Electron microscopy analysis of the thermal phase transition from hydroxyapatita to ß-TCP observed in human teeth.

Samples of enamel and dentin from human molar teeth were heated in air from room temperature (25°C) up to 1200°C and the phase transition from hydroxyapatite (HAP) to tricalcium phosphate (ß-TCP) was recorded. The changes produced in morphology and chemical composition in the tooth during heating were analysed by light microscopy, scanning electron microscopy (SEM), characteristic x-ray energy dispersion spectroscopy (EDS), x-ray diffraction (XRD), electron diffraction, transmission electron microscopy (TEM) and high-resolution electron microscopy (HRTEM). The results indicated a high correlation relationship among Ca content, P content, O content and Na content, and the existence of the Kirkendall effect during the HAP- ß-TCP phase transition. LAY DESCRIPTION: This work is related with tooth materials. Samples of enamel and dentine from human molar teeth were heated in air from 25°C up to 1200°C and the phase transition from hydroxyapatite (HAP) to tricalcium phosphate (ß-TCP) was recorded. The ß-TCP phase is also known as whitlockite. The changes produced in morphology and chemical composition in the tooth during heating were analysed by electron microscopy and X-ray diffraction. The results indicated a high correlation relationship among Ca, P, O and Na contents, and the existence of the Kirkendall effect, the atomic diffusion producing voids, during the HAP- ß-TCP phase transition.

Helical Growth of Ultrathin Gold-Copper Nanowires.

In this work, we report the synthesis and detailed structural characterization of novel helical gold-copper nanowires. The nanowires possess the Boerdijk-Coxeter-Bernal structure, based on the pile up of octahedral, icosahedral, and/or decahedral seeds. They are self-assembled into a coiled manner as individual wires or into a parallel-ordering way as groups of wires. The helical nanowires are ultrathin with a diameter of less than 10 nm and variable length of several micrometers, presenting a high density of twin boundaries and stacking faults. To the best of our knowledge, such gold-copper nanowires have never been reported previously.

Aberration-Corrected Transmission Electron Microscopic Study of the Central Dark Line Defect in Human Tooth Enamel Crystals.

Angstrom resolution images of human tooth enamel (HTE) crystallites were obtained using aberration-corrected high-resolution transmission electron microscopy and atomic-resolution scanning transmission electron microscopy in the modes of bright field, annular dark field, and high-angle annular dark-field. Images show that the central dark line (CDL) defect observed around the center of the HTE crystals is a site for caries formation in the HTE and has a thickness of ~0.2 nm. Results also suggest that the CDL goes through one of the OH- planes.

Evidence of noncentrosymmetry of human tooth hydroxyapatite crystals.

Herein, we investigate human single hydroxyapatite crystals (enamel and dentine) by convergent-beam electron diffraction (CBED) and automated electron-diffraction tomography (ADT). The CBED pattern shows the absence of the mirror plane perpendicular to the c axis leading to the P63 space group instead of the P63 /m space group considered for larger-scale crystals, this is confirmed by ADT. This experimental evidence is of prime importance for understanding the morphogenesis and the architectural organization of calcified tissues.

Presence of matrix vesicles in the body of odontoblasts and in the inner third of dentinal tissue: a scanning electron microscopic study.

OBJECTIVES: The aim of this report is to present the results of a scanning electron microscopic study on the presence of matrix vesicles (MVs) found in human dentine. STUDY DESIGN: Dentin tissue from 20 human bicuspids was analyzed by means of scanning electron microscopy. RESULTS: MVs were found as outgrowths of the cellular membrane of the odontoblastic body, the more proximal portion of the odontoblastic process before entering the dentinal tubule and in the odontoblastic process within the inner third of the dentin. Size of MVs varied depending on location. In the inner third of dentin, they were seen in diverse positions; as membranal outgrowths, deriving from the odontoblastic process, lying free in the intratubular space and attached to the dentinal wall. Sometimes, they were seen organized forming groups of different sizes and shapes or as multivesicular chains running from the surface of the odontoblastic process to the tubular wall. MVs were present in places never considered: 1) the body of odontoblasts; 2) the most proximal part of the odontoblastic processes before entering the circumpulpal dentine and also: 3) in the inner third of dentinal tissue. CONCLUSIONS: According to our results, MVs not only participate during mantle dentin mineralization during early dentinogenesis, they also contribute during the mineralization process of the inner dentin.

