Microstructure and mineral composition of dental enamel of permanent and deciduous teeth

Purpose: This study evaluated and compared in vitro the microstructure and mineral composition of permanent and deciduous teeth's dental enamel. Methods: Sound third molars (n = 12) and second primary molars (n = 12) were selected and randomly assigned to the following groups, according to the analysis method performed (n = 4): Scanning electron microscopy (SEM), X‐Ray diffraction (XRD) and Energy dispersive X‐ray spectrometer (EDS). Qualitative and quantitative comparisons of the dental enamel were done. The microscopic findings were analyzed statistically by a nonparametric test (Kruskal‐Wallis). The measurements of the prisms number and thickness were done in SEM photomicrographs. The relative amounts of calcium (Ca) and phosphorus (P) were determined by EDS investigation. Chemical phases present in both types of teeth were observed by the XRD analysis. Results: The mean thickness measurements observed in the deciduous teeth enamel was 1.14 mm and in the permanent teeth enamel was 2.58 mm. The mean rod head diameter in deciduous teeth was statistically similar to that of permanent teeth enamel, and a slightly decrease from the outer enamel surface to the region next to the enamel‐dentine junction was assessed. The numerical density of enamel rods was higher in the deciduous teeth, mainly near EDJ, that showed statistically significant difference. The percentage of Ca and P was higher in the permanent teeth enamel. Conclusions: The primary enamel structure showed a lower level of Ca and P, thinner thickness and higher numerical density of rods. Microsc. Res. Tech., 2010. © 2009 Wiley‐Liss, Inc.     

Lattice fringe studies of (100) defects in enamel crystallites

Lattice imperfections in the structure of shark (Nataprion brevirostrais) enamel crystallites have been studied by means of lattice-imaging in a high-resolution transmission electron microscope. Line defects exhibiting various lengths (100–2500 Å) and widths (10–30 Å) and demonstrating phase behaviour different from that of the host crystal were observed along the fringes representing (100) lattice planes. When defect sites were subjected to an electron beam, an annealing process was observed in which the difference in phase contrast eventually disappeared and the defect itself became an intricate part of the (100)-type lattice planes. A through-focal series of the defects show a reversal in phase contrast. By examining the contrast behaviour under certain focal conditions and by adopting the Fresnel diffraction mode, the refractive index of the defect regions was determined to be lower than that of the apatite crystal. Present findings suggest that the interlayering structure along the (100) planes is responsible for the line defects observed in enamel apatites.     

Resin-tooth interfacial morphology and sealing ability of one-step self-etch adhesives: microleakage and SEM study

OBJECTIVES: To compare microleakage of three self‐etch adhesives and to analyze enamel surface morphology and interfacial morphology of resin–enamel and resin–dentin interface under scanning electron microscope (SEM). EXPERIMENTAL DESIGN: Study was conducted in 65 extracted human premolars. Class V cavities were prepared in 45 teeth and assigned to three groups (n = 15) according to three self‐etch adhesives (OptiBond All‐in‐One, iBond, and Adper Prompt L‐Pop). After restoration, 10 samples from each group were used to assess microleakage at enamel and dentin margin. Five samples from each group were used for analysis of interfacial morphology at resin–enamel and resin–dentin interface under SEM. Remaining 20 teeth were used to prepare flat enamel buccal surfaces to analyze the difference in surface morphology after treatment with three adhesives (n = 5 each) and 36% phosphoric acid treatment (n = 5). PRINCIPAL OBSERVATIONS: At enamel margin, Prompt L‐Pop depicted least leakage of all the three adhesives and also showed best interfacial adaptation under SEM. At dentin margin, OptiBond All‐in‐One showed significant less leakage than iBond and Prompt L‐Pop. On flat enamel surface, phosphoric acid produced the most retentive etching pattern when compared with the three adhesives. CONCLUSION: Prompt L‐Pop showed the best bonding effectiveness in enamel, whereas OptiBond All‐in‐One performed significantly better in dentin. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.     

