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A medicated polycarboxylate cement to prevent complications in composite resin therapy.

Preparative treatment is the preferred method to protect the dentin and pulp from complications in composite resin therapy. This study investigated the in vivo effects of the polycarboxylate cement containing zinc fluoride and tannic acid in composite resin restorations. Scanning electron micrographs established that the composite resin failed to contact the axial wall. The gaps varied from 10 to 60 microns. However, this polycarboxylate cement was shown to provide excellent adaptation to dentin when used as a base and its chemical adhesion allowed it to make close contact with the unetched dentin. The newly developed electron probe x-ray microanalyzer revealed that the in vivo penetration of fluoride and zinc occurred through the dentinal tubules. When this polycarboxylate cement was used, the orifices of dentinal tubules were partially occluded, possibly with the smear layer fixed by tannic acid. In addition, by releasing the components, this polycarboxylate cement adds acid resistance to dentin and increases the resistance of dentin collagen to proteolytic enzymes. As such this polycarboxylate cement offers advantages as a base to composite resin therapy.

Adolescent↗

Analysis of vaterite microspherolith deposits on a pure cholesterol gallstone by X-ray diffraction, X-ray microanalysis and infrared absorption techniques.

Minute globular concretions of light-green and light-brown colour were found as deposits in the pits at the rough surface of a pure cholesterol gallstone. They were analyzed by scanning electron microscopic, X-ray diffraction, X-ray microanalytical and infrared absorption spectrometric methods. In scanning electron microscopy, the concretions appeared as ovoid microspheroliths varying from 5 to 30 micron in size. The microspheroliths presented smooth and rough surfaces; the reason for these different types of surface remained unclear. X-ray diffraction and infrared absorption analysis revealed that the microspheroliths were mainly composed of vaterite and the host plate-like crystals were cholesterol. By energy-dispersive X-ray microanalysis, large quantities of calcium without significant quantities of phosphorus were detected in the microspheroliths. Calcium was absent in the plate-like crystals and could not be detected in the central part of the stone. In the pigmented periphery of the stone, some silicon and iron were found. The presence of calcite, aragonite and iron-containing pigment materials in the concretion is suggested. From the textural point of view, the analytical data strongly suggest that the stone-forming conditions suddenly changed from a cholesterol-favourable stasis condition to a condition favoring the deposition of calcium carbonate possible as the result of haemorrhage.

Absorption↗

The localization of histochemical and autoradiographic products in the scanning electron microscope by means of atomic number contrast.

The role of atomic number contrast and backscattered imaging in the localization of histochemical products and reagents is reviewed. Particular attention is given to the localization of enzymatically released products. A backscattered imaging method is presented which permits the simultaneous demonstration of two label elements in a cryosection examined in the scanning electron microscope. An azo-dye method for the localization of acid phosphatase is combined with an autoradiographic technique for demonstrating [3H]thymidine incorporation. Atomic number contrast, produced by backscattered electrons, permits the detection of a bromine-labelled azo dye, denoting sites of acid phosphatase activity within a cryosection and the simultaneous display of the sites of the silver deposited in an overlying photographic emulsion. The former reveals acid phosphatase to be predominantly associated with thymocyte death and macrophage activity in mouse thymus. The latter pin-points the thymocytes as the main sites of [3H]thymidine incorporation. Atomic number contrast has a useful potential for differentiating between spatially separated histochemical products of differing atomic number in a structural context and also allows a flexible degree of subsurface localization.

Animals↗

Heavy metal cytotoxicity studied by electron probe X-ray microanalysis of cultured rat hepatocytes.

Cytotoxicity of the heavy metals gold, mercury, thallium and lead was studied by measuring the intracellular element distribution of cultured rat hepatocytes by energy dispersive electron probe X-ray microanalysis of freeze-dried cryosections in a scanning transmission electron microscope. Exposure of the cells to aqueous solutions containing heavy metal ions in concentrations reaching a critical concentration caused increase of intracellular sodium and chloride content accompanied or followed by decrease of intracellular potassium content. Thus, the intracellular potassium/sodium ratio drastically decreased from control values of approximately 10 to values below 1 before changes of cell morphology became visible. In experiments with gold or mercury the decrease of the potassium/sodium ratio was preceded by transient cytoplasmic increase of sulfur and phosphorus. Heavy metal concentrations exceeding the critical concentration also caused an increase of cytoplasmic calcium concentration and finally decay of the cell structure. Cytotoxicity of heavy metals was found to increase in the order Pb, Au, Tl, Hg. Cytotoxic effects by Au, Tl or Hg in moderate concentrations were reduced by simultaneous addition of Zn or Pb to the culture medium. The results obtained prove electron probe X-ray microanalysis of cryosections as a sensitive probe of cell viability.

