS-Pg also elevated inducible nitric oxide synthase (iNOS) expression, leading to oxidative damage. In cells overexpressing GRK5 via Adenovirus, LPS-Pg failed to increase iNOS and pTau levels compared to GFP control cells. High GRK5 levels also prevented the nuclear accumulation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB). Furthermore, the overexpression of a GRK5 mutant form lacking the nuclear localization signal (ΔNLS) nearly abolished LPS-Pg induced p53 and iNOS upregulation. Finally, we tested whether Kgp and Rgp mediated similar effects and our data showed that both gps caused a marked downregulation of GRK5 leading to increased p53 and pTau levels. In conclusion, this study provides further insight into the toxic effects elicited by Pg in cells and suggests that preventing GRK5 deficiency may be a valid strategy to mitigate Pg-induced toxic effects (i.e. cell death, oxidative damage, and Tau hyperphosphorylation) in SH-SY5Y cells, which are typical molecular hallmarks of neurodegenerative disorders.

Unlocking the Potential of Dental-Derived Mesenchymal Stem Cells in Regenerative Medicine.

Over the past few decades, life expectancy has been increasing in several countries [...].

Chemical, structural and cytotoxicity characterisation of experimental fluoride-doped calcium phosphates as promising remineralising materials for dental applications.

OBJECTIVES: This study aimed at evaluating the cytotoxicity, chemical and structural properties of experimental fluoride-doped calcium-phosphates as potential remineralising materials for dental applications. METHODS: Experimental calcium phosphates were formulated using ß-tricalcium phosphate, monocalcium phosphate monohydrate, calcium hydroxide, and different concentrations of calcium/sodium fluoride salts [(5 wt%: VSG5F), (10 wt%: VSG10F), (20 wt%: VSG20F)]. A fluoride-free calcium phosphate (VSG) was used as control. Each tested material was immersed in simulated body fluid (SBF), (24 h, 15 and 30 days) to assess their ability to crystallise into apatite-like. Cumulative fluoride release was assayed up to 45 days. Moreover, each powder was placed into a medium containing human dental pulp stem cells (200 mg/mL) and their cytotoxicity was analysed using the 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide (MTT) assay (24 h, 48 h and 72 h incubation). These latter results were statistically analysed by ANOVA and Tukey's test (α = 0.05). RESULTS: All the experimental VSG-F materials produced fluoride-containing apatite-like crystals after SBF immersion. VSG20F presented prolonged release of fluoride ions into the storage media (45d). VSG, VSG10F and VSG20F showed a significant cytotoxicity at dilution of 1:1, while at 1:5, only VSG and VSG20F demonstrated a reduction in cell viability. At lower dilutions (1:10, 1:50 and 1:100) all specimens showed no significant toxicity to hDPSCs, but an increase in cell proliferation. SIGNIFICANCE: The experimental fluoride-doped calcium-phosphates are biocompatible and possess a clear ability to evoke fluoride-containing apatite-like crystallisation. Hence, they may be promising remineralising materials for dental applications.

Fracture Strength and Failure Modes of Endodontically Treated Premolars Restored with Compact and Hollow Composite Posts Subjected to Cyclic Fatigue.

Physical and mechanical properties of continuous carbon or glass fiber reinforced endodontic posts are relevant to increase the retention and resistance of the tooth-restoration system. Hollow posts have been recently designed for delivering the luting cement through the post hole, thus enhancing the post-dentin interface by reducing the risk of air bubbles formation. Methods: Three type of endodontic posts, a carbon fiber hollow post, a glass fiber hollow post and a compact glass fiber post were investigated. Mechanical properties of these posts were assessed through bending tests. Teeth were subjected to fatigue cycling and the strength of restored teeth was detected through static tests. Failure modes were investigated through optical and scanning electron microscopy. Results show that composite posts increase the mechanical stability by more than 100% compared to premolars restored with particulate composite. Carbon fiber posts retain the highest strength (1467 N ± 304 N) among the investigated post and core restoration, but an unfavorable type of fracture has been observed, preventing the tooth re-treatment. Instead, more compliant posts (i.e., glass fiber reinforced composite, providing a strength of 1336 N ± 221 N), show a favorable mode of fracture that allows the re-treatment of teeth in the case that failure occurs. Glass fiber hollow posts show a good trade-off between strength and a favorable type of fracture.

Vital Pulp Therapy of Permanent Teeth with Reversible or Irreversible Pulpitis: An Overview of the Literature.

