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Electron probe microanalysis of isolated brain capillaries poisoned with lead.

The blood-brain barrier has been proposed as an important site for the toxic action of lead in the central nervous system. To investigate this, capillary endothelial cells were isolated from rat cortex and exposed to lead in vitro. Tissue suspensions were then prepared for electron microscopy and X-ray microprobe analysis. In cells exposed in vitro to lead, electron-dense deposits were observed within mitochondria. With X-ray analysis, it was determined that these intramitochondrial deposits contained lead in a non-crystalline matrix. Also, lead appeared to be accumulated in the same intramitochondrial areas as calcium. The results suggest that lead is preferentially sequestered in mitochondria of capillary endothelial cells. Further, this selective localization may be associated with lead-induced disruptions in intracellular calcium metabolism and transepithelial transport processes.

Animals↗

Mineralization of human aortas with coarctation: quantitative electron probe microanalysis.

Aortas with coarctation (isthmus stenosis) are obviously an ideal model to investigate pressure dependent changes of the aorta, as one can compare the proximal region (high pressure) with the distal region (low pressure). 7 aortas of patients aged from 2 months to 54 years were investigated. The concentrations of Na, Mg, P, S, C1, K, and Ca were determined by electronprobe microanalysis. Ca and P are constituent parts of the developing mineral and are mainly discussed. The Ca/P ratio (by weight) is about 0.3 in the unmineralized and about 2 in the mineralized tissue. Mineralization is demonstrated by a parallel increase of Ca, P, and the Ca/P ratio. Enrichments of Ca and P are found above all in a subintimal band. Usually they occur proximally and distally, but are much more pronounced proximally. Even in the aorta of a 2 month old infant enrichments were found proximally, but not distally. Thus mineral deposits occur very early in regions of hypertension. The differences between the proximal and distal region may demonstrate the influence of blood pressure on vascular mineralization. The mineralization starts in small compartments, which increase in size and number in the process of mineralization. Mineralized regions could often be localized by cathodoluminescence. But to demonstrate and localize the initial mineral deposits and to quantify element contents, electronprobe microanalysis became indispensable.

Adolescent↗

31P-NMR study of P-based dental adhesives and electron probe microanalysis of simulated interfaces with dentin.

The structure and function of P in P-based dental adhesives have not been defined properly. The purpose of this study was to evaluate these parameters in some currently available materials. Five dental adhesives were selected: Bondlite, Clearfil New Bond, J&J Dental Adhesive, Prisma Universal Bond, and Scotchbond. High-resolution 31P-NMR spectroscopy was used to identify the phases of the incorporated P and the kinetics of monomer-bonded P in an aqueous environment. Areas of 5 mm diameter located on polished sound dentin were subjected to the corresponding adhesive treatments and covered with translucent molds of the same diameter, which were filled up to 1 mm length with a visible-light-cured hybrid composite. Following 60 sec of light exposure from lateral directions, the specimens were embedded in fast-setting resin, cross-sectioned, and analyzed in an electron probe microanalyzer. The elemental profiles of Ca, P, and Cl were determined from area and line scans. Great differences in the 31P resonance shifts were obtained from the tested adhesives, indicating the presence of monomer-bonded and free P. A rapid hydrolysis of the resin-bonded P in the presence of water was identified. The x-ray images revealed dentin demineralization, with distinct differences in the P distribution pattern and the P/Cl ratio.

Dental Bonding↗

Electron probe microanalysis of fluorotic bovine teeth.

Incisor teeth were obtained from adult cattle which since 4 months of age to 5 or 6 years were maintained on rations containing a yearly average of 40 ppm F in the forage. Microchemical analyses were performed on the fluorotic bovine incisors. The microdistribution of fluoride varied markedly at different sites within the same tooth. Fluoride concentrations varied with the depth from the tooth surface and were influenced by the concentrations of fluoride present in the forage during amelogenesis, and the presence of hypoplastic pits and hyperplastic coronal cementum in enamel. The cementum in these lesions contained remarkably high concentrations of fluoride, and it was less calcified and more porous than adjacent enamel.