Animals↗

Effect of Nd:YAG, Ho:YAG, Er:YAG, CO2, and GaAIAs laser irradiation on surface properties of endosseous dental implants.

PURPOSE: To analyze potential surface alterations in endosseous dental implants induced by irradiation with common dental lasers. MATERIALS AND METHODS: Sandblasted and acid-etched, plasma-sprayed, hydroxyapatite-coated, and smooth titanium discs were irradiated using Nd:YAG, Ho:YAG, Er:YAG, CO2, and GaAIAs lasers at various power settings. The specimens were examined by scanning electron microscopy and energy dispersive spectroscopy. RESULTS: In an energy-dependent manner, the pulsed YAG lasers induced partial melting, cracking, and crater formation on all 4 surfaces. Within the energy range applied, the CO2 laser caused surface alterations on the hydroxyapatite and plasma coatings as well as in the acid-etched surface. GaAIAs laser irradiation did not damage any of the surfaces. Energy dispersive spectroscopy revealed an altered chemical compound of the surfaces with regard to titanium, oxygen, and silicon. DISCUSSION: The clinical application of most common dental laser systems can induce implant surface alterations. Relevant factors are not only the laser system and power setting, but also the application system. CONCLUSION: The results of the study indicate that Nd:YAG and Ho:YAG lasers are not suitable for use in decontamination of implant surfaces, irrespective of the power output. With the Er:YAG and CO2 laser, the power output must be limited so as to avoid surface damage. The GaAIAs laser seems to be safe as far as possible surface alterations are concerned.

Carbon Dioxide↗

Fluoride penetration into the hybrid layer from a dentin adhesive.

PURPOSE: To evaluate the in vitro microleakage of a new fluoride-containing dentin adhesive and to provide evidence for fluoride release from the adhesive and penetration into the dentin. MATERIALS AND METHODS: Class V preparations with margins in enamel and dentin were made in the buccal and lingual surfaces of six extracted human third molars. Six preparations were restored with Scotchbond Multipurpose (SBMP)/Z100 composite and six with a new fluoride containing dentin adhesive (FB)/Litefil composite. After aging for 30 days in water at 37 degrees C, the teeth were stained with silver nitrate, sectioned and graded by two observers for leakage at the enamel and dentin margins. Selected FB specimens were examined in the scanning electron microprobe using WDS for fluoride and EDS for calcium and phosphorous. Fluoride release into water from disks of FB adhesive was evaluated for up to 112 days using a specific ion electrode. RESULTS: Fluoride leached from the adhesive at a decreasing rate with time. The final rate over 112 days was 0.02 ppm/day (0.2 microgram/cm2/day). Leakage at the enamel and dentin margins was similar for both adhesives (Mann-Whitney U-test; P < or = 0.05). SEM evaluation showed the presence of a discontinuous hybrid layer of 2-3 microns. Fluoride was present within, but limited to, the hybrid layer for the FB adhesive. The penetration of fluoride could only be confirmed in areas where leakage had occurred.

Calcium↗

The deposition of talc in patients with silicone gel-filled breast implants.

This study was initiated to understand why talc was found in the entrance wounds and pericapsular scars of patients with silicone breast implants. Twenty-five surgical gloves made between 1977 and 1992 by a major glove manufacturer were evaluated with infrared spectroscopy. Seven gloves manufactured before early 1983 contained talc. Later manufactured gloves contained calcium carbonate instead of talc. Talc-containing gloves were submitted to independent laboratories for scanning electron microscopic examination and x-ray microanalysis to obtain finer detail. Remnants of the mold-release agent talc were found in the matrix of the patient-contact side of the glove. Wetting studies showed that silicone oil (gel bleed) immediately wet the surface of rubber glove. After donning, silicone oil was massaged between the thumb and index finger of each washed, talc-containing glove for 1 minute and then daubed from the glove to a microscopic slide. The slide was viewed with polarized light microscopy for the presence of talc crystals. Transfer of talc occurred using silicone oil, but not saline, in each talc-containing glove. Three factors were likely involved in the greater talc deposition that occurred with patients undergoing silicone gel-filled breast implantation than in patients undergoing other surgical procedures. (1) The wetting of the gloves with silicone gel bleed from implants loosened the talc in the glove matrix. (2) Hand-intensive surgical use freed the talc from the gloves. (3) Loose talc adhered to the silicone elastomer of the breast implants and was deposited with implant placement.

