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Preparation of unfixed and undecalcified frozen sections of adult rat periodontal ligament during experimental tooth movement.

The upper first molars of adult male rats were moved for 7 days and unfixed, undecalcified frozen sections of the molar periodontal ligament were prepared and observed. The upper jaws of the rats were immersed rapidly in liquid nitrogen and sectioned with a cryostat using a super hard knife. Five micrometer serial sections were cut, collected, freeze-dried and observed with both light and scanning electron microscopy. Electron probe microanalysis (EPMA) was also performed on the sections. On the tension side of the periodontal ligament, periodontal fibers were stretched and the osteoblasts were aligned on the osteoid, which showed metamasia with the toluidine blue stain. On the pressure side where the periodontal ligament was extremely compressed, tissue degeneration was caused by tooth movement and the osteoclasts were observed on the bone surface adjacent to the degenerating tissues. Scanning electron microscopy revealed a network arrangement of the collagen fiber bundles on the tension side, but not on the pressure side of the periodontal ligament. The spectrum obtained from EPMA of the osteoid demonstrated X-ray (Ka) peaks of Na, P, S, K and Ca.

Animals↗

Cross-linked polypentapeptide of elastin as a calcifiable matrix: molecular weight dependence.

The polypentapeptide, (L X Val1-L X Pro2-Gly3-L X Val4-Gly5)n, when cross-linked by gamma-irradiation was shown to calcify when exposed to dialysates of calcium and phosphate augmented fetal bovine sera and the molecular weight dependence of this calcification is investigated. Five molecular weight fractions, labeled I to V in order of increasing polymer size from under 12,000 dalton (I), that is, n less than 30, to over 100,000 daltons (V), that is, n greater than 240, were gamma-irradiation cross-linked at 10-12 MRAD to form matrices I-V. Calcium-45 was used to follow the time course and relative amount of calcium uptake from the sera. Scanning electron microscopy and electron probe microanalysis were used to characterize the extent of matrix calcification. All matrices took up calcium-45 from the sera; however, only matrices formed from polypentapeptide with n greater than 100 calcified, that is, matrices III, IV, and V. Matrix V with n greater than 240 calcified massively and in a manner comparable to chemically cross-linked polypentapeptide with n approximately 40 using nonaugmented sera. Presumably, gamma-irradiation results in chain breakage. The gamma-irradiation cross-linked matrices with values of n ranging from under 30 to greater than 240 establish the molecular weight dependence of matrix calcification.

Animals↗

Hexahedrally based crystals in human tooth enamel.

Mg-containing calcium phosphate crystals including pseudocuboidal, rhombohedral shapes and groupings of quadrangular blades cubically arranged were found in human tooth enamel by scanning electron microscopy and by electron probe microanalysis. In caries-free old enamel, these hexahedrally based crystals measuring 0.5-2.5 microns in length were observed in some crevices of tufts and lamellae. The crystals were rarely seen in the inner crevices of caries-free exfoliated deciduous enamel and none could be seen in sound young enamel. In brown-coloured old enamel possessing arrested caries with lamellae, some of the lamellae contained crystals measuring 0.1-1.5 mu in length adjacent to half-dissolved prisms. These crystals, identified as Mg-containing whitlockite, will grow during a long period after eruption of the tooth or during the enamel caries process.

Aged↗

Electron microprobe analysis in periodontal guided tissue regeneration.

Electron microprobe analysis was used to determine the evolution of Ca, P and S in regenerated tissue surrounding incisors roots after periodontal treatment with guided tissue regeneration. Our results, which showed increased Ca and P, and decreased S are discussed in relation to the process of mineralization electron probe microanalysis with potentially provided an accurate means of assessing the degree of mineralization in extremely small tissue samples.

Calcium↗

Elemental analysis and clinical implications of calcification deposits associated with silicone breast implants.