Vital pulp therapy (VPT) has been recently proposed as an alternative approach even in symptomatic mature permanent teeth with deep caries' lesions, aiming to maintain the pulp vitality over time and/or to avoid non-surgical root canal therapy (NSRCT). However, to date, the diagnosis of reversible or irreversible pulpitis is only based on clinical pain quantity and quality, without precisely reflecting the pulp inflammation status. Therefore, the aim of the present study was to provide an overview based on the current scientific literature to demonstrate the clinical effectiveness of VPT on mature permanent teeth, validating the use of hydraulic calcium silicate-based cements and their role in pain management. VPT may be successfully applied not only in mature permanent teeth diagnosed with reversible pulpitis, but also in permanent dental elements with signs and symptoms of irreversible pulpitis. Hydraulic cements showed favorable outcomes in terms of decrease of pro-inflammatory mediators and of post-operative pain. Pain plays a central role in the chance to perform VPT in mature permanent teeth, since it may be considered as a pre-operative diagnostic criterion as well as a treatment success parameter. In addition, proper assessment of pulp inflammation and choice of appropriate materials are key factors in enhancing VPT success.

Fracture resistance of endodontically-treated maxillary premolars restored with different type of posts and direct composite reconstructions: A systematic review and meta-analysis of in vitro studies.

OBJECTIVE: To conduct a systematic review and meta-analysis to assess if the presence of an endodontic post may increase the fracture resistance of endodontically-treated maxillary premolars directly restored with composite. METHODS: A comprehensive systematic literature search according to the PRISMA statement was conducted to assess in vitro studies including endodontically-treated maxillary premolars, restored with different type of posts supporting direct composite reconstructions. Two groups of meta-analyses were performed using fixed effects model and additional Trial Sequential Analysis (TSA) was carried out. Moreover, risk of bias was conducted and quality of evidence for any performed meta-analysis was assessed using Grading of Recommendations Assessment, Development and Evaluation (GRADE). RESULTS: Twenty-four articles met the inclusion criteria and 13 studies also underwent quantitative evaluation. Fracture resistance of endodontically-treated premolars restored with fiber posts was significantly less than sound teeth (p value: < 0.00001), however endodontically-treated premolars with a fiber post provided an increase in fracture resistance when compared to equivalent teeth restored without post (p value: 0.003). TSA revealed high power of both meta-analyses. Moderate risk of bias was reported in 23/24 studies; only 1 study showed high risk of bias. GRADE system showed moderate strength of evidence due to the presence of a study with high risk of bias and to wide confidence intervals. SIGNIFICANCE: Within the limitation of the present systematic review and meta-analysis, it can be concluded that endodontically-treated maxillary premolars restored with a fiber post and direct composite restoration demonstrated increased fracture resistance when compared to equivalent teeth without post.

An in-vitro study investigating the effect of air-abrasion bioactive glasses on dental adhesion, cytotoxicity and odontogenic gene expression.

OBJECTIVE: To assess the microtensile bond strength (MTBS) and interfacial characteristics of universal adhesives applied on dentine air-abraded using different powders. The analysis includes the cytotoxicity of the powders and their effect on odontogenic gene expression. METHODS: Sound human dentine specimens were air-abraded using bioglass 45S5 (BAG), polycarboxylated zinc-doped bioglass (SEL), alumina (AL) and submitted to SEM analysis. Resin composite was bonded to air-abraded or smear layer-covered dentine (SML) using an experimental (EXP) or a commercial adhesive (ABU) in etch&rinse (ER) or self-etch (SE) modes. Specimens were stored in artificial saliva (AS) and subjected to MTBS testing after 24 h and 10 months. Interfacial nanoleakage assessment was accomplished using confocal microscopy. The cytotoxicity of the powders was assessed, also the total RNA was extracted and the expression of odontogenic genes was evaluated through RT-PCR. RESULTS: After prolonged AS storage, specimens in the control (SML) and AL groups showed a significant drop in MTBS (p > 0.05), with degradation evident within the bonding interface. Specimens in BAG or SEL air-abraded dentine groups showed no significant difference, with resin-dentine interfaces devoid of important degradation. The metabolic activity of pulp stem cells was not affected by the tested powders. SEL and BAG had no effect on the expression of odontoblast differentiation markers. However, AL particles interfered with the expression of the odontogenic markers. SIGNIFICANCE: The use of bioactive glass air-abrasion may prevent severe degradation at the resin-dentine interface. Unlike alumina, bioactive glasses do not interfere with the normal metabolic activity of pulp stem cells and their differentiation to odontoblasts.