An approach to investigate the crystallographic unit cell of human tooth enamel.

Human tooth enamel (HTE) is the hardest tissue in the human body and its structural organization shows a hierarchical composite material. At the nanometric level, HTE is composed of approximately 97% hydroxyapatite [HAP, Ca10(PO4)6(OH)2] as inorganic phase, and of 3% as organic phase and water. However, it is still controversial whether the hexagonal HAP phase crystallizes in P63/m or another space group. The observance in HTE of Ca2+, Mg2+ and Na+ ions using X-ray characteristic energy-dispersive spectroscopy in the scanning electron microscope has been explained by substitutions in the HAP unit cell. Thus, Ca2+ can be replaced by Na+ and Mg2+ ions; the PO43- group can be replaced by CO32- ions; and the OH- ions can also be replaced by CO32-. A unit-cell model of the hexagonal structure of HTE is not fully defined yet. In this work, density functional theory calculations are performed to study the hexagonal HAP unit cell when substitution by OH-, CO32-, Mg2+ and Na+ ions are carried out. An approach is presented to study the crystallographic unit cell of HTE by examining the changes resulting from the inclusion of these different ions in the unit cell of HAP. Enthalpies of formation and crystallographic characteristics of the electron diffraction patterns are analysed in each case. The results show an enhancement in structural stability of HAP with OH defects, atomic substitution of Mg2+, carbonate and interstitial Na+. Simulated electron diffraction patterns of the generated structures show similar characteristics to those of human tooth enamel. Hence, the results explain the indiscernible structural changes shown in experimental X-ray diffractograms and electron diffraction patterns.

Evaluation of crystalline indexes obtained through infrared spectroscopy and x-ray diffraction in thermally treated human tooth samples.

The crystalline indexes obtained through infrared spectroscopy (CIIR) and X-ray diffraction (CIXRD) were used to analyze the structural changes of dentin and enamel in human teeth when subjected to heat treatments between room temperature (25 °C) to 1200 °C in atmospheric air and argon. Thermogravimetric (TGA) and differential scanning calorimetry (DSC) analyses of sound powders were also used to support the analysis. As the temperature increased, the CIXRD increased exhibiting a behavior like that of the crystal grain size, and the CIIR increased until the ß-TCP phase appeared. These results indicated that the CIIR was more variable to structural changes than the CIXRD.

The SEM electron-mirror effect in human tooth and synthetic hydroxyapatite samples.

The characteristics of the electron-mirror effect (EME) image depend on both the scanning electron microscope parameters and the sample's physical properties. The behavior of human tooth (dentin and enamel) and synthetic hydroxyapatite samples submitted to the EME procedure is presented in this work. Polyethylene terephthalate (PET) and epoxy resin, two good EME producers, were used for comparison. A distorted EME image was observed in the obtained dentin's surface, but enamel and synthetic hydroxyapatite surfaces did not produce the EME. After ex situ calcination treatments of the teeth at 700 and 1,200°C, the EME was observed in dentin, enamel, and synthetic hydroxyapatite, but highly deformed EME images were produced. We show that these last observations are the result of the well-known charge-edge effect. After EME analysis, the calculated dielectric constant was 8.7 for dentin and 3.8 for PET. RESEARCH HIGHLIGHTS: Electron-mirror effect (EME) was observed in dentin but not in enamel or synthetic hydroxyapatite. Highly deformed EME images are produced in all samples after calcination at above 700°C. For dentin the calculated dielectric constant was 8.7 and for PET is was 3.8.

Elaboration and structural analysis of aquasomes loaded with indomethacin.

The aim of this study was to prepare nanoparticles in form of aquasomes with Indomethacin as a low solubility drug mode. Aquasomes charged with Indomethacin were obtained through the formation of an inorganic core of calcium phosphate covered with a Lactose film and further adsorption of the Indomethacin. Structural analyses, particle size, and morphology were evaluated by X-ray powder diffractometry, transmission electron microscopy, and scanning electron microscopy. The X-ray analysis of the samples and their observation through electronic microscopy allowed us to identify the inorganic calcium phosphate nucleus formation, as well as the layers of Lactose and Indomethacin. The particle size analysis of the aquasomes obtained with the Lactose layer and charged with the drug indicated an average particle size in the range of 60-120 nm, with a media of 90 nm. Standard deviation was 18.0234 and the standard error of the media 4.1348. The method was reproducible under the conditions used to prepare the aquasomes, such as ultrasound frequency and the moment of sonication for the formation of inorganic cores.

