Three-Dimensional Assessment of Crown Size and Eruption Space for Developing Third Molars: Data Collection Techniques Based on Cone-Beam Computed Tomography (CBCT).

Third molar development and eruption are two related areas of major interest in dental research into the etiology of "wisdom tooth" impaction. Third molars are not only an excellent model for studying dental development but also of fundamental clinical importance because they are very frequently impacted. Because the third molar is located in the distal-most region of the oral cavity, clinical access is relatively challenging. With the increasingly widespread use of cone-beam computed tomography (CBCT) in dentistry, studies and measurements of the third molar and its eruption area have become considerably easier to do. Here we present a novel CBCT-based measurement methodology we developed for our recent investigations that we hope will also be useful for the broader dental research community.

Use of XRD and SEM/EDX to predict age and sex from fire-affected dental remains.

In fire scenarios, the application and accuracy of traditional odontological methods are often limited. Crystalline studies and elemental profiling have been evaluated for their applicability in determining biological profiles (age and sex) from human dentition, particularly fire- and heat-affected dental remains. Thirty-seven teeth were paired according to tooth type and donor age/sex for the analysis of crown and root surfaces pre- and post-incineration using X-ray diffraction (XRD) and scanning electron microscopy (SEM/EDX). In unburned crowns, carbon (C) content showed a positive correlation with age, whereas phosphorus (P) and calcium (Ca) contents showed a negative correlation with age. In unburned roots, C, P and Ca contents also showed significant changes that were opposite of those observed in the crowns. In relation to sex, females exhibited a higher C ratio than males, whereas males showed significantly higher levels of oxygen (O), P and Ca in unburned roots. Incineration resulted in an increase in the crystallite size that correlated with increasing temperature. No differences in hydroxyapatite (HA) crystallite size were found between age groups; however, unburned teeth from females exhibited a larger crystallite size than did those from males. The challenges of using XRD with a 3D sample were overcome to allow analysis of whole teeth in a nondestructive manner. Further studies may be useful in helping predict the temperature of a fire.

New regression formula to estimate the prenatal crown formation time of human deciduous central incisors derived from a Roman Imperial sample (Velia, Salerno, Italy, I-II cent. CE).

The characterization and quantification of human dental enamel microstructure, in both permanent and deciduous teeth, allows us to document crucial growth parameters and to identify stressful events, thus contributing to the reconstruction of the past life history of an individual. Most studies to date have focused on the more accessible post-natal portion of the deciduous dental enamel, even though the analysis of prenatal enamel is pivotal in understanding fetal growth, and reveals information about the mother's health status during pregnancy. This contribution reports new data describing the prenatal enamel development of 18 central deciduous incisors from the Imperial Roman necropolis of Velia (I-II century CE, Salerno, Italy). Histomorphometrical analysis was performed to collect data on prenatal crown formation times, daily secretion rates and enamel extension rates. Results for the Velia sample allowed us to derive a new regression formula, using a robust statistical approach, that describes the average rates of deciduous enamel formation. This can now be used as a reference for pre-industrial populations. The same regression formula, even when daily incremental markings are difficult to visualize, may provide a clue to predicting the proportion of infants born full term and pre-term in an archaeological series.

Morphology and age-related changes in calcospherites of human teeth: an ultrastructural study.

BACKGROUND: Dentine has been examined extensively for age-related physiological changes, but there are limited data on age-related changes at ultrastructural level of dentine. AIM: The present study aimed to examine age-related ultrastructural changes in calcospherites of human dentine under the scanning electron microscope. MATERIALS AND METHODS: Thirty single rooted teeth of North Western adult Indians (18-75 years) were collected from the Department of Oral Health Sciences, PGIMER, Chandigarh. Labiolingual sections were prepared and morphology of calcospherites was studied at different locations (coronal, cervical, midroot and apical) of the tooth. RESULTS: Morphologically, four types (I [small, unfused and discrete], II [partially fused and globular], III [large, completely fused] and IV [structureless]) of calcospherites were seen in the sample. With advancing age, type I calcospherites approached neighbouring crystals and changed their form to type II, which ultimately coalesced and transformed to type III. Results revealed that among different age groups (young, middle and old), calcospherites of only type I, II and III showed statistically significant differences in their shapes using Pearson's Chi-square test. Statistically non-significant differences were obtained in the shapes at different locations of the tooth. CONCLUSIONS: Results showed that calcospherites get fused as age advances. There is a change in the shape of calcospherites based on the location of the tooth.