Poly(ε-caprolactone) reinforced with sol-gel synthesized organic-inorganic hybrid fillers as composite substrates for tissue engineering.

PURPOSE: The importance of polymer-based composite materials to make multifunctional substrates for tissue engineering and the strategies to improve their performances have been stressed in the literature. Bioactive features of sol-gel synthesized poly(ε-caprolactone)/TiO2 or poly(ε-caprolactone)/ZrO2 organic-inorganic hybrid materials are widely documented. Accordingly, the aim of this preliminary research was to develop advanced composite substrates consisting of a poly(ε-caprolactone) matrix reinforced with sol-gel synthesized PCL/TiO2 or PCL/ZrO2 hybrid fillers. METHODS: Micro-computed tomography and atomic force microscopy analyses allowed to study surface topography and roughness. On the other hand, mechanical and biological performances were evaluated by small punch tests and Alamar Blue™ assay, respectively. RESULTS: Micro-computed tomography and atomic force microscopy analyses highlighted the effect of the preparation technique. Results from small punch tests and Alamar Blue™ assay evidenced that PCL reinforced with Ti2 (PCL=12, TiO2=88 wt%) and Zr2 (PCL=12, ZrO2=88 wt%) hybrid fillers provided better mechanical and biological performances. CONCLUSIONS: PCL reinforced with Ti2 (PCL=12, TiO2=88 wt%) and Zr2 (PCL=12, ZrO2=88 wt%) hybrid fillers could be considered as advanced composite substrates for hard tissue engineering.

COVID-19 Outbreak: An Overview on Dentistry.

Coronavirus disease 2019, also called COVID-19, is the latest infectious disease to rapidly develop worldwide [...].

Management of Endodontic and Periodontal Lesions: the Role of Regenerative Dentistry and Biomaterials.

Regenerative dentistry represents a novel interdisciplinary approach involving biomaterials, several molecules and mesenchymal stem cells (MSCs), preferably derived from oral tissues. The pivotal role of MSCs depends on the fact that they can differentiate into different cell lineages and have the strategic role to release bioactive substances that stimulate the renewal and regeneration of damaged tissues. The role of regenerative dentistry is promising in all the branches of dentistry: the most intriguing application is related to the management of endodontic and periodontal defects, overcoming the surgical approach and the implantology as a consequence of a poorly efficient therapeutic plan.

Cytocompatibility and Bioactive Properties of Hydraulic Calcium Silicate-Based Cements (HCSCs) on Stem Cells from Human Exfoliated Deciduous Teeth (SHEDs): A Systematic Review of In Vitro Studies.

The implementation of hydraulic calcium silicate-based endodontic cements (HCSCs) in biologically based endodontic procedures for the primary dentition has been recently investigated, focusing on the biological response of stem cells from human exfoliated deciduous teeth (SHEDs) towards them. The present systematic review aimed to present a qualitative synthesis of the available literature consisting of in vitro assays, which assessed the cytocompatibility and bioactive properties of HCSCs in direct contact with SHEDs. Following the PRISMA statement, an electronic database search was carried out in Medline, Scopus, Embase, Web of Science, and SciELO on March 31st and updated on November 16th, 2020. In vitro studies evaluating the biological response of SHEDs to the treatment with HCSCs were eligible. Within the term biological response, assays assessing the cytocompatibility (i.e., cell viability, migration, proliferation), cell plasticity or differentiation (i.e., osteo/odontogenic marker expression), and bioactivity or biomineralization (i.e., mineralized nodule formation) were included. A total of seven studies were included after the selection process. The study sample comprised an extensive range of cell viability, migration, proliferation, adhesion, and bioactivity assays regarding the biological response of SHEDs towards five different commercially available HCSCs (MTA, ProRoot MTA, Biodentine, iRoot BP Plus, and Theracal LC). Biodentine, MTA, and iRoot BP Plus showed significant positive results in cytocompatibility and bioactivity assays when cultured with SHEDs. The results from in vitro assays assessing the cytocompatibility and bioactivity of the HCSCs MTA, Biodentine, and iRoot BP Plus towards SHEDs support their use in vital pulp treatment for the primary dentition.

Effect of light curing and dark reaction phases on the thermomechanical properties of a Bis-GMA based dental restorative material.

