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PubMed2002

Scanning electron microscopy and energy-dispersive X-ray microanalysis studies of afibrillar cementum and cementicle-like structures in human teeth.

We examined afibrillar cementum (AFC) and cementicle-like structures (CLS) in human teeth by scanning electron microscopy and energy-dispersive X-ray microanalysis. The AFC showed a spur- or island-, plate- and mass-like structure with appositional laminations, while large masses in the enamel fissures enclosed CLS showing concentric appositional rings. Such AFC was observed in enamel fissures, an abnormal enamel pit, dens invaginatus and root furcations with enamel droplets, as well as on the cervical enamel surfaces, where ameloblasts are differentiated at the later or last stage of enamel formation. Cementicle-like structures were occasionally found independent from AFC and some CLS contained epithelial cell-like or ameloblast-like remnants in the core, surrounded by a few or many concentric rings. In addition, cementicles (CEC) in the root furcations also contained the remnants of Malassez's epithelial-rest cells surrounded by a few concentric rings. In some areas, AFC was mixed with enamel structures. These results show that the organic material in some parts of AFC and CLS may be derived from epithelial cells similar to that of CEC. Calcification values of AFC and CLS were significantly higher than that of fibrillar cementum, and the minute crystals are probably apatite.

Calcium Phosphates↗
PubMed1975

[New possibilities for demonstration of some biogenous amines in the adrenal by X-ray microanalysis (author's transl)].

Technical possibilities for the histochemical demonstration of some biogenous amines with the scanning electron-microscope and X-ray microanalysis were shown by using argentaffine and chromaffine reactions. Objects were adrenals of adult minipigs. Beside morphologic studies the amounts of silver or chromium can be determined semiquantitatively by the intensity of the specific X-radiate from "sum up-analysis" out of punctual areas. Furthermore the high amine concentrations in the adrenal medulla permit the performance of "line-analysis" and of "element-distribution-pictures" for silver or chromium. At the points chosen for analysis the concentration of epinephrine + norepinephrine is six times as high in the adrenal medulla as in the cortex; for norepinephrine alone it is only three times higher. Line-analysis shows that the concentration of katecholamines in the cortex is very low in the periphery and high near the medulla. The chromaffine reaction for serotonin leads to a distinct chromium peak in the medulla with a distribution quotient of 4.6 : 1 versus the cortex showing that there must be remarkable amounts of serotonin in the adrenal medulla, inspite of the fact that serotonin cannot by synthetized there from its natural precursor.

Adrenal Cortex↗
PubMed1999

Quantitative energy-dispersive X-ray microanalysis of calcium dynamics in cell suspensions during stimulation on a subsecond time scale: preparative and analytical aspects as exemplified with paramecium cells.

We analyzed preparative and analytical aspects of the dynamic localization of Ca(2+) during cell stimulation, using a combination of quenched flow and energy-dispersive X-ray microanalysis (EDX). Calcium (or Sr, as a substitute) was retained as fluorides during freeze-substitution, followed by epoxide embedding. The quenched-flow used allowed analyses, during stimulation, in the subsecond time range. Sections of 500 nm were analyzed and no artificial Ca or Sr leakage was recognizable. We calculated a primary beam spread from 63 to 72 nm that roughly indicated the resolution of EDX/structure correlation. These values are quite compatible with the size of potential structures of interest, e.g., Ca stores (approximately 100-nm thickness) or cilia (approximately 250-nm diameter). We used widely different standards to calibrate the ratio of CaK(alpha) net counts in relation to actual ¿Ca. Calibration curves showed a linear relationship and a detection limit of ¿Ca = 2 mM, while ¿Ca in cytosol was 3 mM and in stores was 43 mM, both in nonactivated cells. Eventually Sr(2+) can rapidly be substituted for Ca(2+) in the medium before and during stimulation, thus allowing one to determine Me(2+) fluxes. With our "model" cell, Paramecium, we showed that, upon stimulation (causing rapid Ca(2+) mobilization from subplasmalemmal stores), Ca was immediately exchanged for Sr in stores.

Animals↗
PubMed1996

Chloride secretion in the submandibular gland of adult and early postnatal rats studied by X-ray microanalysis.