Transmission electron microscopical study of teenage crown dentin on the nanometer scale.

This is the first transmission electron microscopic study of teenage crown dentin on the nanometer scale. Samples for TEM were prepared by mechanical thinning and chemical polishing that allowed obtaining the electron transparent foils. It was firstly shown that human dentin possesses the layered morphology: the layers are oriented normally to the main axis of a tooth and have the thickness of ~50nm. HA inorganic phase of teenage crown dentin is in the amorphous state. The cellular structure, which was formed from collagen fibers (diameter is ~5nm), are observed near DEJ region in teenage dentin, whereas bioorganic phase of teenage crown dentin near the pulp camera does not contain the collagen fibers. Cracks in dentin thin foils have sharp tips, but big angles of opening (~30°) with plastic zone ahead crack tip. It means that young crown human dentin exhibits ductile or viscous-elastic fracture behavior on the nanometer scale.

Enamelin Directs Crystallite Organization at the Enamel-Dentine Junction.

Enamel is an acellular material formed by the intricate process of amelogenesis. Disruption caused at the initial stages of development, by means of mutations in the ENAM gene encoding the enamelin protein, results in enamel hypoplasia. Little is known about the consequence of ENAM mutation on the enamel structure at a crystallographic level. The aim of this study was to characterize the structure of ENAM-mutated enamel to develop a deeper understanding of the role of enamelin protein during formation with regard to crystal organization. Synchrotron X-ray microdiffraction (SXRD) and scanning electron microscopy (SEM) have been used to measure and correlate enamel crystallography and microstructure in hypoplastic and healthy enamel. Rietveld refinement carried out on 2-dimensional diffraction patterns, collected from the Advanced Photon Source, were used to quantify changes in the preferred orientation (crystallographic texture) within the labial regions of each tooth slice and then correlated with the local microstructure. In general, healthy deciduous incisors displayed a higher degree of crystal organization across the labial surface in comparison with the hypoplastic enamel. ENAM plays the greatest functional role at the enamel-dentine junction (EDJ), as it was the region that exhibited lowest texture relative to unaffected controls. Other areas within the tooth, however, such as the cusp tip, displayed greater organization in line with healthy enamel, suggesting its effects are restricted to the early stages of enamel secretion. Observed clinically, the surface of ENAM-mutated hypoplastic enamel can appear to be normal, yet severe sub-nano and microstructural defects appear beneath the subsurface layer. Quantitative characterization of the crystallographic properties from enamel with known genotype expands the understanding of enamel formation processes and can aid better clinical diagnosis and tailor-made treatment.

In vivo scanning electron microscope assessment of enamel permeability in primary teeth with and without early childhood caries.

BACKGROUND: Over the years, certain primary teeth have been shown to be highly sensitive to dental caries, while others have remained caries-free. It has been hypothesized that this may be attributed to differences in the permeability of the enamel surface. AIM: The aim of the study was to evaluate the hypothesized differences in the permeability of primary tooth enamel in children with and those without Severe Early Childhood Caries (S-ECC) using scanning electron microscopy. MATERIALS AND METHODS: Sixteen children between 3 and 6 years of age were randomly selected and divided into two groups: Group 1, children without S-ECC (n = 8), and Group 2, children with S-ECC (n = 8). In each child, 4 teeth (the maxillary right and left central and lateral incisors) were subjected to evaluation. An impression was made with polyvinylsiloxane impression material, and scanning electron microscopy was used to inspect the negative replicas for droplets. RESULTS: The results indicated higher significance when individual regions (cervical, middle, and incisal thirds) in the two groups were evaluated and compared. Similarly, the overall results showed high statistical significance between S-ECC and non-S-ECC teeth. CONCLUSION: There could be a positive relationship between the permeability of tooth enamel and the development of caries, which needs further research.

Cuspal flexure, depth-of-cure, and bond integrity of bulk-fill composites.