PURPOSE: The effects of light curing units (LCU) and energy doses on the chemical and physical properties of a dental composite were investigated. METHODS: The effects on the chemical and physical properties of a bisphenol A diglycidylether methacrylate (Bis-GMA) based dental restorative material were evaluated through photospectrometry, differential scanning calorimetry, and mechanical measurements. RESULTS: The light curing conditions associated with direct and indirect restorations were replicated in vitro using optical investigation techniques. A slight attenuation resulted independently of the LCU and a strong attenuation was measured for the cement luting a thick inlay, as well as for the deepest layer of a composite filling increment. Calorimetric measurements indicated that the curing degree is very sensitive to the light energy dose rather than to the LCU. Mechanical testing showed a transient phase during which properties increased. The delay of the composite in reaching adequate properties is strongly dependent on the energy dose. CONCLUSIONS: It is recommended that composites subject to unfavorable light curing conditions undergo a prolonged light curing process.

Small molecule GSK-3 antagonists play a pivotal role in reducing the local inflammatory response, in promoting resident stem cell activation and in improving tissue repairing in regenerative dentistry.

Regenerative dentistry is attracting growing interest in the scientific community, mainly because of its translational and promising therapeutic approach. The latest research carried out by the scientific community are aimed at triggering the local cellular response, in order to induce a physiological self-repairing of damaged oral tissues. Such physiological processes mainly involve the activation of local stem cell populations: mesenchymal stem cells, in fact, retain the ability to proliferate and to differentiate towards functional mature elements, thus leading towards healing of damaged tissues. Glycogen Synthase Kinase-3 (GSK-3) is a key-regulator of the Wnt/ß-catenin pathway; it phosphorylates ß-catenin, that then is degraded in the cytosol. The activation of such signalling, mediated by Wnt ligand/receptor association, inhibits GSK-3, leading to translocation of ß-catenin to the nucleus and to gene transcription. Selective inhibitors of GSK-3 have been linked to the activity of Wnt signalling and to the regeneration of injured tissues, including complex dental and oral structures. Small Molecule GSK-3 Antagonists are the most interesting class of molecules acting with a "Bystander effect": reducing local inflammation and local bone resorption and triggering the activity and differentiation of resident "sleeping" MSCs. The aim of this narrative topical review is to describe the current knowledge on the role of small molecule GSK-3 antagonists in regenerative dentistry, with strategic insights towards the translational applications in nanomaterials in dentistry and in dental repairing.

Commitment of Oral-Derived Stem Cells in Dental and Maxillofacial Applications.

Tissue engineering is based on the interaction between stem cells, biomaterials and factors delivered in biological niches. Oral tissues have been found to be rich in stem cells from different sources: Stem cells from oral cavity are easily harvestable and have shown a great plasticity towards the main lineages, specifically towards bone tissues. Dental pulp stem cells (DPSCs) are the most investigated mesenchymal stem cells (MSCs) from dental tissues, however, the oral cavity hosts several other stem cell lineages that have also been reported to be a good alternative in bone tissue engineering. In particular, the newly discovered population of mesenchymal stem cells derived from human periapical inflamed cysts (hPCy-MSCs) have showed very promising properties, including high plasticity toward bone, vascular and neural phenotypes. In this topical review, the authors described the main oral-derived stem cell populations, their most interesting characteristics and their ability towards osteogenic lineage. This review has also investigated the main clinical procedures, reported in the recent literature, involving oral derived-MSCs and biomaterials to get better bone regeneration in dental procedures. The numerous populations of mesenchymal stem cells isolated from oral tissues (DPSCs, SHEDs, PDLSCs, DFSCs, SCAPs, hPCy-MSCs) retain proliferation ability and multipotency; these features are exploited for clinical purposes, including regeneration of injured tissues and local immunomodulation; we reported on the last studies on the proper use of such MSCs within a biological niche and the proper way to storage them for future clinical use.

Mechanical and Thermal Properties of Dental Composites Cured with CAD/CAM Assisted Solid-State Laser.

Over the last three decades, it has been frequently reported that the properties of dental restorative composites cured with argon laser are similar or superior to those achieved with conventional halogen and light emitting diode (LED) curing units. Whereas laser curing is not dependent on the distance between the curing unit and the material, such distance represents a drawback for conventional curing units. However, a widespread clinical application of this kind of laser remains difficult due to cost, heavy weight, and bulky size. Recently, with regard to the radiation in the blue region of the spectrum, powerful solid-state lasers have been commercialized. In the current research, CAD (computer-aided design)/CAM (computer-aided manufacturing) assisted solid-state lasers were employed for curing of different dental restorative composites consisting of micro- and nanoparticle-reinforced materials based on acrylic resins. Commercial LED curing units were used as a control. Temperature rise during the photopolymerisation process and bending properties were measured. By providing similar light energy dose, no significant difference in temperature rise was observed when the two light sources provided similar intensity. In addition, after 7 days since curing, bending properties of composites cured with laser and LED were similar. The results suggested that this kind of laser would be suitable for curing dental composites, and the curing process does not suffer from the tip-to-tooth distance.