Breast Implantation↗

Comparison of shear and flexural bond strength tests versus failure modes of dentin bonding systems.

PURPOSE: To (1) measure the shear and flexural bond strengths of three different bonding agents that use different application techniques, (2) determine whether the shear and flexural bond tests rank the three materials similarly, and (3) determine whether the fractured surface produced with the flexural test, facilitates studies of failures within the adhesive interphase region. MATERIALS AND METHODS: Scotchbond MP (SBMP), Single Bond (SNGB) and Clearfil SE (CLSE) were evaluated. For each material, 16 samples were bonded. Eight of these samples were bonded for shear testing and the other 8 samples for flexural bond strength testing. After the bonding was completed, all samples were stored in water at 37 degrees C for 30 days. Shear bond strength was measured using an orthodontic edge wire loop and flexural strength was measured using a 4-point bending device. The samples used for the flexural test consisted of 3 mm x 3 mm x 20 mm beams in which center a 1 mm thick dentin wafer had been bonded perpendicularly to the length of the beam. An indentation with a microhardness tester was placed at one of the dentin-composite interfaces to serve as an induced flaw. This flaw was placed on the surface in tension during flexural testing. The data were analyzed using ANOVA. Scanning electron microscopy was used to examine each specimen and assign its failure mode. Percent occurrence of failure mode was determined for each material and overall for both test methods. When there was uncertainty regarding failure location, electron dispersive spectroscopy x-ray analysis was used to identify elements present on the fractured surface. RESULTS: No significant difference (P > 0.05) in bond strength was observed within each test group, while significant differences (P < 0.05) existed between the two test methods (shear: SBMP = 21.2 +/- 4.0 MPa, SNGB= 24.3 +/- 4.7 MPa, CLSE= 24.6 +/- 4.4 MPa; flexural strength: SBMP= 34.6 +/- 9.3 MPa, SNGB = 31.9 +/- 6.9 MPa, CLSE= 34.3 +/- 4.7 MPa). Shear bond test specimens failed mostly within dentin (54.2%), followed by failures within the adhesive interphase (41.6%), and failures in composite (4.2%). Flexural strength specimens failed mostly within the adhesive interphase (83.3%), followed by failure in composite (16.7%). Bond strengths were similar for all three adhesive systems within each test method group. Failure mode analysis revealed significant differences (P < 0.0001) among the two test methods.

Adhesives↗

Structural and microanalytical changes in dentition after radiotherapy applied in cases of tumour in the oral cavity region.

Morphological observations by means of the scanning electron and optical microscopes as well as microanalytical qualitative and quantitative tests concerning content and distribution of calcium, phosphorus and magnesium in the teeth being in irradiation field were made in patients treated for larynx cancer by methods of radiotherapy. The teeth enamel was in some places thinner and in some others it was lacking. The characteristic defects took place in the form of notches, cracks and of superficial destruction. In X-ray microanalyser the enamel showed some features of demineralization. In dentine, features of demineralization and on the other hand features of remineralization were seen. The root dentin, in which destructive processes but also reparation processes developed by root cement could be seen, was rather characteristic. Clear structural changes and disturbances of mineralization in hard tooth tissues being under therapeutic X-ray irradiation were observed.

Adult↗

The ultrastructure of the bone-hydroxyapatite interface in vitro.

Rat bone marrow cells were cultured on plasma-sprayed hydroxyapatite (HA). The cells formed a mineralized extracellular matrix (ECM) that exhibited several characteristics of bone tissue. The interface between this mineralized ECM and the HA was studied at the ultrastructural level with scanning and transmission electron microscopy and x-ray microanalysis. Initially, the deposition of a globular, afibrillar matrix was observed on HA. This was followed by the integration of collagen fibers in this matrix and their subsequent mineralization. At the bone-HA interface two distinctly different interfacial structures were observed. An electron-dense layer with a thickness of 20-60 nm was regularly present, which contained both organic and inorganic material and was rich in glycosaminoglycans. The interfaces differed however, in the presence or absence of an amorphous zone which was free of collagen fibers and had an average thickness of 0.7-0.8 microns. It was frequently seen interposed between the electron-dense layer and the hydroxyapatite. Similar interfacial structures have also been described in the in vivo environment, where they were referred to as lamina limitans-like or cement linelike. From the results of this study, it can be concluded that the described in vitro system is a suitable model to study bone-biomaterial interactions.