Calcification of the fibrous capsule surrounding silicone breast implants is a well-recognized occurrence that increases with time following implantation. These mineralized deposits potentially confound mammographic breast cancer surveillance already made difficult by the obscuring effects of silicone breast implants. The authors performed elemental analysis of silicone breast implant-associated calcifications to define better their chemical composition as related to mammographic and clinical significance. Electron probe microanalysis and infrared spectroscopy revealed all of the calcification deposits to be calcium complexed with tribasic phosphate. No evidence of calcium oxalate, calcium carbonate, silicone, or talc was observed. Caution must be employed in interpreting mammograms in women with silicone breast implants as well as those who have had their silicone breast implants removed. High-density mammographic calcifications indicative of calcium phosphate associated with a silicone breast implant may represent an accepted consequence of implantation or nearby carcinoma. We recommend baseline mammography on women who have had their silicone breast implants removed to prevent unnecessary fine-needle aspiration or tissue biopsy of retained breast capsule calcifications during subsequent routine surveillance for carcinoma.

Breast↗

Renal sodium-potassium adenosine triphosphatase. Optical localization and x-ray microanalysis.

The distribution of sodium-potassium adenosine triposphatase (Na-K-ATPase) activity in kidney sections has been studied by a method based on the hydrolysis of p-nitrophenyl phosphate in alkaline medium containing dimethyl sulfoxide. The products at each stage in the reaction sequence have been subjected to electron probe microanalysis. The initial product was identified as a mixture of KMgPO4 and Mg(PO4)2, and sequential analysis demonstrated the linearity of conversion of this product to a visible form. In human, rabbit and rat kidneys the distribution of activity was found to be essentially identical, with highest levels located in thick ascending limbs and distal convoluted tubules. The initial reaction was completely potassium dependent and was inhibited by ouabain in concentrations reflecting the relative sensitivity of microsomal Na-K-ATPase in each species. Measurement of initial product phosphorus by means of the electron probe is presented as a practical technique for direct quantitation of Na-K-ATPase activity in identified tubule segments.

Adenosine Triphosphatases↗

Ultrastructure of selected struvite-containing urinary calculi from cats.

OBJECTIVE: To elucidate the ultrastructural details of struvite-containing urinary calculi from cats. DESIGN: Specimens studied were inclusive of the range of textures visible during preliminary analysis by use of a stereoscopic dissecting microscope. Textural types, which were used to infer crystal growth conditions, were differentiated with regard to crystal habit, crystal size, growth orientation, and primary porosity. SAMPLE POPULATION: Thirty specimens were selected from a collection of approximately 1,600 feline urinary calculi: 20 of these were composed entirely of struvite, and 10 consisted of struvite and calcium phosphate (apatite). PROCEDURE: Qualitative and quantitative analyses of specimens included use of plain and polarized light microscopy, x-ray diffractometry, scanning electron microscopy with backscattered electron imagery, x-ray fluorescence scans, and electron probe microanalysis. RESULTS: Four textural types were recognized among struvite calculi, whereas 2 textural types of struvite-apatite calculi were described. CONCLUSIONS: The presence of minute, well interconnected primary pores in struvite-containing urinary calculi from cats is an important feature, which may promote possible interaction of calculi with changes in urine composition. CLINICAL RELEVANCE: Primary porosity, which can facilitate interaction between the calculus and changing urine composition, may explain the efficacy of dietary or medicinal manipulations to promote the dissolution of struvite-containing uroliths from this species.

Animals↗

Bone-bonding behavior of three heat-treated silica gels implanted in mature rabbit bone.

Silica gel has been reported to induce apatite nucleation on its surface in vitro and it can act as a stimulant that induces formation of chemical apatite (Ca-P) layers on the surfaces of bioactive glass-ceramics. In this study, apatite formation in response to and the bone-bonding behavior of silica gels implanted in the tibiae of mature rabbits were studied. Implants were made from three silica gels treated at 400, 800, and 1000 degrees C, and the effects of such heat treatment on the above parameters were investigated. The silica gel was made by hydrolysis and polycondensation of tetraethoxysilane in aqueous solution containing polyethylene glycol. Rectangular implants (15 mm x 10 mm x 2 mm) of each heat-treated silica gel were implanted into both tibial bones of mature male rabbits, which were killed 4 or 8 weeks after implantation, and the tibiae containing the implants were dissected out. The bone-implant interfaces were investigated using Giemsa surface staining, contact microradiography, scanning electron microscopy-electron probe microanalysis, and X-ray diffraction. Histologically, no bonding of bone to any of the silica gels was observed at any time postimplantation. Soft tissue was observed at the bone-silica gel interface, but there were no giant foreign body or inflammatory cells. A Ca-P-rich layer was observed only on small areas of the surfaces of the silica gels treated at 400 and 800 degrees C 4 and 8 weeks after implantation. X-ray diffraction analysis confirmed the presence of hydroxyapatite in these Ca-P-rich layers.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Bonding of chemically treated titanium implants to bone.

