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PubMed1989

Abrasives in snuff?

The purpose of this study was to determine and calculate the inorganic contents of four brands of snuff. Visual inspection of wet snuff showed fairly large, yellow crystal-like particles. Scanning electron microscopy and X-ray dispersive (EDX) analyses were used to study both wet snuff and ashes of snuff, whereas light emission spectrography was used to determine elements in the ashes. The crystal-like particles did not dissolve in distilled water or in ethanol heated to 60 degrees C. EDX analyses showed that most elements remained in the particles after washing. The total weight percentage of inorganic material in snuff was calculated after burning dried snuff until constant weight was obtained. The ashes of snuff did not contain any crystal-like particles but consisted of a small-grained amorphous mass. The following elements were detected: Ag, Al, Ba, Ca, Co, Cr, Cu, Fe, K, Li, Mg, Mn, Na, P, Pb, Si, Sr, Ti, Va, and Zr. Other elements such as rare earths were not searched for. The weight percentage of inorganic elements ranged between 12.35 +/- 0.69 and 20.95 +/- 0.81. Provided snuff is used in the same manner as chewing tobacco, and some people admit to doing so, there is a risk that its relatively high contents of inorganic material and heavily soluble salts may be conducive to excessive abrasion of teeth and restorations.

Calcium↗
PubMed1986

Correlated observations and analysis of maturation-ameloblast morphology and enamel mineralization.

A combined HCl-collagenase digestion technique and scanning electron microscopy were used to isolate the enamel organ and to confirm the presence of maturation ameloblasts of both ruffle-ended (RA) and smooth-ended (SA) types on maturing enamel in kitten permanent tooth germs. EDTA perfusion of animals fixed with aldehyde produced two or three belt-like shallow grooves (from 30 to 100 micron wide) running horizontally through the maturing enamel surface, coinciding closely with the SA distribution pattern. In animals that had been perfusion-fixed with unbuffered osmium tetroxide containing 2.5% potassium pyroantimonate, SEM-EDX analysis detected K in a superficial enamel layer overlaid by the SA layer. Potassium concentration decreased gradually toward the deeper layers. Very little K penetrated the enamel under the RA layer. Energy-dispersive x-ray analysis of Ca and P concentrations in the enamel revealed an even distribution of these elements throughout the superficial layer of maturing enamel. These results suggest that the SA layer forms an access route for K and EDTA and that, in spite of the obvious morphological and functional differences between RA and SA, the maturing enamel surfaces overlaid by these two cell types show similar degrees of mineralization.

Ameloblasts↗
PubMed1986

Scanning electron microscope and electron microprobe study of reactions of stannous fluoride and stannous chloride with dental enamel.

Stannous fluoride but not stannous chloride has shown a caries inhibiting effect in animal studies. The effect of aqueous solutions of stannous fluoride and stannous chloride on dental enamel surfaces was investigated by scanning electron microscope and analytical electron microprobe. Some large globules and a continuous layer of small globules were observed after stannous fluoride treatment. The large globules were soluble in both water and alkali, whereas the small ones were only soluble in alkali. Both fluoride and tin were found to be present on the enamel surfaces by the electron microprobe after stannous fluoride treatment. The fluoride could not be detected after immersion in distilled water whereas alkali treatment removed the tin. The large fluoride-containing globules may consist of calcium fluoride as this substance is known to be slightly soluble in water and soluble in alkali. Stannous chloride treatment gave a layer of small tin-containing globules with properties similar to the layer formed on enamel treated with stannous fluoride. It is therefore suggested that the tin-containing layer is not associated with caries protection whereas calcium fluoride-like deposition on the enamel probably is a factor in the caries inhibition caused by stannous fluoride.

Dental Enamel↗
PubMed1994

Bioactive glass-ceramic containing crystalline apatite and wollastonite initiates biomineralization in bone cell cultures.