Animals↗

Exogenous particles in lymph nodes in patients with shrinking pleuritis with atelectasis.

Shrinking pleuritis with atelectasis is a disease in which a compression atelectasis of the lung is caused by shrinking of the inflamed visceral pleura. This reaction has been attributed to asbestos. By means of scanning electron microscopy, with computerised x ray spectrophotometry and x ray diffraction analysis, tissue samples from the lymph nodes of patients with the disease and control subjects were examined for other particles of possible aetiological importance. The results would suggest, however, that asbestos is the most important factor in the aetiology of shrinking pleuritis with atelectasis.

Asbestos↗

Nutrient amounts of ectomycorrhizae analysed by EDX using ESEM and ICP.

Energy-dispersive X-ray (EDX) analysis coupled with an environmental scanning electron microscope (ESEM) was tested as an alternative to the inductively coupled argon plasma (ICP) spectrometer method for nutrient analyses of ectomycorrhizae. The results of EDX-ESEM and ICP were compared for 12 ectomycorrhizal morphotypes collected in beech and Scots pine forests in northern Brandenburg. The amounts of Al, Ca, Mg and S analysed in the outer hyphal layers of the sheath with the EDX-ESEM technique correlated well with the amounts of these elements in the whole mycorrhiza as assessed by ICP. For the elements P and K, no such correlation existed, indicating an uneven distribution of these elements in the ectomycorrhiza. It is concluded that the EDX-ESEM technique could be a useful and reliable tool for the analysis of nutrient elements in ectomycorrhizae, especially for studies focussing on small-scale soil heterogeneity or on infrequent morphotypes.

Electron Probe Microanalysis↗

Scanning transmission ion microscopy as it complements particle induced x-ray emission microanalysis.

The early uses of Scanning Transmission Ion Microscopy (STIM) with MeV ions are reviewed. The transformation of STIM energy-loss images into maps of areal density is discussed, and is illustrated with images of a fruit fly head (Drosophila melanogaster). Freeze-dried male heads are transparent to 4-MeV protons in the dorsal and frontal directions, but in the sagittal direction the brain is opaque. STIM with molecular ions is shown to be useful for increasing contrast in low density areas. For recording registered STIM and PIXE images without changing accelerator parameters, apertures are used to accomplish the required change in beam intensity (a factor of 10(5]. Molecular ions are used to assess contamination of the microbeam by scattered ions. Pixel by pixel ratios of x-ray intensity to areal density are taken to obtain maps of element concentrations. Calcium and iron maps are shown. Inner parts of the fly head are clearly seen in the concentration maps. The PIXE exposure caused differential displacements of inner parts of the head by 15 micron or less. Weight loss during the PIXE exposure was measured to be 3%.

Animals↗

Probe size and bound label conformation in colloidal gold-ligand labels and gold-immunolabels.

Colloidal gold can be produced in sizes ranging from 1.0nm to 150nm. All sizes of gold can be conjugated, principally by hydrophobic bonding, to a variety of molecules including ligands, enzymes and antibodies, as well as lectins and polysaccharides. The activity of most of these biological molecules is retained on conjugation with gold particles irregardless of size range, although the ratio of protein surface area to gold particle surface area varies widely depending on particle and protein size. We have employed low voltage high resolution scanning electron microscopy to compare, microscopically, the shapes of biological molecules unbound, bound to very small (3nm) gold particles, and bound to larger (18nm-30nm) gold particles. When very small gold particles are conjugated to large protein molecules, several particles bind along the length of each molecule, while smaller protein molecules often wrap around a single small gold particle. With larger gold particles, several biological molecules bind to a single gold particle. In addition, the shape of protein molecules bound to larger gold particles differs from that of molecules bound to small gold particles.

Animals↗

Calcium associated with a fibrillar organic matrix in the scleractinian coral Galaxea fascicularis.