PURPOSE: Evaluate cuspal flexure caused by polymerization shrinkage stress, bond integrity, and depth-of-cure of bulk-fill composites. METHODS: Twenty-eight extracted permanent molars were mounted in stainless steel rings. Slot-shaped Class II mesio-occlusal-distal preparations (four mm deep) were restored with Filtek Supreme Ultra (control; two two-mm increments) and bulk-fill composites (Tetric EvoCeram, Venus, Filtek Bulk Fill). The teeth were digitized using a 3D scanner before and after restoration. Before- and after-restoration scans were aligned, and cuspal flexure was calculated. Bond integrity along occlusal interfaces was assessed by dye penetration and measured after overnight immersion in basic fuchsin dye and cross-sectioning. Depth-of-cure was determined on the cross-sections using Vickers hardness. Statistical analysis was performed with one-way analysis of variance. RESULTS: All composites caused inward cuspal flexure (10.4 to 13.6 µm). No statistical difference in flexure was found among the composites (P=.07). No significant difference in microhardness at any restoration depths was found for any composite (.35

Tubule density and diameter in coronal dentin from primary and permanent human teeth.

This study compared dentinal tubule density and diameter of human primary and permanent teeth at different depths of the coronal dentin. Crowns of eight primary second molars and eight permanent third molars were serially sectioned into three disks of ~0.5 mm thickness (superficial, middle, and deep layers), perpendicular to the long axis. Tubule density and diameter were evaluated in 2,000× and 3,000× magnifications by scanning electron microscopy. Data obtained were subjected to two-way repeated measures ANOVA and Tukey's post hoc test (α = 0.05). Tubule density was greater in primary teeth compared with permanent ones, regardless of depth (primary: 124,329 ± 43,594 mm2; permanent: 45,972 ± 21,098 mm2). In general, the tubule density increased as the dentin depth increased, except to the superficial and middle layers from permanent teeth. Tubule diameter was larger in the dentin layer close to the pulp chamber (superficial: 2.4 ± 0.07 µm; middle: 3.70 ± 0.06 µm; deep: 4.28 ± 0.04 µm). No difference was observed between primary (3.48 ± 0.81 µm) and permanent teeth (3.47 ± 0.73 µm). The tubule diameter increases as the dentin depth increases for primary and permanent teeth; however, the tubule density is higher in primary teeth.

FTIR and SEM analysis of CO2 laser irradiated human enamel.

OBJECTIVES: Considering the enamel chemical structure, especially carbonate band, which has a major role in the caries prevention, the objective of the present study was to assess the chemical alterations on the enamel irradiated with CO(2) laser by means of FTIR spectroscopy and SEM analysis. DESIGN: The enamel surfaces were analysed on a spectrometer for acquisition of the absorption spectrum relative to the chemical composition of the control sample. The irradiation was conducted with a 10.6-µm CO(2) laser (0.55W, 660W/cm(2)). The carbonate absorption band at 1600-1291cm(-1) as well as the water absorption band at 3793-2652cm(-1) was measured in each sample after the irradiation. The water band was measured again 24-h after the irradiation. The band area of each chemical compound was delimited, the background was subtracted, and the area under each band was integrated. Each area was normalized by the phosphate band (1190-702cm(-1)). RESULTS: There was a statistically significant decrease (p<0.05) in the water content after irradiation (control: 0.184±0.04; irradiated: 0.078±0.026), which increased again after rehydration (0.145±0.038). The carbonate/phosphate ratio was measured initially (0.112±0.029) and its reduction after irradiation indicated the carbonate loss (0.088±0.014) (p<0.05). CONCLUSION: The 10.6-µm CO(2) laser irradiation diminishes the carbonate and water contents in the enamel after irradiation.

Evaluation of enamel surfaces after bracket debonding: an in-vivo study with scanning electron microscopy.

INTRODUCTION: The purposes of this in-vivo study were to compare the modes of failure of uncoated and adhesive precoated metal brackets by using the adhesive remnant index, and to assess the quality of the enamel surface after cleanup by using the enamel damage index. METHODS: Twelve Victory brackets (group A) and 12 Victory adhesive precoated brackets (group B) (both, 3M Unitek, Monrovia, Calif) were bonded onto the maxillary second premolars of 12 volunteers. The uncoated brackets were bonded with Transbond XT adhesive resin (3M Unitek). Replicas of the teeth were made before bonding (T0), after bracket removal (T1), and after cleanup (T2). Scanning electron microscope images of all labial enamel surfaces were taken at T0, T1, and T2, and these were evaluated according to the adhesive remnant index and the enamel damage index. RESULTS: Evaluation of the adhesive remnant index scores with the chi-square test showed no statistically significant difference between the groups. Evaluation of the enamel damage index grades with the sign test for paired samples showed a statistically significant difference (P <0.01) between T0 and T2. CONCLUSIONS: Uncoated and precoated brackets exhibited similar debonding patterns. Additionally, the debonding method tested in this study did not restore the original enamel surface, although there was no clinically relevant enamel damage.