Rat bone cells were cultured in the presence of bioactive glass-ceramic containing crystalline apatite and wollastonite. Scanning electron microscopy observations of the surface of the seeded ceramic disks revealed that cells attached, spread, and proliferated on the material surface. Soaking in cell-free culture medium showed that no change occurred in the surface structure. However, when cultured with bone cells and observed under a transmission electron microscope, an electron-dense layer was noted initially at the surface of the material, before bone formation occurred. In addition, energy-dispersive X-ray microanalysis demonstrated the presence of calcium and phosphorus in this layer. Progressively, during the following days of culture, active osteoblasts synthetized and laid down an osteoid matrix composed of numerous collagen fibrils arranged either parallel or perpendicularly to the first-formed electron-dense layer. Mineralization initiated on the ceramic surface dispersed then along the collagenous fibrils, leading to a mineralized matrix which surrounded the ceramic particles. These results demonstrate the capacity of apatite-wollastonite glass ceramic to initiate biomineralization in osteoblast cultures and to achieve a direct bond between the surface apatite layer of the bioactive glass-ceramic and the mineralized bone matrix.

Animals↗
PubMed1999

Effects of static or dynamic mechanical stresses on osteoblast phenotype expression in three-dimensional contractile collagen gels.

Studies performed at tissular (three-dimensional, 3-D) or cellular (two-dimensional, 2-D) levels showed that the loading pattern plays a crucial role in the osteoblastic physiology. In this study, we attempted to investigate the response of a 3-D osteoblastic culture submitted to either no external stress or static or dynamic stresses. Rat osteosarcoma cells (ROS 17/2.8) were embedded within collagen type I lattices and studied for 3 weeks. Entrapment and proliferation of cells within the hydrated collagen gel resulted in the generation of contractile forces, which led to contraction of the collagen gel. We used this ability to evaluate the influence of three modes of mechanical stresses on the cell proliferation and differentiation: (1) the freely retracted gels (FRG) were floating in the medium, (2) the tense gels (TG) were stretched statically and isometrically, with contraction prevented in the longitudinal axis, and (3) the dynamic gels (DG) were floating gels submitted to periodic stresses (50 or 25 rpm frequency). Gels showed maximum contraction at day 12 in 50 rpm DG, followed by 25 rpm DG, then FRG (88%, 81%, 70%, respectively) and at day 16 in TG (33%). The proliferation rate was greater in TG than in FRG (+52%) but remained low in both DGs. Gel dimensions were related to the collagen concentration and on a minor extent to cell number. Cells in DG appeared rounder and larger than in other conditions. In TG, cells were elongated and oriented primarily along the tension axis. Scanning electron microscopy (SEM) showed that tension exerted by cells in TG led to reorientation of collagen fibers which, in turn, determined the spatial orientation and morphology of the cells. Transmission electron microscopy (TEM) performed at maximum proliferation showed a vast majority of cells with a distended well-developed RER filled with granular material and numerous mitochondria. Alkaline phosphatase activity peaked close to the proliferation peak in FRG, whereas in TG, a biphasic curve was observed with a small peak at day 4 and the main peak at day 16. In DG, this activity was lower than in the two other conditions. A similar time course was observed for alkaline phosphatase gene expression as assessed by Northern blots. Regardless of the conditions, osteocalcin level showed a triphasic pattern: a first increase at day 2, followed by a decrease from day 4 to 14, and a second increase above initial values at day 18. Microanalysis-x indicated that mineralization occurred after 14 days and TEM showed crystals within the matrix. We showed that static and dynamic mechanical stresses, in concert with 3-D collagen matrices, played a significant role on the phenotypic modulation of osteoblast-like cells. This experimental model provided a tool to investigate the significance and the mechanisms of mechanical activity of the 3-D cultured osteoblast-like cells.

Alkaline Phosphatase↗
PubMed2003

A palladium-free gallium-based alloy: analysis of composition and microstructure.