Submandibular acinar cells of 1-day-old, 7-day-old, and adult rats were analyzed with X-ray microanalysis after stimulation with carbachol for different time periods (2-7 min). In unstimulated animals, marked differences in elemental content between compartments could be observed: secretory granules had a higher Ca and lower P and K content than other cell compartments. Comparison between different age groups showed significant differences for Ca, which increased with age in all compartments; Mg increased with age in the secretory granules and the apical cytoplasm. Only the glands from adult animals showed a significant effect of cholinergic stimulation: a transient decrease in Cl and K. The Cl concentration in the secretory granules decreased to 60% of the control value, which suggests that the granules release Cl upon stimulation. In young animals, no or little change in elemental distribution was observed after stimulation. This may indicate that Cl- secretion mechanisms are much less prominent in young animals. The ultrastructure of submandibular secretory granules depends on the preparation method: condensed and electrondense in freeze-substituted unfixed tissue, decondensed and more translucent in aldehyde-fixed tissue. This may indicate that the granules can transport water, and swell during the process of aldehyde fixation.

Aging↗
PubMed1981

Biomineralization on crinoid echinoderms. Characterization of crinoid skeletal elements using TEM and STEM microanalysis.

Columnals of Neocrinus blakei, a modern species of stalked crinoid, were studied using a variety of analytical techniques. Analyses of the magnesium calcite of the crinoid stereom using powder X-ray diffraction and electron microprobe analysis yield a composition of Ca 88Mg 12C03. Scanning transmission electron microscopy (STEM) microanalytical data indicate that Mg incorporation into the calcite structure of the crinoid stereom is random and homogeneous to at least the 20 nm level. There appear to be no variations in composition at this level either within or between structural entities of the crinoid columnal stereom. TEM reveals a heterogeneity of contrast which may be due to incorporation of organic material or some other substance which is non-crystalline in character. Single-crystal X-ray diffraction data indicate that the individual skeletal plates are single crystals which yield diffuse and imperfect X-ray reflections due to a mosaic structure. Subsequent selected area electron diffraction (SAD) photographs via TEM, using various sizes of SAD apertures, indicate that the crystallites making up the mosaic structure are (order of magnitude) about 1.0 micrometer in size. The presence of mosaic structure in the single crystal skeletal elements may at least in part explain the lack of cleavage in fracture surfaces of echinoderm skeletal material. Based on these data, as well as data from skeletal elements of other deep water, stalked crinoids, we feel that these results may be applicable to crinoids in general, at least those existing in relatively constant temperature environments. The single-crystal nature of crinoid high magnesium calcite, and its remarkable homogeneity of composition suggest that a large "vital effect" (i. e., biologic control of skeletal deposition) mediates the mineralization process.

Animals↗
PubMed1992

High resolution electron microscopic investigation of atomic structural images in human atherosclerotic plaque.

Atomic structural images in the calcified matrix of human atherosclerotic plaques were characterized by high-resolution electron microscopy (HREM). The mineral elements distributed in the calcified matrix were analysed by energy dispersive X-ray microanalysis. Careful HREM observations revealed that a large number of microcrystallites and crystal particles with a broad spectrum of crystal size and perfection were embedded in the amorphous matrix of the calcified plaques. The amorphous phase in the calcified matrix was made up of a dense network of granulo-fibrillar substructures. The lattice-fringes with regular arrangement of atom columns along the lattice arrays were clearly demonstrated in the crystalline particles of the calcified plaques. A number of characteristic granulo-fibrillar substructures in the amorphous phase could be seen to be tightly connected to the lattice-fringes in the crystalline phase along the interfaces between the amorphous and crystalline phases. The crystal lattices at the boundary between two crystals were very well matched with the periodicity of visible atoms continuous through the two crystals. The mineral elements such as calcium and phosphorus were detected to be diffusely distributed in the calcified matrix of the atherosclerotic plaques. Based on these findings, it is concluded that: (1) an interlacing network of granulo-fibrillar substructures in the amorphous matrix may play a structural and/or regulatory role for the organic matrix by providing sites for nucleation; and (2) the deposition of the mineral elements may be effective for the transformation of the amorphous nucleation phase to well-crystallized nucleation phase.

Aorta↗
PubMed1988

Rat lung type II cell and lamellar body: elemental composition in situ.