Comparison of premolar cuspal deflection in bulk or in incremental composite restoration methods.

OBJECTIVES: This study examined the cuspal deflection of maxillary premolars when either a bulk filling or incremental filling technique was employed using a range of composites with different elastic moduli. METHODS: Four brands of composite materials, Heliomolar (HM, Ivoclar Vivadent, Schaan, Liechtenstein), Heliomolar HB (HH, Ivoclar Vivadent, Schaan, Liechtenstein), Filtec Supreme XT (FS, 3M Dental Product, St Paul, MN, USA), and Renew (RN, Bisco Inc, Schaumburg, IL, USA), as well as three filling techniques, bulk filling, two-layer incremental filling, and three-layer incremental filling methods, were used. One hundred twenty caries-free human premolars were collected and divided into four groups according to the filling material used. Each of these four groups was then subdivided into three groups according to filling method. In group 1, a bulk filling of 0.15 g of each resin was inserted and light-cured with LED light from the occlusal, mesial, and distal surfaces for 60 seconds each. Group 2 was given two horizontal increments, 0.08 g and 0.07 g, with each increment light-cured from the occlusal, mesial, and distal surfaces for 30 seconds each. In group 3, three horizontal increments of 0.05 g were used, each of which was light-cured from the occlusal, mesial, and distal surfaces for 20 seconds each. The cuspal deflection was measured using a customized cuspal deflection measuring machine for 10 minutes after initiating light polymerization. The elastic modulus of each composite resin material was measured using a three-point bending test. Two-way analysis of variance (ANOVA) with a Dunnet test was used to examine the effect of the two variables (curing methods, materials) on the amount of cuspal deflection at the 95% confidence level. In each material, groups 1, 2 and 3 were compared using one-way ANOVA and a Dunnet test at the 95% confidence level. The elastic moduli of HM, HH, FS, and RN were compared using one-way ANOVA and a Tukey test at the 95% confidence level. The relationship between the amount of cuspal deflection in each group and the elastic modulus of the composite was analyzed using a Pearson correlation test. RESULTS: The amount of cuspal deflection in HH was larger than in the other materials (HM, FS, and RN; p<0.05). There was no significant difference between HM, FS, and RN. The amount of cuspal deflection was greatest in group 1, followed in order by groups 2 and 3 (p<0.05). The amount of cuspal deflection was in the following order: group 1≥2≥3 in HM, and 1>2, 3 in HH, FS, and RN. The elastic modulus was HH>RN>FS>HM (p<0.05). There was a positive correlation between the cuspal deflection and the elastic modulus of the composite. CONCLUSIONS: The incremental filling techniques reduced the amount of cuspal deflection in all composite groups with different elastic moduli. The amount of cuspal deflection showed a positive correlation with the elastic modulus of the composite.

Enamel distribution, structure and mechanical alterations in col1-caPPR mice molar.

Enamel mineralisation is a highly controlled process in which the deposition, growth, and maturation of inorganic crystallites are regulated by secreted matrix proteins at the molecular and cellular level. Maxillary and mandibular first molars from the col1-caPPR mutants as well as normal controls aged for 12 weeks were observed by SEM and nanoindentation, respectively. Several types of aberrations in enamel distribution and crystal organisation were encountered in the transgenic molars. Also, the gene alteration resulted in degradation by as much as 23.42% in hardness and 17.56% in the elastic modulus. These data suggested that gene mutation altered the ameloblastic differentiation and movement, resulting in variations of crystal arrangement patterns, aberrations of enamel distribution, and degradation of mechanical behaviour. Furthermore, the col1-caPPR mouse model was determined as useful for studying how the genes modulate the biomineralisation process.

Evaluation of the integrity of dental sealants by optical coherence tomography.