Scanning electron micrographic features of a giant submandibular sialolith.

To recognize recently appearing mineralization phenomena, one must study the external surface of the sialoliths, since it is not possible to study them in the central portions of sialoliths. The authors examined the external surface of a sialolith by scanning electron microscopy and analyzed its microstructures. The study revealed the presence of numerous microstructures of different shapes (nodular, laminar, reticular, microgranular, and multinodular) and variable size arranged in a haphazard fashion. The diverse microstructures encountered strongly suggest that different mechanisms of mineralization occur during growth and development of the sialoliths.

Crystal structure characterization of nautilus shell at different length scales.

In this work, we studied the shell structure of the Nautilus pompilius Linnaeus by using analytical techniques of scanning (SEM) and transmission electron microscopy (TEM) and X-ray diffraction. The main objective of this study is the structural characterization of Nautilus shell at different length levels, from micron to nano-scale. The results were also used to try to determine the shell structure mechanism of formation. The information obtained in this work will place our particular knowledge a closer step to understand how self-assembly works in nature, and will increase the opportunities of using this information in the future synthesis of new advanced materials.

Scanning Electron Microscopic Study on the Fibrillar Structures within Dentinal Tubules of Human Dentin.

INTRODUCTION: Pulp biology is central to the whole tooth, and knowledge on its microstructure is changing with new studies. This study presents certain microfibrillar structures found within the dentin tubules of human teeth connecting dentin tubules and odontoblastic processes. METHODS: We analyzed the crowns of 30 noncarious, human teeth. They were fixed; demineralized; and, later, processed and reviewed by means of scanning electron microscopy. RESULTS: In the predentin layer, we found numerous fine fibrillar structures connecting the odontoblastic process and the wall of the dentinal tubule. In the inner dentinal third, we observed structures forming a dense microfibrillar network of variable thickness and diameters. These microstructures were very thin and numerous in this area, and their number decreased as more external dentin levels were examined. CONCLUSIONS: According to the review of the literature and our findings, these microfibrillar structures may be an unrecognized support system that holds and secures the odontoblastic process within the dentinal tubule.

Wettability modification of human tooth surface by water and UV and electron-beam radiation.

The wettability of the human tooth enamel and dentin was analyzed by measuring the contact angles of a drop of distilled water deposited on the surface. The samples were cut along the transverse and longitudinal directions, and their surfaces were subjected to metallographic mirror-finish polishing. Some samples were also acid etched until their microstructure became exposed. Wettability measurements of the samples were done in dry and wet conditions and after ultraviolet (UV) and electron beam (EB) irradiations. The results indicate that water by itself was able to increase the hydrophobicity of these materials. The UV irradiation momentarily reduced the contact angle values, but they recovered after a short time. EB irradiation raised the contact angle and maintained it for a long time. Both enamel and dentin surfaces showed a wide range of contact angles, from approximately 10° (hydrophilic) to 90° (hydrophobic), although the contact angle showed more variability on enamel than on dentin surfaces. Whether the sample's surface had been polished or etched did not influence the contact angle value in wet conditions.

Gold-copper nanostars as photo-thermal agents: synthesis and advanced electron microscopy characterization.

Nanoalloys have emerged as multi-functional nanoparticles with applications in biomedicine and catalysis. This work reports the efficient production and the advanced transmission electron microscopy characterization of gold-copper pentagonal nanostars. The morphology of the branches is controlled by the adequate choice of the capping agent. When oleylamine is used rounded nanostars are produced, while pointed nanostars are obtained by using hexadecylamine. Both types of nanostars were proved to be thermally stable and could therefore be used as therapeutic agents in photo-thermal therapies as confirmed by the near-infrared absorption spectra.

XRD and FTIR crystallinity indices in sound human tooth enamel and synthetic hydroxyapatite.