We determined the in situ elemental composition of alveolar type II cells (ATII) and lamellar bodies (LB) with electronprobe microanalysis (EPMA) of freeze-dried unstained cryosections (100-200 nm) obtained from lungs frozen in anesthetized rats. Twenty-nine ATII from seven rats were subjected to EPMA. Cytoplasmic (Cyto) composition was the following (in mmol/kg dry wt, mean +/- SE, n = 30): 136 +/- 14.1 Na, 60 +/- 2.8 Mg, 549 +/- 34.8 P, 278 +/- 10.5 S, 158 +/- 7.3 Cl, 525 +/- 26.4 K, and 6.6 +/- 0.9 Ca. LB composition was the following (n = 66): 44 +/- 4.0 Na, 7.9 +/- 0.8 Mg, 1,060 +/- 25.0 P, 79 +/- 4.8 S, 64 +/- 3.6 Cl, 114 +/- 4.1 K, and 30 +/- 0.9 Ca. P and S concentrations were consistent with previous biochemical determinations of phospholipid and protein content of isolated LBs. LBs contain significantly more Ca and less Mg than Cyto. Ca correlated significantly with LB P but not S concentration, and the reported low Ca binding affinity of similar phospholipid mixtures implies a high LB free Ca concentration. Ca was significantly higher in apical and exocytotic LBs compared with those in the perinuclear region. Differences between LB and Cyto monovalent ion concentrations are not entirely due to the difference in hydration revealed by significantly lower K-Cl ratios in LBs. The relative excess of Cl and Ca in LB suggests that these ions may be distributed by active transport systems known to be present in the Golgi apparatus and in Golgi-derived organelles of other cell types.

Analysis of Variance↗
PubMed1996

Bone bonding ability of an apatite-coated polymer produced using a biomimetic method: a mechanical and histological study in vivo.

A 20-microns thick apatite layer was coated onto polyethersulfone (PES) rectangular plates by soaking them in simulated body fluid containing CaO-SiO2 based glass powder. Coated and uncoated PES plates (10 x 15 x 1.5 mm) were implanted in the tibiae of rabbits, which were sacrificed 8, 16, and 30 weeks thereafter, and the samples were examined histologically using contact microradiography (CMR), Giemsa surface staining, and a scanning electron microscope connected to an electron probe microanalyzer (SEM-EPMA). The tensile failure loads at the bone/implant interfaces were determined using the detaching test. The histological examinations showed excellent bone apposition on coated PES and the sign of degradation of the apatite layer at remodeling lacunae. The apatite layer underwent complete resorption and was replaced by bone in most areas of the bone/implant interface after 30 weeks. Bone did not bond directly to uncoated PES after each follow-up period. The failure loads between bone and coated PES at 8, 16, and 30 weeks after implantation were 1.7 +/- 0.35, 2.36 +/- 0.53, and 1.45 +/- 0.48 kg, respectively. Those between bone and uncoated PES were nearly 0 kg at each postimplantation period. Failure during the detaching test occurred at the bone/apatite interface or near it after 8 weeks. After 16 weeks, it usually occurred at the apatite/ PES interface or near it, and occasionally in the middle of the apatite layer. The apatite layer was hardly detected at the failured interface after 30 weeks. In this study, an apatite-coated PES produced using a biomimetic method was demonstrated to bond directly to bone without any intervening soft tissue, which indicates that this material possesses excellent bioactivity.

Animals↗
PubMed1996

Bone bonding behavior of titanium and its alloys when coated with titanium oxide (TiO2) and titanium silicate (Ti5Si3).

It has been proposed that the essential requirement for artificial materials to bond to living bone is the formation of bonelike apatite on their surfaces in the body. Recent studies have shown that titanium hydrogel and silica gel induce apatite formation on their surface in a simulated body fluid. In this study, the influence of titanium oxide and titanium silicate on the bonding of titanium alloys to bone was studied. Rectangular implants (15 x 10 x 2.2 mm) of titanium, Ti-6Al-4V, Ti-6Al-2Nb-Ta, Ti-6Al-4V coated with TiO2, and Ti-6Al-4V coated with Ti5Si3 were implanted into the tibial metaphyses of mature rabbits. At 8 and 24 weeks after implantation, the tibiae containing the implants were dissected out and subjected to a detaching testing. The failure load for titanium, Ti-6Al-4V, Ti-6Al-2Nb-Ta, Ti-6Al-4V coated with TiO2, and Ti-6Al-4V coated with Ti5Si3 were, respectively, 0.68 +/- 0.48, 0.22 +/- 0.46, 0.67 +/- 0.59, 2.18 +/- 0.71 and 2.03 +/- 0.41 kgf at 8 weeks, and 2.7 +/- 0.91, 2.58 +/- 1.29, 2.38 +/- 0.41, 3.79 +/- 1.7, and 2.79 +/- 0.87 kgf at 24 weeks after implantation. Histological examination by Giemsa surface staining, CMR, and SEM-EPMA revealed the coated titanium alloy implants directly bonded to bone tissue during early implantation. A Ca-P layer was observed at the interface of the coated implants and the bone. The results of this study indicated that TiO2 and Ti5Si3 can enhance the early bonding of titanium alloys to bone by inducing a Ca-P layer (chemical apatite) on the surface of titanium alloys. It also is suggested that the direct bone contact occurs in relation to the calcium and phosphorus adsorption onto the surface of the titanium passive layer formed during long-term implantation.