Field emission scanning electron microscopy of frozen-hydrated preparations of the scleractinian coral Galaxea fascicularis revealed organic fibrils which have a diameter of 26 nm and are located between calicoblastic ectodermal cells and the underlying CaCO(3) skeleton. Small (37 nm in diameter) nodular structures observed upon this fibrillar organic material possibly correspond to localised Ca-rich regions detected throughout the calcifying interfacial region of freeze-substituted preparations by X-ray microanalysis. We propose that these Ca-rich regions associated with the organic material are nascent crystals of CaCO(3). Significant amounts of S were also detected throughout the calcifying interfacial region, further verifying the likely presence of organic material. However, the bulk of this S is unlikely to be derived from mucocytes within the calicoblastic ectoderm. It is suggested that in the scleractinian coral G. fascicularis, nodular crystals of CaCO(3) establish upon a fibrillar, S-containing, organic matrix within small but distinct extracellular pockets formed between calicoblastic ectodermal cells and skeleton. This arrangement conforms with the criteria necessary for biomineralisation and with the long-held theory that organic matrices may act as templates for crystal formation and growth in biological mineralising systems.

Animals↗

Large-scale fabrication of protein nanoarrays based on nanosphere lithography.

Hexagonally patterned lysozyme nanoarrays have been assembled on silicon wafers by combining nanosphere lithography and surface silane chemistry using vapor and solution deposition processes. The patterned protein regions extend over cm sized regions, and the size of each island is approximately 120 nm for the solution-prepared template and approximately 60 nm for the vapor-prepared template. Antibody test indicates that the patterned lysozyme maintains its bioactivity on the surface. This new approach offers a fast and reliable method to fabricate protein arrays over large areas with feature sizes comparable to scanning-probe based techniques.

Biosensing Techniques↗

Ameloblast modulation and changes in the Ca, P, and S content of developing enamel matrix as revealed by SEM-EDX.

Freeze-dried rat incisors were examined by high-resolution scanning electron microscopy (SEM) combined with energy-dispersive x-ray microanalysis (EDX) for determination of the correlation between the morphology of the enamel organ and the concentrations in the adjacent developing enamel matrix of calcium (Ca), phosphorus (P), and sulfur (S), as well as the Ca/P ratio. In SEM examination of the freeze-dried enamel organ, it was possible to identify the stages of enamel secretion, transition, and maturation, and furthermore to identify ruffle-ended and smooth-ended maturation ameloblasts. EDX analysis of the outer layer of forming and maturing enamel was carried out from the apical to the incisal end at interval points of approximately 50 micron. Ca and P concentrations increased gradually and continuously from the secretion zone to the end of the maturation zone, but never showed a steep rise in any of the zones examined. Maturing enamel overlaid by either ruffle-ended or smooth-ended maturation ameloblasts showed similar Ca and P concentrations. Throughout the outer enamel layer, the Ca/P molar ratio was fairly constant. Sulfur concentration began to decrease in the zone of enamel secretion, and was no longer detected in the middle of the maturation zone.

Ameloblasts↗

The role of bacteria in gallbladder and common duct stone formation.

Debate continues as to the role that bacteria play in gallstone pathogenesis in Western countries. We therefore, examined gallbladder and common duct stones from 67 consecutive patients undergoing cholecystectomy and/or common bile duct exploration. Bile was cultured and stone cholesterol content was measured. Stones were examined by scanning electron microscopy (SEM) for bacteria. Individual calcium salts were classified by windowless energy-dispersive x-ray microanalysis. Gallbladder stones in 65 patients were identified as cholesterol in 46 (71%), black pigment in 17 (26%), and brown pigment in 2 patients (3%). Common bile duct stones from ten patients were cholesterol in 4, black pigment in 2, and brown pigment in 4 patients. The five patients with brown pigment stones were significantly (p less than 0.05) older, more likely to be men and to present with bile duct obstruction. Bile cultures were positive in 13% of patients with cholesterol stones, in 14% of those with black pigment stones, and in all of the patients with brown pigment stones (p less than 0.001). By SEM, bacteria were observed only within the calcium bilirubinate-protein matrix of brown pigment stones (p less than 0.001). In comparison to black pigment stones, brown stones were more likely to contain calcium palmitate (p less than 0.005) and cholesterol (p less than 0.001). We conclude that black and brown pigment stones have different pathogenic mechanisms and that bacterial infection is important only in the formation of brown pigment stones.

Adult↗

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