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.     

Effect of different irrigation protocols on the radicular dentin interface and bond strength with a metacrylate‐based endodontic sealer

This study assessed the influence of different endodontic chemical substances on the adhesion of the Epiphany SE/Resilon system (with and without resinous solvent) to radicular dentin walls, using the push‐out test and scanning electron microscopy (SEM). Forty‐eight root canals of human canines were prepared biomechanically with ProTaper rotary files (crown‐down technique) and the radicular dentin was treated with either 17% EDTA, 2% chlorhexidine gel (CHX) or 2.5% NaOCl (control). The root canals were filled with Resilon cones and Epiphany SE sealer with and without resinous solvent. Six groups of eight canals each had their roots sectioned transversally to obtain 1‐mm thick slices. Data were subjected to statistical analysis by ANOVA and Tukey's tests. The specimens treated with 17% EDTA (1.59 ± 0.91) presented higher bond strength (P < 0.05) than those treated with 2.5% NaOCl (0.93 ± 0.27) and 2% CHX (0.92 ± 0.22). Significantly higher bond strength (P < 0.05) was observed when the Epiphany SE was prepared with (1.37 ± 0.78) than without (0.92 ± 0.33) solvent. Adhesive failures were predominant in all groups. SEM analysis showed greater homogeneity of the filling mass when the solvent was added to the sealer. Treatment of root canal walls with 17% EDTA, and addition of a resinous solvent to Epiphany SE produced the highest adhesion to radicular dentin. Microsc. Res. Tech. 77:446–452, 2014. © 2014 Wiley Periodicals, Inc.     

Oral biofilm formation on the titanium and zirconia substrates

OBJECTIVE: The aim of this randomized crossover investigation was to assess the biofilm formation on two titanium and one zirconia substrates in relation to the topography and surface roughness of the materials. METHODS: Twenty‐four discs specimens for each evaluated material (Machined pure titanium, Cast pure titanium, or Zirconia) were evaluated after oral cavity exposure in six healthy subjects. The study was conducted in 3 phases according to the material evaluated. Each subject was asked to use a removable splint containing 4 disks of the same tested substrate, 2 located in the anterior, and 2 in the posterior region. Participants were asked to use the intraoral splint during 24 h. The total biofilm covering on the discs was evaluated by 1% of neutral red staining. EMV and surface roughness was carried out to correlate with the biofilm found between different substrates. RESULTS: Data showed higher mean roughness values for zirconia (Zc) when compared with titanium specimens (MPT and CPT; P < 0.001). MPT and CPT presented no differences between them (P > 0.05). The mean percentage (%) of covering biofilm on substrates was 84.14 for MPT, 86.22 for CPT and 90.90 for Zc. CONCLUSION: There were no significant differences in the total area of formed biofilm among the tested groups. No correlation was found between surface roughness and the total amount of formed biofilm in the groups. Microsc. Res. Tech., 2013. © 2012 Wiley Periodicals, Inc.     

Error rates in buccal-dental microwear quantification using scanning electron microscopy

Dental microwear, usually analyzed using scanning electron microscopy (SEM) techniques, is a good indicator of the abrasive potential of past human population diets. Scanning electron microscopy secondary electrons provide excellent images of dental enamel relief for characterizing striation density, average length, and orientation. However, methodological standardization is required for interobserver comparisons since semiautomatic counting procedures are still used for micrograph characterization. The analysis of normally distributed variables allows the characterization of small interpopulation differences. However, the interobserver error rates associated with SEM experience and the degree of expertise in measuring striations are critical to population dietary interpretation. The interobserver comparisons made here clearly indicate that the precision of SEM buccal microwear measurements depends heavily on variable definition and the researcher's expertise. Moreover, error rates are not the only concern for dental microwear research. Low error rates do not guarantee that all researchers are measuring the same magnitudes of the variables considered. The results obtained show that researchers tend to maintain high intrapopulation homogeneity and low measurement error rates, whereas significant interobserver differences appear. Such differences are due to a differential interpretation of SEM microwear features and variable definitions that require detailed and precise agreement among researchers. The substitution of semiautomatic with fully automated procedures will completely avoid interobserver error rate differences.     