High resolution SE-I SEM study of enamel crystal morphology

Until recently high resolution TEM was the only imaging mode capable of probing the atomic lattice structure of crystals composing tooth enamel. Studies designed to determine the polyhedral shape of normal enamel crystals and initiation of carious lesions in enamel crystals were hampered and limited by interpretation of two-dimensional TEM images from thin section and freeze fracture replica specimens lacking depth of field. The newly developed SE-I signal mode for SEM (SE-I/SE-II ratio) can produce images of enamel crystals approaching beam diameter dimensions (0.7–2.0 nm), rivaling the resolution of the TEM technique and generating topographic contrasts for three dimensional imaging at very high magnification (≈?1,000,000 X). Ultrathin chromium (Cr) films generate enriched high resolution SE-I contrasts of enamel crystal surfaces and when imaged using an immersion lens field emission SEM operated at high voltage (20–30 KeV) produce unsurpassed topographic contrasts. Since the grain size of Cr is below the resolution of any SEM and is ultrathin (≈?1 nm), then SE-I images can provide a more accurate representation of enamel crystal structure than TEM methodologies. Our SE-I SEM observations of normal human enamel crystals reveal fractured spicules which contain angled flat surfaces delineated by a prominent 2 nm wide SE-I edge brightness contrast. Although microscopic observations often show crystals which are hexagonal in cross-section, in both SEM and TEM many other growth habits, including rectangular or irregular crystals (30–40 nm in width) which contain “notches,” are also observed. More detailed morphological studies are therefore required to determine the most likely habit planes and their relevance to the function of the enamel crystals. The granular appearing fine structural contrast imposed onto <100> lattice planes of sectioned enamel in TEM micrographs is also resolved with topographic contrasts in SE-I micrographs. These granules probably represent one or both of the enamel protein classes.     

Microhardness and SEM after CO2 laser irradiation or fluoride treatment in human and bovine enamel

Background: It remains uncertain as to whether or not CO₂ laser is able to hinder demineralization of enamel. The possibility to use bovine instead of human teeth on anticariogenic studies with laser has not yet been determined. Purpose: To compare the ability of CO₂ laser and fluoride to inhibit caries‐like lesions in human enamel and to test whether a similar pattern of response would hold for bovine enamel. Study Design: Ninety‐six enamel slabs (2 × 2 × 4 mm) (48 from bovine and 48 from human teeth) were randomly distributed according to surface treatment (n = 12): CO₂ laser, 5% sodium fluoride varnish (FV), 1.23% acidulated phosphate fluoride (APF) gel, or no treatment (control). Specimens were subjected to a 14‐day in vitro cariogenic challenge. Microhardness (SMH) was measured at 30 μm from the surface. For ultrastructural analysis, additional 20 slabs of each substrate (n = 5) received the same treatment described earlier and were analyzed by SEM. Results: ANOVA and Tukey test ascertained that CO₂ laser promoted the least mineral loss (SMH = 252^a). Treatment with FV resulted in the second highest values (207^b), which was followed by APF (172^c). Untreated specimens performed the worst (154^d). SEM showed no qualitative difference between human and bovine teeth. APF and control groups exhibited surfaces covered by the smear layer. A granulate precipitate were verified on FV group and fusion of enamel crystals were observed on lased‐specimens. Conclusions: CO₂ laser may control caries progression more efficiently than fluoride sources and bovine teeth may be a suitable substitute for human teeth in studies of this nature. Microsc. Res. Tech. 73:1030–1035, 2010. © 2010 Wiley‐Liss, Inc.     

The structure of (100) defects in carbonated apatite crystallites: a high resolution electron microscope study

Planar defects parallel to (100) with an approximate [1/400] displacement vector have been identified by high resolution transmission electron microscopy and by micro-electron diffraction in the center of synthetic carbonated apatite crystallites. Similar intergrowths, 0.8-1.5 nm in width, have been observed in dental enamel, dentin and bone apatite crystallites. Four possible structural models of the defect core are proposed to explain these experimental features, and computer-simulated lattice images of the models are compared with the experimental images. Typical defects were consistent with a two-dimensional octacalcium phosphate inclusion, one unit cell thick, embedded in an apatite matrix.     