The crystallinity index (CI) is a measure of the percentage of crystalline material in a given sample and it is also correlated to the degree of order within the crystals. In the literature two ways are reported to measure the CI: X-ray diffraction and infrared spectroscopy. Although the CI determined by these techniques has been adopted in the field of archeology as a structural order measure in the bone with the idea that it can help e.g. in the sequencing of the bones in chronological and/or stratigraphic order, some debate remains about the reliability of the CI values. To investigate similarities and differences between the two techniques, the CI of sound human tooth enamel and synthetic hydroxyapatite (HAP) was measured in this work by X-ray diffraction (XRD) and Fourier Transform Infrared spectroscopy (FTIR), at room temperature and after heat treatment. Although the (CI)XRD index is related to the crystal structure of the samples and the (CI)FTIR index is related to the vibration modes of the molecular bonds, both indices showed similar qualitative behavior for heat-treated samples. At room temperature, the (CI)XRD value indicated that enamel is more crystalline than synthetic HAP, while (CI)FTIR indicated the opposite. Scanning (SEM) and transmission (TEM) images were also used to corroborate the measured CI values.

Human Cementum Protein 1 induces expression of bone and cementum proteins by human gingival fibroblasts.

We recently presented evidence showing that a human cementoblastoma-derived protein, named Cementum Protein 1 (CEMP1) may play a role as a local regulator of cementoblast differentiation and cementum-matrix mineralization. This protein was shown to be expressed by cementoblasts and progenitor cells localized in the periodontal ligament. In this study we demonstrate that transfection of CEMP1 into human gingival fibroblasts (HGF) induces mineralization and expression of bone and cementum-matrix proteins. The transfected HGF cells had higher alkaline phosphatase activity and proliferation rate and they expressed genes for alkaline phosphatase, bone sialoprotein, osteocalcin, osteopontin, the transcription factor Runx2/Cbfa1, and cementum attachment protein (CAP). They also produced biological-type hydroxyapatite. These findings indicate that the CEMP1 might participate in differentiation and mineralization of nonosteogenic cells, and that it might have a potential function in cementum and bone formation.

Anti-cementoblastoma-derived protein antibody partially inhibits mineralization on a cementoblastic cell line.

The effect of human anti-cementoblastoma-derived protein antibody during cementogenesis in vitro was investigated by using human cementoblastoma-derived cells. Cultures treated with 5 microg/ml of CP antibody from day 1 to day 15 revealed a significant decrease in alkaline phosphatase activity (ALP) 40% (p < 0.005), 44% (p < 0.001), 49% (p < 0.1), and 45% (p < 0.02) at 9, 11, 13, and 15 days, respectively. Immunoexpression of osteopontin revealed that in cultures treated with anti-CP antibody, the positive number of cementoblastoma cells was reduced by 87, 83, 69, and 52% at 5, 7, 9, and 11 days, respectively. Bone sialoprotein immunoexpression showed a decrease in positive cells of 82, 51, 60, 80, 83, and 87% at 5, 7, 9, 11, 13, and 15 days, respectively, as compared to controls. The Ca/P ratio of the mineral-like tissue deposited in vitro by cementoblastoma cells revealed that control cultures had a Ca/P ratio of 1.45 and 1.61 at 5 and 15 days, whereas experimental cultures revealed a Ca/P ratio of 0.50 and 0.79 at 5 and 15 days, respectively. Electron diffraction patterns showed inner double rings representing D-spacing that were consistent with those of hydroxyapatite in both control and experimental cultures. Examination of the crystallinity with high resolution transmission electron microscopy showed homogeneous and preferential spatial arrangement of hydroxyapatite crystallites in control and experimental cultures at 15 days. Atomic force microscopy images of control cultures at 5 and 15 days revealed small granular particles and grain agglomeration that favored the formation of crystalline plaques with a lamellar-like pattern of the mineral-like tissue. Experimental cultures at 5 and 15 days showed tiny and homogeneous granular morphology. The agglomerates maintained spherical morphology without organization of needle-like crystals to form plaque-like structures. Based on these findings, it is hypothesized that cementoblastoma-derived protein may be associated to crystal growth, compositional and morphological features during the mineralization process of cementum in vitro.