Effect of final irrigation protocols on microhardness and erosion of root canal dentin

The aim of this study was to evaluate the effect of final irrigation protocols (17% EDTA, BioPure MTAD, SmearClear, and QMiX) on microhardness and erosion of root canal dentin. Fifty roots were sectioned transversely at the cement–enamel junction and each root was sectioned horizontally into 4‐mm‐thick slices. The samples were divided into five groups (n = 10) according to the final irrigation protocol: G1: distilled water (control group); G2: 17% EDTA; G3: BioPure MTAD; G4: SmearClear; and G5: QMiX. The dentin microhardness was then measured with a load of 25 g for 10 s. Initially, the reference microhardness values were obtained for the samples without any etching. The same samples were then submitted to the final irrigation protocols. A new measure was realized and the difference between before and after the procedures was the dentin microhardness reduction. In sequence, the specimens were submitted to SEM analysis to verify the dentinal erosion. The Kruskal Wallis and Dunn tests (α = 5%) were used to compare the results. The dentin microhardness decreased for all final irrigation protocols. There was no significant difference between groups 2, 3, 4, and 5 (P > 0.05), but this groups presented significant dentin microhardness reduction than G1 (P < 0.05). In G2, occurred the highest incidence of dentinal erosion (P < 0.05). 17% EDTA, BioPure MTAD, SmearClear, and QMiX promoted significant dentin microhardness reduction. Dentinal tubules erosion was promoted by 17% EDTA. Microsc. Res. Tech., 76:1079–1083, 2013. © 2013 Wiley Periodicals, Inc.     

Scanning electron microscopy studies of double-layer silicon structures prepar by epitaxy and direct bonding

The electrical properties of multilayer structures obtained by direct bonding of silicon wafers and epitaxial growth have been investigated. The measurements were made by scanning electron microscopy (SEM) in either secondary electron or electron beam-induced current (EBIC) regime, using cross sections of the structures with a p-n junction formed in the subsurface region of the active layer. The measurements of defect recombination activity were made using Schottky diodes formed on the active layer surfaces. Parasitic p-n junctions in some samples under a small direct voltage have been observed and the reason for the appearance of such parasitic junctions has been established. Two types of defects with different distribution densities and amplitudes of EBIC contrast have been detected.     

Developing 3D SEM in a broad biological context

When electron microscopy (EM) was introduced in the 1930s it gave scientists their first look into the nanoworld of cells. Over the last 80 years EM has vastly increased our understanding of the complex cellular structures that underlie the diverse functions that cells need to maintain life. One drawback that has been difficult to overcome was the inherent lack of volume information, mainly due to the limit on the thickness of sections that could be viewed in a transmission electron microscope (TEM). For many years scientists struggled to achieve three‐dimensional (3D) EM using serial section reconstructions, TEM tomography, and scanning EM (SEM) techniques such as freeze‐fracture. Although each technique yielded some special information, they required a significant amount of time and specialist expertise to obtain even a very small 3D EM dataset. Almost 20 years ago scientists began to exploit SEMs to image blocks of embedded tissues and perform serial sectioning of these tissues inside the SEM chamber. Using first focused ion beams (FIB) and subsequently robotic ultramicrotomes (serial block‐face, SBF‐SEM) microscopists were able to collect large volumes of 3D EM information at resolutions that could address many important biological questions, and do so in an efficient manner. We present here some examples of 3D EM taken from the many diverse specimens that have been imaged in our core facility. We propose that the next major step forward will be to efficiently correlate functional information obtained using light microscopy (LM) with 3D EM datasets to more completely investigate the important links between cell structures and their functions.     