Animals↗

[Electron probe microanalysis of fluorine content in deciduous and permanent teeth from an area with a fluoride-deficient (0,3% mg/I) water supply].

The content of fluorine in deciduous and permanent teeth of young people was determined by x-ray microanalysis. The purpose of this investigation was to determine the concentrations of F in teeth without any supplementation of fluorides and to elaborate basic dates as a standard of comparison. F was measured in microvolumes with a wavelength-dispersive spectrometer (Step-Scan 100 sec) after coating with about 300 A of Carbon. The highest values were established in the outermost surface of enamel with quantities between 300 and 600 micrograms/g. The lower concentrations were found in deciduouns teeth. In both dentitions the F-content increases at the surface following the permanent change between de- and remineralization. In subsurface areas the values are decreasing quickly. At a deep of 10 microns from surface layer the content is only 70 micrograms/g and than it declines to a minimum of 20-30 micrograms/g.

Child↗

Electron probe microanalysis of secondary carious lesions associated with silver amalgam fillings.

Secondary caries associated with silver amalgam fillings is characterized by outer lesions and cavity wall lesions. In this study the content of minerals and the penetration of elements from the amalgam into such lesions were analyzed. In sections of 11 teeth including cases of natural secondary caries and experimental in vitro and in vivo lesions around silver amalgam fillings the distribution of Ca, P, Mg, Zn, Sn, Cu, Ag and Hg was studied by means of two-dimensional X-ray images, linear scans and point analyses. Dentine wall lesions where microradiographs had shown increased radiopacity relative to intact tissue, exhibited considerably reduced Ca and P values. The outer portion of the radiopaque areas contained 5--8% Zn and Sn, decreasing to less than 0.1% at a varying distance up to 130 mum from the cavity wall. Hg was not detected, nor was Ag, except in one specimen. The increased radiopacity of this zone as observed on microradiographs is, therefore, obviously caused by the presence of Zn and Sn. Zn and Sn had also penetrated into the moderately demineralized enamel lesions, but the concentrations were generally lower than those observed in the dentine.

Calcium↗

Electron probe microanalysis of secondary carious lesions adjacent to silicate fillings.

Secondary caries associated with silicate fillings is characterized by lesions at the tooth surface and lesions of the cavity wall. The mineral content of the cavity wall lesions and the penetration of elements originating from the silicate fillings were studied in experimental in vitro and in vivo lesions as well as in natural carious lesions. Dentine wall lesions, where microradiographs had shown increased radiopacity relative to intact tissue, exhibited increased Ca and P values. Elements derived from the silicate fillings were regularly found in enamel and dentine. The concentrations of fluorine (F) and of zinc (Zn) amounted to 2-3% by weight close to the cavity wall and decreased gradually to less than 0.1% at depths of 600 mum and 400 mum, respectively, from the cavity. Aluminum (Al) most often occurred in a 20-40 mum-wide zone, showing a maximum concentration of 2-3% near the cavity. Sulfur (S) was often present in the dentinal cavity walls of natural secondary caries, but not in the in vitro specimens. The findings indicate that F released from the silicate filling significantly modifies the progress of a carious lesion in the adjacent enamel and dentine. Through its tendency to form complexes with F, Al may possibly enhance the cariostatic effect of F.

Aluminum↗

Fluoride uptake around cavity walls; two-dimensional mapping by electron probe microanalysis.

This study reports fluoride uptake around the cavity wall of teeth by two-dimensional mapping. Fluoride concentration was measured using the wavelength dispersive x-ray analysis (WDX) method. The buccal cavity wall of a human tooth was coated five times with 2% sodium fluoride solution at three-day intervals for 12 days, and then immersed in a normal saline solution at 37 degrees C. After one month, the tooth was bisected longitudinally through the center of the cavity surface perpendicular to the axial wall. On the polished surface of the cut tooth, the fluoride concentration was measured. Fluoride distribution maps around the cavity wall were drawn using a bundle of the observed analytical lines. Fluoride uptake from fluoride-releasing materials (conventional glass-ionomer cement, light-cured glass-ionomer cement, light-cured composite resin, light-cured bonding agent) around the cavity wall was investigated using the same method. The maps showed higher fluoride uptake in dentin than in enamel and a strong location dependence of fluoride uptake in a tooth, especially in the dentin. Fluoride uptake from the resin was greater than that from the cement. It was summarized from these results that a two-dimensional map of fluoride uptake can provide valuable information on the cariostatic properties of fluoride-releasing materials.