OBJECTIVES: To study the microstructure of a palladium-free gallium-based alloy (Galloy) before and after mixing to identify the metallic phases produced by the setting reaction and assess the quantitative elemental analysis of the phases present. METHODS: The Galloy powder was analyzed using X-ray diffraction (XRD) and scanning electron microscopy (SEM) and the particle size distribution was determined using a high resolution particle size analyzer. Cylindrical test specimens were mechanically condensed in accordance with ISO 1559: 1986 for XRD, energy dispersive spectroscopy (EDS) and SEM analyses of the set alloy. Disc shaped specimens were prepared by manual condensation to compare the width of the reaction zone with specimens condensed mechanically. RESULTS: The Galloy powder consisted of spherical particles varying between 0.1 and 40 microm in diameter which contained a network of grooves and clefts extending deep within the substance of the particles. The reaction zone appeared 'grainy' and 'fragmented' compared with the well defined reaction zone previously observed for Pd-containing gallium-based alloys. The student t-test indicated the width of reaction zone varied significantly (P<0.001) with method of specimen fabrication.Significance. The significantly larger reaction zone in the manually condensed specimens was possibly due to failure of the condensation technique to force out excessive liquid alloy present in the triturated mass. The surface topography of the Pd-free Galloy powder particles could be responsible for the characteristically fragmented and ill-defined reaction zone observed in the set material compared with the well defined reaction zone previously observed for Pd-containing gallium-based alloys.

Dental Alloys↗
PubMed2003

Occluding effect of dentifrices on dentinal tubules.

OBJECTIVES: The objective of this study was to evaluate the tubule occluding ability of three commercial available dentifrices (Sensodyne, Emoform and Sorriso) by Scanning Electron Microscopy. METHODS: Fifty cervical areas from buccal and lingual surfaces of sound third human molars were used. Cervical enamel was wet abraded to expose flat dentin surfaces and further polished with diamond pastes. Specimens were randomly divided into five groups (n=10): G1-no brushing; G2-brushing without dentifrice; G3-brushing with Sensodyne; G4-brushing with Emoform; G5-brushing with Sorriso. Brushed specimens were treated for 4 min per day, for 7 days in a toothbrushing machine. Specimens were prepared and observed under SEM for calculation of the percentage of occluded tubules. In addition, slurries of toothpastes were analyzed by X-ray microanalysis. Data were statistically analyzed by ANOVA and Tukey test (p<0.05). RESULTS: Means of occluded tubules in descending order were: G5-98.83+/-3.31% (a), G4-96.02+/-5.24% (a), G3-80.12+/-24.65% (a), G1-37.31+/-24.22% (b); G2-22.92+/-15.19% (b). The three tested dentifrices produced increased dentinal occlusion as compared to controls (p<0.05) but equivalent occlusion among each other. Calcium from calcium-carbonate abrasive was identified in all dentifrices. CONCLUSIONS: Results indicated that the use of all dentifrices occluded tubules more than no brushing and brushing without dentifrices groups. Thus, the tested dentifrices seem effective for desensitization by tubule occlusion.

Analysis of Variance↗
PubMed1987

Automated gunshot residue particle search and characterization.

The main disadvantage to gunshot residue (GSR) particle analysis utilizing scanning electron microscope/energy dispersive X-ray (SEM/EDX) instrumentation has been the excessive operator time required for search and identification. This study uses an automated particle search and characterization program for unattended GSR search and identification. This system allows for automatic matrix search, particle sizing, chemical typing, and spectral aquisition with subsequent storage of data to disk for later operator review and verification. This work describes various aspects of the program, determines appropriate parameters adequate for both unique and characteristic GSR particle identification, and evaluates the reliability of data obtained. Samples are collected via the tape lift method from test-firings of .38, .32, .25, and .22 caliber handguns at time after firing intervals of 0 to 6 h. Unique GSR particles are consistently and correctly identified by this method on tape lift samples taken up to 4 h after firing. False positive results of unique GSR particles are not encountered on control handblank samples. This technique appears to provide the forensic science community with an operator-free method of reliable GSR particle search and an improved analyst-time-per-case ratio.