Polyacrylic resins regulate transcriptional control of interleukin-6, gp80, and gp130 genes in human gingival fibroblasts.

Studies have failed to identify the molecular mechanisms that regulate the genotoxic and cytotoxic effects of methacrylate resins, which are important in the biocompatibility of dental materials. Interleukin (IL)-6 has a crucial role in the control of acute-phase protein response during inflammation. In humans, the synthesis and release of two major acute-phase proteins, C-reactive protein and serum amyloid A, are regulated by IL-6. This study focused on IL-6 and activation of its receptors gp80 and gp130 in human gingival fibroblasts in order to assess the effects of the commercial acid resins Jet Kit, Unifast, and Duralay on control of inflammation.

Effect of N-acetyl-L-cysteine on ROS production and cell death caused by HEMA in human primary gingival fibroblasts.

Previous investigations have shown that 2-hydroxyethyl methacrylate (HEMA) causes reactive oxygen species (ROS) production, which in turn affects cell survival and cell death. The purpose of this study was to evaluate the effects of the antioxidant N-acetyl-L-cysteine (NAC) on HEMA-induced toxicity in human primary gingival fibroblasts (HGF). HGF were treated with various concentrations of HEMA (0-12 mm) in the absence and presence of NAC (1, 5, and 10 mm). The 3-(4,5 dimethyiazol-2-1)-2-5-diphenyl tetrazolium bromide (MTT) assay was used to evaluate the mitochondrial dehydrogenase activity after HEMA exposure. Viability and cell death were determined by flow cytometry using Annexin V and PI staining. ROS production was detected by the increasing fluorescence of the oxidation-sensitive dye 2',7'-dichlorofluorescein diacetate (DCFH-DA) after HEMA treatment. After a 24h incubation period, HEMA concentrations higher then 10mm caused a decrease of cell viability, mitochondrial activity, and an increase of cell death. HEMA concentrations of 4-12 mm markedly increased ROS levels in a dose-dependent manner. High NAC concentrations (5 and 10 mm) significantly reduced cell death, and restored the mitochondrial activity after a 24 h co-treatment, but 1 mm NAC increased HEMA toxicity (p<0.05). All NAC concentrations significantly reduced ROS levels induced by HEMA after a 2 h exposure (p<0.05), but no such reduction was observed after a 4 h treatment. Furthermore, treatment with 10 mm HEMA and 1 mm NAC for 6h caused an increase in ROS levels compared to 10 mm HEMA alone (p<0.05). In conclusion, our results suggest that high NAC concentrations protect HGF against HEMA cytotoxicity by reducing the induced ROS levels.

A 3D analysis of mechanically stressed dentin-adhesive-composite interfaces using X-ray micro-CT.

Dentin bonding systems (DBS) have been developed in order to bond restorative materials (i.e. composite) to tooth tissues when function and integrity have to be re-established. Adhesion to dentin results from the penetration of DBS into the demineralised substrate constituted by a conditioned collagen network. The long-term stability of a restored tooth is mainly affected by the seal of the restorative material on the dental structures. Although leakage through the dentin-DBS interface has been widely reported, 3D investigation technique and accurate non-destructive measurements of leakage as functions of mechanical cycling have never been provided. To address these issues, the properties of the material interface are analysed using micro-tensile static and dynamic tests, assisted by the finite element modelling and by the X-ray computed micro-tomography. The dual energy absorption technique, with the synchrotron beam light, has been developed to investigate, in a non-destructive manner, the effect of mechanical cycling on leakage of a silver nitrate staining solution at the dentin-DBS interface. The effect of the pulpal roof on the stress distribution in the coronal dentin-DBS-composite interface has been investigated and the level at which the state of stress can be assumed to be uniform within acceptable limits has been defined. The tensile static and dynamic results suggest that the adhesive strength for the multi-step DBS resulted significantly higher than the other investigated DBS. Imaging results indicate that 3D leakage occurs radially at the dentin-adhesive interface through the interface itself rather than through the unconditioned dentin bulk; moreover, the dynamic tensile loading allows a more diffuse staining penetration.