Alloys↗
PubMed1998

Analysis of titanium dental implants after failure of osseointegration: combined histological, electron microscopy, and X-ray photoelectron spectroscopy approach.

A multitechnique approach has been used to characterize the surface of nonosseointegrated titanium implants and the surrounding biological tissues. Five pure titanium dental implants were used as reference, and 25 removed implants were studied. Surface and in-depth chemical compositions of the implants (from a total of 16 patients) were investigated by X-ray photoelectron spectroscopy (XPS). Histological slides of the surrounding tissues were examined by light microscopy, XPS, and electron microprobe analysis. None of the failed implants presented the regular surface composition and depth profile of the TiO2 overlayer; foreign elements (Ca, Na, P, Si, Cl, Zn, Pb, and Al) were observed on some implants. Fibrosis, lymphocytic and plasmocytic infiltrates, and granulomatous lesions were detected in the surrounding tissues. XPS and electron microprobe analysis indicated the presence of Zn, Fe, Sn, and Ti in the tissues. As a possible scenario for implant failure, we propose and discuss a oxidoreduction mechanism, leading to a partial dissolution or the complete dissociation of the protective titanium dioxide overlayer and to ion diffusion through the surrounding tissues.

Connective Tissue↗
PubMed1999

Optimal conditions for alumina coating formation on the MA956 superalloy for prosthetic bearing applications.

An experimental study of the oxidation treatment at high temperature of the ODS MA956 superalloy was conducted in an attempt to achieve a protective alumina scale for biomedical applications. A quadratic response-surface model was developed in order to study the effects of treatment time and temperature (in the range of 1000 degrees C to 1250 degrees C) on scale thickness. The obtained model adequately represents the experimental response and shows that the thickness gradients of the layer increase with the temperature for each exposure time and decrease steadily to zero as the treatment time increases. The microstructural characterization reveals that the alumina scale formed at or above 1000 degrees C consists of an alpha-alumina phase. Treatments at temperatures above 1150 degrees C give rise to an alumina scale with some defect probability. An increase in the temperature up to 1200 degrees C gives rise to the appearance of some blistering of the superficial scale. An oxidation treatment of 100 h at 1100 degrees C was found to be the best for guaranteeing the formation of a defect-free, compact, adherent, and continuous alpha-alumina scale thick enough to support satisfactory wear and biological conditions.

Alloys↗
PubMed1987

Ultrastructure of cardiocyte degeneration and myocardial calcification in the dystrophic hamster.

The myocardium of the Bio 14.6 cardiomyopathic hamster was examined with the electron microscope to identify cellular and organelle changes during the acute lesioning stage, a period typified by concomitant cardiocyte destruction and calcium elevation. Most cardiocytes retained their normal histologic and ultrastructural features, but scattered foci of altered and necrotic cells were observed in association with degenerative calcifying lesions. Prenecrotic alterations of myocytes included cellular edema; varying degrees of distension of sarcoplasmic reticulum and T-tubules; contraction bands and other myofibrillar abnormalities; mitochondrial clustering and hyperplasia; a wide spectrum of mitochondrial changes such as altered sizes, shapes, and cristal patterns, and increases in the number and size of osmiophilic matrix inclusions. Morphologic features consistent with substantial calcium excess were not observed in most altered but prenecrotic cells. Instead, calcium deposition within extruded mitochondria and upon degenerating organelle debris was observed only after cardiocyte disruption. Some calcifying cell remnants were phagocytized by macrophages, whereas large calcified plaques and other deposits remained in the interstitium. Mitochondrial calcification in vascular smooth muscle cells and fibroblasts was evident in highly calcified lesions. These observations suggest that most of the morphologically identifiable calcium deposition present in this cardiomyopathy results from secondary calcification subsequent to sarcolemmal disruption.