Specimen preparation technique for high resolution transmission electron microscopy studies on model supported metal catalysts

A new technique is described which can be used for preparing transmission electron microscopy (TEM) specimens suitable for high resolution studies on supported metal catalysts. By conventional silicon processing techniques 200 × 200 μm² Si₃N₄ membranes on Si wafers are produced. These membranes are extremely flat and have a uniform thickness of 13 nm. They can be used as a support in various kinds of thin film deposition. A TiO₂ film, optimally structured with respect to the requirements for high resolution TEM work in TiO₂–metal cluster systems, is deposited on the Si₃N₄ layer. It consists of one monolayer of 10–25 nm TiO₂ crystallites. TiO₂ lattice images show that a line resolution down to 0.19 nm is possible. Examples of TiO₂–Pd and TiO₂–Rh are given using respectively photodeposition and impregnation reduction to produce l.5–4 nm metal clusters.     

Wear characteristics of porcelain denture teeth

Surface wear and frictional behavior of porcelain denture teeth were examined with a single-pass sliding technique. The influence of environment, type of surface (enamel or dentin) and repeated passes on the wear characteristics of the teeth were investigated. The enamel surface of the teeth was more resistant to penetration and to surface damage than the dentin surface was. Damage was more severe for double-pass than for single-pass sliding. The wear characteristics were affected similarly by the liquid environments of water and of human saliva. Quantitative differences in the wear parameters investigated were found between the two brands of porcelain denture teeth.     

Effect of laser preparation on adhesion of a self‐adhesive flowable composite resin to primary teeth

The aim of the study was to evaluate the adhesion of a self‐adhering flowable composite resin to primary tooth enamel and dentin after silicon carbide paper (SiC) and laser pretreatment. Adhesive properties were evaluated as shear bond strength (SBS) and scanning electron microscopic (SEM) characteristics. A total 120 primary canine teeth were randomly divided into two groups to study enamel and dentin. Each group was divided into 6 subgroups (n = 10) according to type of surface preparation (SiC or Er:YAG laser) of enamel or dentin. Three methods were used to build cylinders of restoration on tooth surface: OptiBond All‐In‐One + Premise Flowable composite, OptiBond All‐In‐One + Vertise Flow and Vertise flow. After restoration, samples were tested for SBS and failure mode. Twenty eight samples were examined by SEM. The results of the study showed SBS of Vertise Flow was lower than others in enamel and dentin samples pretreated with SiC and in dentin samples pretreated with laser (P < 0.001). Compared to SiC pretreatment, laser pretreatment led to a significantly higher SBS with Vertise Flow on enamel (P < 0.001). Vertise Flow associated with the adhesive led to a higher SBS in enamel and dentin compared to Vertise Flow alone. Adhesive and mixed failure modes were observed more frequently in Vertise Flow groups. SEM images showed that Vertise Flow led to more irregularities on enamel and more open dentinal tubules after laser ablation compared SiC pretreatment. Microsc. Res. Tech. 79:334–341, 2016. © 2016 Wiley Periodicals, Inc.     

Marginal adaptation of composite resins under two adhesive techniques

In the present research, different adhesive techniques were used to set up fillings with composite resins. After the application of etch and rinse or self etch adhesive technique, marginal adaptation of composite fillings was estimated by the length of margins without gaps, and by the microretention of resin in enamel and dentin. The study material consisted of 40 extracted teeth. Twenty Class V cavities were treated with 35% phosphorous acid and restored after rinsing by Adper Single Bond 2 and Filtek Ultimate‐ASB/FU 3M ESPE composite system. The remaining 20 cavities were restored by Adper Easy One‐AEO/FU 3M ESPE composite system. Marginal adaptation of composite fillings was examined using a scanning electron microscope (SEM). The etch and rinse adhesive technique showed a significantly higher percentage of margin length without gaps (in enamel: 92.5%, in dentin: 57.3%), compared with the self‐etch technique with lower percentage of margin length without gaps, in enamel 70.4% (p < .001), and in dentin‐22.6% (p < .05). In the first technique, microretention was composed of adhesive and hybrid layers as well as resin tugs in interprismatic spaces of enamel, while the dentin microretention was composed of adhesive and hybrid layers with resin tugs in dentin canals. In the second technique, resin tugs were rarely seen and a microgap was dominant along the border of restoration margins. The SEM analysis showed a better marginal adaptation of composite resin to enamel and dentin with better microretention when the etch and rinse adhesive procedure was applied.     