A study was undertaken in rabbit tibiae to determine the effects of chemical treatments and/or surface-induced bonelike apatite on the bone-bonding ability of titanium (Ti) implants. Smooth-surfaced plates (10 x 10 x 2 mm) of pure Ti, alkalil- and heat-treated Ti, and bonelike apatite-formed Ti after the treatments were implanted into the tibial metaphyses of mature rabbits. The tibiae containing the implants were harvested at 4, 8, and 16 weeks after implantation and subjected to a tensile testing and histologic evaluation. Biomechanical results showed that both treated implants exhibited significantly higher failure loads compared with untreated Ti implants at all time periods. Histologic examination by Giemsa surface staining, contact microradiography (CMR), and scanning electron microscopy (SEM) in backscatter mode revealed that both treated Ti implants directly bonded to bone tissue during the early postimplantation period, whereas untreated Ti implants formed direct contact with the bone only at 16 weeks. SEM-electron-probe microanalysis (EPMA) examination showed a Ca-P-rich layer at the interface between the treated implants and bone, although the Ca-P-rich layer was not detected on the surface of untreated implants during observation periods. The results of this study suggest that chemical treatments may accelerate the bone-bonding behavior of titanium implants and enhance the strength of bone-implant bonding by inducing a bioactive surface layer on Ti implants.

Animals↗

[Chemical analysis of the integument and digestive gland of Crangon crangon (Linné) (Crustacea, Decapoda) by x ray spectography. Quantitative variations in the elements of calcium, phosphorus, sulfur, and magnesium during the intermolt cycle].

Electron probe microanalysis is a very suitable method for qualitative and quantitative analysis of calcium, phosphorus, sulfur and magnesium occurring within integument and digestive gland; variations related to intermolt cycle can be compared.

Animals↗

Effect of resin-modified glass ionomer cements on secondary caries.

PURPOSE: To evaluate the in vitro secondary caries inhibitory effect of two resin-modified glass ionomer cements (R-GICs). MATERIALS AND METHODS: Class V cavities were prepared at the cementoenamel junction on facial and lingual surfaces of 32 extracted upper premolars. The facial cavities were restored with a conventional glass ionomer cement (GIC) (Fuji II), while the lingual cavities were restored with either one of the R-GICs (Fuji II LC, Photac Fil, Vitremer), or a resin composite (Z-100). The margins of the restorations were subjected to a 20-day in vitro cariogenic challenge and the degree of demineralization was analyzed by microradiography. The amounts of fluoride released into distilled water from disc specimens of the materials tested were also measured for immersion time up to 161 days. RESULTS: The R-GICs and GIC's released similar cumulative amounts of fluoride over 161 days. Photac-Fil showed significantly higher amounts of fluoride release over the same period. The depth of the outer lesion and the thickness of the acid-resistant layer showed no significant difference among the R-GICs and GIC's. Moreover, the residual fluoride and calcium in the dentin adjacent to the R-GICs and GIC's were correlated with the thickness of acid-resistant layers in the dentin adjacent to the R-GICs and GIC's by electron probe microanalysis. However, the inhibitory effect of the R-GICs was not directly related to the fluoride concentrations eluted from them.

Bicuspid↗

The effect of dietary xylitol on recalcifying and newly formed cortical long bone in rats.