Acid Etching, Dental↗

Intracellular calcium store and transport of elements in acinar cells of the salivary gland determined by electron probe X-ray microanalysis.

Electron probe X-ray microanalysis using freshly frozen hydrated and dried thin sections of dog submandibular gland was performed to determine the distribution of elements and water in the acinar cells of resting and stimulating states. The results obtained are as follows: (a) The secretory granules contained high concentrations of Ca and S while high concentrations of K and P were present in the cytoplasm and/or nucleus of acinar mucous cells of the gland in the resting state. (b) With pilocarpine stimulation, the concentration of Ca increased in the cytoplasm and decreased in the secretory granules, while there was an increase in the concentration of Na and Cl in both the cytoplasm and secretory granules of the cells. (c) The local dry-mass fractions of acinar cells, estimated by comparing the continuum radiation of X-ray spectrum from the frozen hydrated sections with that from the frozen dehydrated sections, were approximately 20 and 33% in the cytoplasm and secretory granules of resting acinar cells, respectively, and each value was not significantly altered under conditions of stimulation having a tendency to decrease slightly. Therefore, the passive Na and Cl influx and the cytoplasmic Ca flowed in from extracellular spaces and released from secretory granules, an intracellular calcium store, by secretory stimulation probably triggers the passive or active Na and Cl extrusion and consequently the osmotic water flux from the basal part of acinar cells to the secretory granules and the lumen, as well as the serial exocytosis of the granules in the luminal side of the acinar cells.

Animals↗

Three-dimensional spatial relationship between the collagen fibrils and the inorganic calcium phosphate crystals of pickerel (Americanus americanus) and herring (Clupea harengus) bone.

High-voltage (1.0 MV) electron microscopy and stereomicroscopy, electron probe microanalysis, electron diffraction and three-dimensional computer reconstruction, have been used to examine the spatial relationship between the inorganic crystals of calcium phosphate and the collagen fibrils of pickerel and herring bone. High-voltage stereo electron-micrographs were obtained of cross-sections of the cylinder-shaped intramuscular bones in uncalcified regions, in regions where only one or only several crystals had been deposited in some of the fibrils, and in successive sections containing progressively more mineral crystals until the stage of full mineralization was reached. High-resolution electron probe microanalysis confirmed that the electron-dense particles contained calcium and phosphorus. In the earliest stages of mineralization and progressing throughout the mineralization process, the crystals are located only within the collagen fibrils; crystals are not observed free in the extracellular spaces between collagen fibrils. The progressive increase in the mass of mineral deposited in the bone tissue with time occurs, essentially, completely within the collagen fibrils including the stage of full mineralization. At this stage, cross-sectional profiles of collagen fibrils are completely obliterated by mineral. A small number of crystals that are located on or close to the surface of the fibrils appear to extend a very short distance into the spaces between the fibrils. These ultrastructural observations of the very onset of calcification in which nucleation of the calcium phosphate crystals is clearly shown to begin within specific volumes of collagen fibrils, and of the subsequent temporal and spatial sequences of this phenomenon, which shows that calcification continues wholly within the collagen fibrils until maximum calcification is achieved, add important information on the basic physical chemical mechanism of the calcification and the structural elements that are involved. The spatial and temporal independence of the sites where mineralization is initiated establishes that such ultrastructural locations within individual collagen fibrils represent independent, physical chemical nucleation loci. The findings are totally inconsistent with the proposal that crystals must first be deposited in matrix vesicles, or other components such as mitochondria, and subsequently released and propagated in the interfibrillar space, until they eventually reach and impregnate the hole zone regions of the collagen fibrils. Three-dimensional computer reconstruction of serial transverse and longitudinal sections demonstrates periodic swellings along the collagen fibrils, corresponding to the hole zone region of their axial period as mineralization proceeds.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