Age Factors↗
PubMed1990

Organization and development of the mineral phase during early ontogenesis of the bony fin rays of the trout Oncorhynchus mykiss.

Characterization of mineral deposition has been studied by electron optical methods during early ontogenesis of lepidotrichia, the bony fin rays, of the trout Oncorhynchus mykiss (the former Salmo gairdneri). The fin rays consist of an extracellular granular ground substance containing in part a network of collagen fibrils within the basal lamella of the fin dermoepidermal interface. Growth of individual rays proceeds in a proximodistal direction. The mineral phase appears as electron-dense needle or plate-like particles and is associated with the collagenous matrix. On analysis of progressively maturing tissue, the mineral was characterized as a poorly crystalline hydroxyapatite with Ca/P molar ratios in the range of 1.0-1.4, corresponding to distal and proximal areas, respectively. With selected-area electron diffraction and dark field imaging of lepidotrichia, the mineral particles were found to be about 3-10 nm thick and 12-20 nm in length (along their crystallographic c-axes), possibly aggregated into larger crystals 35-40 nm long observed with bright field microscopy. No definitive relation was found between either the c- or a,b-axes images of the crystals and the periodic structure of collagen, which forms the framework for mineral deposition in this and in other vertebrate calcifying tissues.

Animals↗
PubMed1991

Effect of hydrophilic soft segment side chains on the surface properties and blood compatibility of segmented poly(urethaneureas).

Segmented poly(urethaneureas) with hydrophilic side chains were prepared from poly(tetramethylene oxide) (PTMO), 4,4'-diphenylmethane diisocyanate (MDI), ethylene diamine (ED) and a diol with a long hydrophilic side chain comprised of an ethylene oxide-proplene oxide copolymer. The end groups of the hydrophilic chains were either sodium sulfonate or methoxy groups. The state of microphase separation showed a small dependence on the fraction of long-chain hydrophilic diol. Surface analysis by means of static underwater contact angle and dynamic contact angle measurements revealed that the graft chains were at the aqueous interface in the hydrated state. An ex vivo A-V shunt experiment revealed that a more thrombogenic blood-material response was correlated with an increase in the concentration of polymeric hydrophilic side chain incorporation. The polyurethane containing a long chain diol with methoxy end groups exhibited a higher level of thrombogenicity than the similar polymers possessing a sulfonate terminated side chain.

Animals↗
PubMed2003

SEM/EDS comparison of polar and seasonal temperate ice.

We have developed a method employing a scanning electron microscope (SEM) and energy dispersive X-ray spectroscopy for examining uncoated ice specimens. By permitting the ice to sublimate in the SEM at temperatures between -115 degrees and -60 degrees C, enough ions are produced to prevent specimen charging. The absence of a conductive coating permits both the acquisition of uncompromised X-ray spectra, and a dynamic view of impurities as the ice sublimates. The method has enabled us to examine the microstructure and impurities in ice in ways not possible through standard melt chemistry measurements or even through using a SEM to study coated samples. Soluble impurities appeared either as white spots in grain interiors, grain boundaries, and triple junctions, or as filaments in grain boundaries. Inclusions of insoluble impurities have also been observed in natural ice. Thus, we have been able to compare the microstructural location and concentration of impurities in ice from different terrestrial locations. Even when ion chromatography of the melt from two core sections reveals similar levels of impurities, the morphology and location of the impurity aggregates can be quite different. Analysis of impurity type and location can provide clues to the depositional environment and history of the ice.

Cold Climate↗
PubMed1987

Some nanostructural features in ceramics.

Nanostructural features in some ceramics have been discussed and reviewed. Based on our research results and recent published investigations, many topics, such as grain, grain boundary, interface film, grain boundary engineering, microcrack, microdomain, nanodomain, domain boundary, and phase transformation, etc., have been dealt with; and many materials, such as Si3N4, beta''-Al2O3, MgO, SiC, (Hg, Cd) Te, BNN, ZrO2, PLZT, CdSe, Ca10(PO4)6, (OH)2, etc., have been involved. The results are important to understand the relation between the structure and property of materials and to improve the materials' technology.

Ceramics↗