Thirty-six 3-week-old male Wistar rats were labeled with a single intraperitoneal tetracycline injection. Twenty-four of them were then fed a Ca-deficient basal diet for 3 weeks, while the control group received the basal diet supplemented with CaCO3 (12 g/kg). The tetracycline labeling was then repeated and six animals in each group were decapitated. The diet of the remaining formerly Ca-deficient animals was returned to normal, and half the test rats also received xylitol supplementation (50 g/kg). After 4 weeks of rehabilitation the labeling was repeated and the animals were decapitated and their tibias were prepared. The tibias were measured in terms of weight and density and cross sections were prepared for the examination of mineral content. Bone element analysis was performed by scanning electron microscopy with electron-probe microanalysis, examining separately the bone areas formed during the various dietary periods. Areas of the former Ca-deficient and newly formed cortical bone were identified by tetracycline fluorescence under ultraviolet light, and the amount of cortical bone in each group was measured. The mineralization-promoting effect of dietary xylitol as compared with CaCO3 supplementation alone was seen more clearly in the newly formed periosteal bone than in remineralization of the formerly Ca-deficient bone, the concentrations of Ca and P being significantly elevated (P < 0.05), as also was the total mineral content (P < 0.01). The cortical bone volume was similar following the CaCO3 and CaCO3 + xylitol supplementations, suggesting unaltered formation of the organic matrix. The results show that the effect of xylitol on bone during dietary Ca rehabilitation particularly concerns newly formed bone mineral.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Silicone deposition in reconstruction scars of women with silicone breast implants.

BACKGROUND: The possible association of silicone breast implants and disease is a subject of continuous debate and concern. OBJECTIVE: Our purpose was to examine microscopically and ultrastructurally the periprosthetic fibrous capsules and reconstruction scars of women with silicone breast implants. METHODS: Representative samples from the periprosthetic capsules and reconstruction scars from six women with silicone breast implants were examined by a variety of light microscopy techniques, transmission electron microscopy, and electron probe microanalysis. RESULTS: Silicone globules of various sizes were identified in every periprosthetic capsule and reconstruction scar. CONCLUSION: Extrusion and seeding of the incision tract during surgery most likely accounts for the presence of silicone in the reconstruction scar specimens. This observation suggests that the identification of silicone in the reconstruction scars of women with silicone breast implants does not necessarily implicate rupture of the silicone breast implant with systemic dissemination of silicone gel.

Breast Implants↗

Subcellular electrolyte shifts during in vitro myocardial ischemia and reperfusion.

Isolated perfused rabbit right ventricular wall was studied with electron probe microanalysis (EPMA) under three conditions: 1) control (37 degrees C, 1.2 Hz), 2) 60 min global ischemia, and 3) ischemia plus 5 min of reperfusion. After 60 min of ischemia, only one cell population was evident; the variance of intracellular electrolyte concentrations was the same as in controls. When compared with controls, there was no change in Ca concentration within any region of the cell, but mitochondria were swollen with K-rich fluid. Two cell populations were evident after 5 min of reperfusion. The severely injured cells were markedly swollen, exhibited hypercontraction bands, and had electrolyte profiles similar to extracellular fluid. The moderately injured cells were normal in appearance, still retained electrolyte gradients, but had elevated Na and Cl concentrations in all compartments. Cell Ca did not increase in the moderately injured cells, but the region of the cell containing the sarcoplasmic reticulum (SR) lost 90% of its Ca. Accompanying this loss were large increases in myofibrillar and mitochondrial Ca concentration. It appears that release of SR Ca, loss of SR Ca-accumulating capacity, and increased intracellular Na are the principal electrolyte shifts in functional cells during early reperfusion.

Animals↗

Effect of time at 37 degrees C on the creep and metallurgical characteristics of amalgam.

When aged at 37 degrees C for six months, dental amalgam exhibits a marked decrease in the property of creep. The objective of this study was to investigate the relationship between this decrease in creep and selected metallurgical characteristics. The formation of beta 1 (Ag-Hg), the grain size of gamma 1 (Ag-Hg), and the composition of gamma 1 were chosen for investigation. Creep was determined according to ADA Specification test No. 1, beta 1 was measured by x-ray diffraction, and gamma 1 grain size and gamma 1 composition were determined by electron-probe microanalysis. The results showed that the decrease in creep was related to beta 1 formation and not to changes in either gamma 1 grain size or gamma 1 composition.

Chemical Phenomena↗

[Effect of morphological properties on drug release from biodegradable microspheres].