Quantitative analyses of the biomineralization of different hard tissues

The primary crystallites of the different developing hard tissues have an apatite structure. However, they have crystal lattice distortions representing an intermediate state between amorphous and fully crystalline. We have applied energy-filtering transmission electron microscopy in the selected area electron diffraction mode to analyse different stages of crystal formation in dentine, bone, enamel and inorganic apatite mineral. We have obtained quantitative information on the degree of crystal lattice distortion using the paracrystal theory of Hosemann and Bagchi.
We have found that the early formed crystallites of the hard tissues being analysed have a paracrystalline character comparable to biopolymers. However, with maturation, the lattice fluctuations of the crystallites of the hard tissues bone, enamel and dentine decrease to form a typical (para)crystalline character. Also the decrease of the organic proportion in the matrix corresponds to the decrease of the lattice fluctuation of the crystallites in the different hard tissues during maturation.     

Chemical and crystallographic study of remineralized surface on initial carious enamel treated with Galla chinensis

To investigate the morphologic, chemical and crystallographic characters of remineralized surface on initial carious enamel treated with Galla chinensis, scanning electron microscopy equipped with energy dispersive analysis spectroscopy were used, and X‐ray microdiffraction (microzone XRD) was used for the first time to analyze in situ the microzone crystallite of remineralized surface on carious enamel. Bovine sound enamel slabs were demineralized to produce initial carious lesion in vitro. Then, the lesions were exposed to a pH‐cycling regime for 12 days of remineralization. Each daily cycle included 4×1 min applications with one of the three treatments: distilled and deionized water (DDW); 1 g/L NaF; 4 g/L G. chinensis extract (GCE). After the treatments, some rod‐like deposits and many irregular prominences were found on GCE‐treated enamel surface, and the intensities of Ca and P signals showed a tendency to increase; Ca:P ratio was significantly higher than that of DDW‐treated enamel. X‐ray microdiffraction showed hydroxyapatite was still the main component of GCE‐treated enamel, and the crystallinity was increased, the crystal lattice changed gently with decreased lattice parameter a. These results indicated the potential of GCE in promoting the remineralization of initial enamel carious lesions, and supported the previous hypothesis about GCE mechanism. Combined with the anti‐bacteria and demineralization inhibition properties of GCE, the natural G. chinensis may become one more promising agent for caries prevention. SCANNING 31: 236–245, 2009. © 2010 Wiley Periodicals, Inc.     

Stress and strain distribution in demineralized enamel: A micro‐CT based finite element study

Physiological oral mechanical forces may play a role on the progression of enamel carious lesions to cavitation. Thus, the aim of this study was to describe, by 3D finite element analysis, stress, and strain patterns in sound and carious enamel after a simulated occlusal load. Micro‐CT based models were created and meshed with tetrahedral elements (based on an extracted third molar), namely: a sound (ST) and a carious tooth (CT). For the CT, enamel material properties were assigned according to the micro‐CT gray values. Below the threshold corresponding to the enamel lesion (2.5 g/cm³) lower and isotropic elastic modulus was assigned (E = 18 GPa against E₁ = 80 GPa, E₂ = E₃ = 20 GPa for sound enamel). Both models were imported into a FE solver where boundary conditions were assigned and a pressure load (500 MPa) was applied at the occlusal surface. A linear static analysis was performed, considering anisotropy in sound enamel. ST showed a more efficient transfer of maximum principal stress from enamel to the dentin layer, while for the CT, enamel layer was subjected to higher and concentrated loads. Maximum principal strain distributions were seen at the carious enamel surface, especially at the central fossa, correlating to the enamel cavity seen at the original micro‐CT model. It is possible to conclude that demineralized enamel compromises appropriate stress transfer from enamel to dentin, contributing to the odds of fracture and cavitation. Enamel fracture over a dentin lesion may happen as one of the normal pathways to caries progression and may act as a confounding factor during clinical diagnostic decisions. Microsc. Res. Tech. 78:865–872, 2015. © 2015 Wiley Periodicals, Inc.     