Effects of chemical agents on physical properties and structure of primary pulp chamber dentin

This study evaluated the effects of chemical agents on the physical properties and structure of primary pulp chamber dentin using surface roughness, microhardness tests, and scanning electron microscopy (SEM). Twenty‐five primary teeth were sectioned exposing the pulp chamber and were divided into five groups (n = 5): NT, no treatment; SH1, 1% sodium hypochlorite (NaOCl); SH1U, 1% NaOCl + Endo‐PTC^®; SH1E, 1% NaOCl + 17% EDTA; and E, 17% EDTA. After dentin treatment, the specimens were submitted to roughness, microhardness testing, and SEM analysis. Roughness and microhardness data were submitted to one‐way ANOVA and Tukey's test (P < 0.05). The SH1E group showed the highest roughness, followed by the E group (P < 0.05) when compared with the NT, SH1, and SH1U groups. Microhardness values of SH1 and SH1U showed no significant difference as compared to the NT (control) group (P > 0.05). Microhardness values could not be obtained in the EDTA groups (SH1E and E). The presence of intertubular dentin with opened dentin tubules was observed in the NT, SH1, and SH1U groups. SH1E showed eroded and disorganized dentin with few opened tubules and the intertubular/peritubular dentin was partially removed. Considering the physical and structural approaches and the chemical agents studied, it can be concluded that NaOCl and NaOCl associated with Endo‐PTC^® were the agents that promoted the smallest changes in surface roughness, microhardness, and structure of the pulp chamber dentin of primary teeth. Microsc. Res. Tech. 77:52–56, 2014. © 2013 Wiley Periodicals, Inc.     

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.     

Human chromatid ultrastructure: new observations with scanning and transmission electron microscopy

Two new observations have been made on human chromatid/chromosome ultrastructure using both scanning and transmission electron microscopy (SEM, TEM). A bipartite, apparently half-chromatid-like structure was observed in whole human chromosomes studied with SEM and in longitudinally sectioned chromosomes analyzed with TEM. In addition, we also observed a zipper-like configuration as the parallel sister chromatids separated likely due to the supercoiled structure of the chromosome and chromatid. It is possible that either or both of these new observations resulted from our (improved) method of preparing the chromosomes for SEM and TEM.     

Azadirachta indica exhibits chemopreventive action against hepatic cancer: Studies on associated histopathological and ultrastructural changes

The present study was designed to evaluate the anticarcinogenic potential of Azadirachta indica against N-nitrosodiethylamine (NDEA)-induced hepatocarcinogenesis. Further, the associated histopathological and ultrastructural changes were also analyzed. Hepatic cancer model was developed by the intraperitoneal administration of NDEA to mice at weekly intervals, in successive increasing doses, for a period of 8 weeks. Aqueous A. indica leaf extract (AAILE) was administered orally at a dosage of 100 μg/g body weight thrice a week till termination of the study. A relationship between histopathological grading and chemopreventive effect of A. indica had been established at various stages of carcinogenesis. Anticancer activity of A. indica was evaluated in terms of tumor incidence, tumor multiplicity, and survival rate. A significant reduction in tumor incidence (33%), tumor multiplicity (42%), and increase in survival (34%) was observed upon administration of AAILE to NDEA-abused mice. Transmission and scanning electron microscopic investigations showed severe alterations in organelle organization, cellular arrangement, degree of differentiation, cellular metabolism, and morphology of the hepatocytes. These changes appeared to be distinctly delayed upon AAILE supplementation. The results suggest A. indica may have anticancer potential against NDEA-induced hepatic cancer.     

A new approach for improving the birefringence analysis of dental enamel mineral content using polarizing microscopy

The main problem in interpreting birefringence of dental enamel under polarizing microscopy is the lack of physical constants able to allow the Wiener equation to be applied directly to the composition of such tissue. The present study introduces a new approach to circumvent this constraint. Because the nonmineral phase of enamel is heterogeneous, its refractive index can be computed in terms of its components (namely, water, which is partially replaced by the immersion medium, and organic matter), thereby providing a more acceptable refractive index to be used in the Wiener equation. Furthermore, the enamel mineral volume is ordinarily calculated on the basis of the density 3.15 g cm^?3. The density 2.99 g cm^?3 has been, however, reported to be more accurate for enamel hydroxyapatite, so enamel mineral volumes from selected published data were converted using such a density. The birefringence of mature enamel computed by the Wiener equation, taking into account the above refinements, matched, for the first time, published experimental birefringence values. The theoretical water and organic contents were also consistent with published experimental data. Thus, a direct application of the Wiener equation to the enamel composition has now been achieved. It is speculated that quantitative data on the mineral, the water and the organic contents of mature dental enamel can be derived from interpretation of birefringence in two immersion media (obtained before and after extraction of the organic matter) with this new approach.