The morphological properties of poly(beta-hydroxybutyric acid) (PHB) or poly(L-lactic acid) microspheres loading flomoxef sodium (FMOX) were investigated with regard to FMOX release. The release profiles of FMOX from the microspheres could be divided into two types, a sustained release type and a burst one. Two representative PHB microspheres, the release profiles of which were quite different from those of FMOX, were compared in detail from a morphological point of view. The shapes of their surfaces and sections were observed by using scanning electron microscopy (SEM), and FMOX distribution was analyzed by using electron probe microanalysis. The crystallinity of polymers was further measured by powder X-ray diffratometry. There was little difference in the FMOX distribution and their microscopic properties such as sphere size, specific surface area, shape of surface and section. In contrast, water penetration into the inside of the microspheres was found to be clearly different by use of cryogenic SEM. A significant difference was also observed in the crystallinity of polymers forming the microspheres. The release of FMOX from the microspheres was affected by the crystallinity of polymers forming the microspheres, and burst phenomena occurred in case the polymer was highly crystallized. It was speculated that the crystallization of polymer induced micro voids in the microspheres which functioned as channels for water penetration.

Biodegradation, Environmental↗

Changes in Schwann cells and vessels in lead neuropathy.

Transmission electron microscopy (TEM) of peripheral nerve in rats receiving 6% lead carbonate for 4-10 weeks provided evidence of a specific Schwann cell injury, associated with demyelination. Intranuclear inclusions in Schwann cells appeared within 2 weeks of administration of a lead-containing diet. Swelling of Schwann cells and disintegration of their cytoplasm was evident at 4 weeks. Distinctive electron-dense inclusions appeared in both Schwann and endothelial cells during the period of intoxication and were ultrastructurally identical to pathognomonic inclusions of lead poisoning seen in renal tubular epithelial cells. Scanning microscopy (SEM) with electron-probe microanalysis was used to identify the lead-containing deposits. In addition to Schwann cell changes, vessels revealed endothelial cell injury and alteread permeability to macromolecules. Since morphologic changes of Schwann cells precede the development of altered vascular permeability and endoneurial edema, it appears that lead gains access to the endoneurium prior to the development of altered vascular permeability, suggesting that edema and altered endoneurial fluid pressure are epiphenomena that supervene after demyelination occurs. Remyelination, Schwann cell proliferation and formation of onion bulbs are manifestations of persistent toxic injury to myelin-sustaining cells, resulting in chronic demyelination.

Animals↗

The distribution of intracellular ions in the avian salt gland.

To investigate the mechanism of salt secretion in the avian salt gland, we used quantitative electron probe microanalysis to measure the intracellular elemental concentrations in dry cryosections of unspecialized and partially specialized secretory epithelial cells from fresh water- and salt water-adapted ducklings, respectively. In conjunction with this, human and duckling erythrocytes were also analyzed, since these provided the experimental basis for using in situ erythrocytes as standards for determining the local water content of epithelia from the analysis of dried cryosections. The microprobe results from both types of erythrocytes compared favorably with chemical determinations of elemental concentrations. The nucleated avian erythrocytes, whose wet-weight elemental concentrations were determined by a compartmental analysis that required neither a peripheral standard nor a measure of the local mass, revealed a marked accumulation of P and K in the nucleus (388 and 190 mmol/kg wet wt, respectively) relative to the cytoplasm (67 and 85 mmol/kg wet wt). In both developmental states of the epithelial cells, the nucleus and apical cytoplasm had essentially similar and unremarkable concentrations of Na (76 and 83 mmol/kg dry wt, respectively, in the adapted cells vs. 72 and 81 mmol/kg dry wt in the control cells) and K (602 and 423 mmol/kg dry wt vs. 451 and 442 mmol/kg dry wt). Chloride, however, which was in general rather high, was significantly depressed in the apical cytoplasm of adapted cells only (164 and 124 mmol/kg dry wt in the nucleus and cytoplasm, respectively, of adapted cells (P less than 0.05) vs. 138 and 157 mmol/kg dry wt for control cells (P less than 0.05). Cation concentrations (Na + K) were elevated approximately 15% in the basal regions of adapted cells as compared with apical cytoplasm. When tissue water variations are accounted for, the results suggest that: (a) an active, energy-requiring process is responsible for chloride accumulation in this cell; (b) the apical membrane is a regulatory site for secretion; and (c) there are regional distinctions in the distribution of ions and water, particularly in the salt water-adapted cell. These conclusions are consistent with active chloride transport as the basis for salt secretion in this tissue.

Animals↗