Comparison of methods for quantifying dental wear caused by erosion and abrasion

Various methods have been applied to evaluate the effect of erosion and abrasion. So, the aim of this study was to check the applicability of stylus profilometry (SP), surface hardness (SH) and focus‐variation 3D microscopy (FVM) to the analysis of human enamel and dentin subjected to erosion/abrasion. The samples were randomly allocated into four groups (n = 10): G1‐enamel/erosion, G2‐enamel/erosion plus abrasion, G3‐dentin/erosion, and G4‐dentin/erosion plus abrasion. The specimens were selected by their surface hardness, and they were subjected to cycles of demineralization (Coca‐Cola®‐60 s) and remineralization (artificial saliva‐60 min). For groups G2 and G4, the remineralization procedures were followed by toothbrushing (150 strokes). The above cycle was repeated 3×/day during 5 days. The samples were assessed using SH, SP, and FVM. For each substrate, the groups were compared using an unpaired t‐test, and Pearson correlation coefficients were calculated (α = 5%). For enamel, both profilometry technique showed greater surface loss when the erosion and abrasion processes were combined (P <0.05). The correlation analysis did not reveal any relationships among SH, SP, and FVM to G2 and G4. There were significant correlation coefficients (–0.70 and –0.67) for the comparisons between the FVM and SH methods in enamel and dentin, respectively, in G1 and G3. Choosing the ideal technique for the analysis of erosion depends on the type of dental substrate. SP was not sufficiently sensitive to measure the effects on dentin of erosion or erosion/abrasion. However, SP, FVM and SH were adequate for the detection of tissue loss and demineralization in enamel. Microsc. Res. Tech., 2013. © 2012 Wiley Periodicals, Inc.     

Shear bond strengths of two newly marketed self-adhesive resin cements to different substrates: A light and scanning electron microscopy evaluation

The purpose of this in vitro study was to compare the shear bond strengths (SBSs) of two newly marketed self-adhesive resin cements (RCs) to enamel, dentin, and lithium disilicate (LiSi) glass ceramic block. Forty-eight enamel and 48 dentin substrates were obtained from sound human molars. Additionally, 6 × 7 × 5 -mm- sized 24 specimens were produced from LiSi glass ceramic blocks. The tooth specimens were randomly assigned into four groups (n = 12) according to the surface treatments: (1) G-CEM ONE (GCO), (2) G-CEM ONE Adhesive Enhancing Primer (GCO-AEP) + GCO, (3) RelyX Universal (RXU), and (4) Scotchbond Universal Plus (SUP) + RXU. LiSi specimens were randomly divided into two groups (n = 12): (1) G-MultiPrimer (GMP) + GCO and (2) SUP + RXU. Following the RC applications, all specimens were kept in 100% humidity at 37°C for 24 hr and then submitted for SBS testing in a universal testing machine (1 mm/min). Data were analyzed by Welch's, one-way analysis of variance and two independent samples t tests. The nature of failures was examined under a light microscope, and scanning electron microscopy analyses were also performed for interfaces. GCO and RXU showed similar SBS to enamel (p > .05), and the use of adhesives resulted in improved SBS (p < .05). No difference was detected between GCO-AEP + GCO and SUP + RXU. The GCO-AEP + GCO exhibited the highest SBS to dentin (p < .05), followed by GCO ≥ SUP + RXU > RXU (p < .05). There was no significant difference between SBSs of two RCs to LiSi blocks (p > .05). No cohesive failure was determined for the tested groups by light microscope. The use of adhesives prior to the application of self-adhesive RCs improved their bonding to tooth tissues. GCO demonstrated superior SBS to dentin, whereas both self-adhesive RCs generated similar SBS to enamel and LiSi glass ceramic surfaces.     

Fine structure of tooth enamel in the yellowing human teeth: SEM and HRTEM studies

Objective : The aim of this study was to clarify an influence of the fine structure of human tooth enamel to the yellowing teeth. Materials and methods : Sound maxillary first premolars of 15–50‐year‐old females that were extracted for the orthodontic treatment were used as the test samples. The tooth enamel sections of these teeth that prepared by ion polishing were observed by scanning electron microscopy (SEM). Furthermore, the fine structure of substance filling the inter‐rod spaces was analyzed by high resolution transmission electron microscopy (HRTEM). Results : In white tooth, the inter‐rod spaces were observed at the width of about 0.1 μm, while in yellow tooth, the inter‐rod spaces were not clearly observed by SEM. HRTEM observations revealed for the first time that the inter‐rod spaces were filled with fine particles of poorly crystallized hydroxyapatite in the yellow tooth. In yellow tooth, it was considered that the color of the inner dentin was recognized due to the decrease of light scattering by filling the tooth enamel inter‐rod spaces. The generation of particles in the tooth enamel inter‐rod spaces was considered to be caused by the long‐time progression of calcification. Conclusions : These results suggested that the change in fine structure, filling in inter‐rod spaces of tooth enamel, was related to progression of calcification in the inter‐rod spaces with advancing age and one of the factors of yellowness of human tooth. Microsc. Res. Tech. 79:14–22, 2016. © 2015 Wiley Periodicals, Inc.     