OBJECTIVES: The purpose of the present research was to demonstrate images of failures into the structure of pit and fissure sealants using optical coherence tomography (OCT). METHODS: Five human third molars were selected and the sealant ALPHA SEAL LIGHT/DFL was applied according to the manufacturer instructions. For evaluation of the structure of pit and fissure sealants, twenty OCT images of each tooth were performed before and after application of sealant. A total of 200 images were evaluated. A home built spectral OCT system used was operating in spectral domain (SD-OCT), at 840 nm and a measured spatial resolution of 10 µm. The system is based on the Michelson interferometer set-up and is controlled by the software OCT 800--Complex Square/LabView, that collects data and generates the image. The occlusal surfaces were scanned in a bucco-lingual direction and tomographic images parallel to long axis of tooth were obtained. After the achievement of the images by OCT, the crown of each tooth was sectioned in a occlusogingival direction (buccolingually). Images from each section were obtained and evaluated by an optical microscope. OCT and microscopic images were compared. RESULTS: Representative images showed that OCT image provides the insight into sealant material. It was possible to clearly identify the internal structure of sealant and the subjacent enamel. Sealant and enamel are very well distinguished, and failures at the interior and at surface are well detected. SIGNIFICANCE: OCT could generate images of the features of fissures, bubbles and failures in the adaptation of sealants, opening up possibilities in the future to monitoring sealant application and retention in short and long term.

Effect of adhesives and thermocycling on the shear bond strength of a nano-composite to coronal and root dentin.

OBJECTIVES: To evaluate the effects of "etch & rinse" vs "self-etch" adhesives and thermocycling on the shear bond strength (SBS) of a nano-composite to coronal and root dentin. MATERIALS AND METHODS: FIFTY-SIX EXTRACTED HUMAN MOLARS WERE mounted and ground to expose coronal and root dentin surfaces and were randomly divided into two groups according to adhesive system: SE--a two-step self-etch adhesive (Adper SE Plus, 3M ESPE) and ER-an etch & rinse adhesive (Adper Single Bond 2, 3M ESPE). The adhesives were applied to the coronal and root dentin of the mounted teeth. A nano-composite (Filtek Supreme XT, 3M ESPE) was applied to both dentin surfaces. The bonded specimens were stored in distilled water at 37 degrees C for 24 hours. Half of the bonded specimens were tested for SBS in a universal testing machine without thermocycling. The remaining specimens were thermocycled (500 cycles between 5 degrees C and 55 degrees C) prior to SBS testing. Two specimens from each group were kept for Scanning Electron Microscope evaluations of the adhesive interfaces. Failure modes were determined under a stereomicroscope. The mean SBS value of each group was calculated, and the results were subjected to ANOVA, Duncan test and Paired samples t-test (p = 0.05). RESULTS: Thermocycling did not affect the SBS of coronal dentin for either adhesive. However, it affected the SBS of SE in root dentin (p < 0.05). The two different dentin substrates did not exhibit a significant difference except for higher values in root dentin with the SE group without thermocycling. The greatest number of cohesive failures was observed in root dentin with SE adhesive; however, the failure modes were mainly adhesive for the other groups. CONCLUSION: The SE adhesive exhibited higher SBS values than the ER adhesive in root dentin. Thermocycling did not affect the SBS in coronal dentin for either adhesive but it decreased the SBS of SE in root dentin.

Size, ultrastructure, and microhardness of natal teeth with agenesis of permanent successors.

OBJECTIVE: Natal and neonatal teeth may occur in conjunction with other oral anomalies, including anomalous development of succedaneous teeth. Our purpose was to examine the size, ultrastructure, and microhardness of two natal teeth without permanent successor germs, and compare them with the characteristics of normal primary teeth. DESIGN: The dimensions of two natal mandibular incisors extracted from a healthy 7-year-old child were measured with a digital sliding caliper. The crown dimensions of normal primary mandibular central incisors were established from dental casts of 102 children. Since the natal teeth were nearly completely devoid of enamel, the average enamel thickness determined on a sample of 19 normal primary mandibular incisors was added to the values measured. External surfaces and transverse sections of the natal teeth were examined under a scanning electron microscope. Vickers microhardness of radicular dentine was assessed for the natal teeth and for 11 normal primary mandibular incisors. RESULTS: Only remnants of enamel were present. All crown dimensions corrected for enamel loss were within the normal range. The ultrastructure of hard dental tissues was normal. The radicular dentine hardness of the natal teeth was similar to that of the normal primary mandibular incisors. CONCLUSIONS: The results indicate that the natal teeth were prematurely erupted regular primary mandibular central incisors. In our case, the occurrence of natal teeth associated with agenesis of their primary successors appears to be related to an accelerated or premature pattern of dental development, rather than to superficial positioning of the tooth germs.

Microleakage of orthodontic band cement at the cement-enamel and cement-band interfaces.