TEM纳米尺度石墨标样的制备

透射电镜的长度标尺或公称放大倍数是判断试样中组织细节尺寸的依据,需要应用纳米级长度标样进行校准。本文介绍了一种用石墨制备透射电镜专用纳米尺度标样的方法,对石墨的X射线衍射分析以及对石墨标样的TEM高分辨像和电子衍射分析表明,标样中石墨的高分辨像为（002）晶面的点阵像,其晶面间距为0．342nm,可作为纳米尺度的参照材料。本文对使用纳米尺度标样校正ETEM标尺的基本方法进行了简要讨论。     

High resolution electron microscope study of lattice images in biological apatites.

Lattice (Fourier) images of crystallites in human bone and teeth, and calcified atherosclerotic plaque were studied using high resolution transmission electron microscope techniques. The lattice images observed in the normal and diseased calcified tissue were compared with the images of synthetic hydroxyapatite crystallites.     

A histopathological study of the pulp of dogs’ teeth after induction of experimental pulp inflammation for different periods of time

The purpose of this study was to investigate the reaction of the pulp of dogs’ teeth after insertion of soft carious dentin from freshly extracted human teeth into the buccal cavities for short and longer periods of time. Forty‐seven mature lower and upper teeth were used in this study. On the middle of the buccal side of the teeth, 3 × 5 mm class ν cavities were prepared, soft carious dentin from freshly extracted human teeth was inserted into the floor of the cavity, and those were filled with glass ionomer. The dogs were killed after 7, 14, 28 and 47 days using vital perfusion techniques. Six‐micrometre sections were prepared and blindly evaluated by pathologist. An inflammatory reaction occurred in all of the samples. Moderate to severe inflammation were shown in all periods except in one in the 7‐day period. Insertion of soft carious dentin gathered from freshly extracted human teeth into class ν cavities produced in dogs’ teeth for 7 days is a reproducible process and could be used in study of pulp pathology.     

Does laser etching have an effect on application mode of a universal adhesive?—A microleakage and scanning electron microscopy evaluation

This study aimed to evaluate the microleakage of a universal adhesive's different application modes incorporated with Er,Cr:YSGG laser on Class V resin composite restorations. Sound human molar teeth (n = 30) were used for microleakage evaluations. Specimens with 60 standardized Class V cavities were divided into five groups according to the adhesive modes of universal adhesive, Adhese Universal (n = 12). Group 1‐etch‐and‐rinse mode with phosphoric acid; Group 2‐etch‐and‐rinse mode with Er,Cr:YSGG laser; Group 3‐selective‐etch mode with phosphoric acid; Group 4‐selective‐etch mode with Er,Cr:YSGG laser; Group 5‐self‐etch. After restorations were performed with a resin composite, Tetric N‐Ceram, the specimens were polished and subjected to thermocycling (10,000X). Following immersion in 0.5% basic fuschin for a day, the teeth were sectioned and the degree of microleakage was determined along the tooth‐resin composite interface using a light microscopy(40X). Five specimens from each group were examined by scanning electron microscopy. The Kruskal–Wallis, Siegel Castello, and Wilcoxon tests were used for statistical analyses (α = .05). At the enamel margins, significant differences were obtained among the groups (p < .05). Significantly higher microleakage scores were detected in Group 5 in comparison with Groups 1, 2, and 3. There were no significant differences between different adhesive strategies at the dentin margins (p > .05). While analyzing enamel and dentin microleakage scores, no statistically significant differences were observed in Groups 4 and 5 (p > .05). The laser application time and the adhesive modes of universal adhesives could affect the microleakage at the enamel margins. Different adhesive modes of universal adhesives combined with laser etching had no influence on the microleakage scores of dentin margins.