INTRODUCTION: Our objective was to determine and compare microleakage patterns of conventional glass ionomer cement (GIC), resin modified GIC (RMGIC), and polyacid-modified composite for band cementation. METHODS: Sixty freshly extracted third molars were randomly divided into 3 groups of 20 teeth each. Microetched molar bands in the 3 groups were cemented to enamel with one of three orthodontic cements: Ketac-Cem (3M ESPE, Gmbh, Seefeld, Germany), Multi-Cure (3M Unitek, Monrovia, Calif), and Transbond Plus (3M Unitek). A dye penetration method was used for microleakage evaluation. Microleakage was determined by a stereomicroscope for the cement-band and cement-enamel interfaces from both the buccal and lingual margins. Statistical analysis was performed with Kruskal-Wallis and Mann-Whitney U tests. RESULTS: The buccal sides had similar microleakage values compared with the lingual sides for the cement-enamel and cement-band interfaces with all cements. Statistical comparisons showed statistically significant differences among the band cements between both interfaces (P <0.001). When the cement systems were compared, conventional GIC showed the highest leakage scores between cement-band (median, 3.50 mm) and cement-enamel (median, 2.88 mm) interfaces. Teeth banded with RMGIC and modified composite showed similar microleakage scores, and both had less leakage (<1 mm) than conventional GIC. CONCLUSIONS: Conventional GIC is associated with more microleakage than RMGIC and modified composite at both the cement-band and cement-enamel interfaces.

Technical note: a new three-dimensional technique for high resolution quantitative recording of perikymata.

The number and spacing of incremental markings at the enamel surface, known as perikymata, are considered important indicators of dental growth patterns, as they provide information on crown formation times and the underlying developmental processes. This study explores the potential of a new three-dimensional technique for the reconstruction of dental growth profiles, using teeth from a medieval child from Abingdon, Oxfordshire. The crowns of three anterior teeth were imaged and analyzed using the Alicona 3D InfiniteFocus imaging microscope. Individual perikyma grooves can be unambiguously identified on a profile of the reconstructed enamel surface and direct distances between successive pairs of perikyma grooves can be calculated from coordinate data. This quantitative approach constitutes a more objective way to record perikymata spacing than current methods.

3D variations in human crown dentin tubule orientation: a phase-contrast microtomography study.

OBJECTIVES: Tubules dominate the microstructure of dentin, and in crowns of human teeth they are surrounded by thick mineralized peritubular cuffs of high stiffness. Here we examine the three-dimensional (3D) arrangement of tubules in relation to enamel on the buccal and lingual aspects of intact premolars and molars. Specifically we investigate the angular orientation of tubules relative to the plane of the junction of dentin with enamel (DEJ) by means of wet, non-destructive and high-resolution phase-contrast (coherent) tomography. METHODS: Enamel capped dentin samples (n=16), cut from the buccal and lingual surfaces of upper and lower premolar and molar teeth, were imaged in water by high-resolution synchrotron-based phase-contrast X-ray radiography. Reconstructed 3D virtual images were co-aligned with respect to the DEJ plane. The average tubule orientation was determined at increasing distances from the DEJ, based on integrated projections onto orthogonal virtual planes. The angle and curl of the tubules were determined every 100 microm to a depth of 1.4mm beneath the DEJ. RESULTS: Most tubules do not extend at right angles from the DEJ. Even when they do, tubules always change their orientations substantially within the first half-millimeter zone beneath the DEJ, both on the buccal and lingual aspects of premolar and molar teeth. Tubules also tend to curl and twist within this zone. Student t-tests indicate that lower teeth seem to have greater tilts in the tubule orientations relative to the DEJ normal with an average angle of 42 degrees (+/-2.0 degrees), whereas upper teeth exhibit a smaller change of orientation, with an average of 32 degrees (+/-2.1 degrees). SIGNIFICANCE: Tubules are a central characteristic of dentin, with important implications on how it is arranged and what the properties are. Knowing about the path that tubules follow is important for various reasons, ranging form improving control over restorative procedures to understanding or simulating the mechanical properties of teeth. At increasing depths of dentin beneath enamel, tubules are significantly tilted relative to the DEJ norm, which may be important to understand clinical challenges such as sensitivity, effectiveness of bonding techniques or prediction of possible paths for bacterial invasion. Our data show dissimilar average tubule angles of upper versus lower teeth with respect to the DEJ which presumably contributes to different shear responses of the tissue under function. The degree to which this may warrant improved restoratives or new adhesive techniques to enhance adhesive restorations merits further investigation.