Electron Probe Microanalysis↗
PubMed1997

The effect of in vitro modeling conditions on the surface reactions of bioactive glass.

Using one parametric variation in solution composition, this paper documents that the surface reactions on bioactive glass (BG) 45S5 are exquisitely dependent upon the modeling conditions. The solutions used were 0.05 M tris hydroxymethyl aminomethane/HCl (tris buffer), tris buffer complemented with plasma electrolyte and/or serum, and serum. The reacted surfaces were analyzed using Fourier transform infrared (FTIR), scanning electron microscopy (SEM) with energy dispersive X-ray analysis (EDXA), and Rutherford backscattering spectroscopy (RBS). Post-immersion solutions were analyzed for changes in Ca and PO4 concentrations. After a short immersion (3 h), a crystalline, carbonated hydroxyapatite (c-HA) layer formed only in tris. Reaction surfaces of different structure, morphology, and composition were observed after various short and longer term immersions in all other solutions. They comprised two layers with the layer in contact with the bulk consisting mainly of Si; the outer layer, composed of Si, Ca, and P, was amorphous, and had a Ca/P ratio of about 1. Serum proteins adsorbed on the BG surfaces at the early stages of the solution-mediated BG reactions. Formation of a crystalline c-HA layer was delayed up to three or more days in solution with plasma ions. In the presence of serum, only amorphous surfaces composed of Si, Ca, and P were observed for any time up to seven days of immersion. The present data suggest that serum proteins adsorb in tandem with the occurrence of solution-mediated reactions leading to formation of a silica-gel. Amorphous Ca-P phases accumulate in the Si-rich matrix. Furthermore, the present data, in conjunction with the data published before, suggest that physicochemical and cell-mediated reactions occur in parallel to form the glass-tissue interfacial layer.

Biocompatible Materials↗
PubMed2005

A microprobe analysis of inorganic elements in Halobacterium salinarum.

Halobacterium salinarum were grown on peptone agar containing 4.28 M NaCl, 0.036 M K and other salts. Stationary phase organisms were lifted onto carbon planchets, freeze-dried, carbon coated and examined in a scanning electron microscope equipped with an X-ray spectrometer. Intracellular element concentrations (mol/kg H(2)O) were determined using a bulk analysis program with appropriate standards. The cell K concentration was 110 times that of the medium. For Na this value was 0.3 and for Cl, 1.1. When Rb was present in the medium, its intracellular concentration was 77 times higher than the external value. The cation minus anion value suggests a high fixed negative charge, 0.72 equivalents. Intracellular apparent dielectric constants were calculated using cellular EMFs derived from the literature, and sodium concentration. The determined values ranged from 22-28 (vs 80 for normal water) suggesting phases of structured cell water. Ionic distributions in these extremophiles are treated according to the classical principles elucidated by Willard Gibbs and represents a heterogeneous system in thermodynamic equilibrium with the hypersaline environment. Factors to be considered are: (1) composition of Halobacterium and its immobile negative charge; (2) the physicochemical properties of the individual ions (charge, ionic radius, hydration energy, standard chemical potential); (3) the dielectric constant of the dispersion medium (water); and (4) the binding of ions, particularly potassium.

Chlorides↗
PubMed1997

Apatite layer-coated titanium for use as bone bonding implants.

For development of thin bioactive coatings on metal implants, a dense and uniform apatite layer was coated onto titanium (Ti) implants in situ by using a new biomimetic method, which is composed of apatite nucleation and growth steps in simulated body fluid (SBF). Analysis of the coatings by thin film X-ray diffraction and scanning electron microscopy-energy dispersive X-ray microanalysis (SEM-EMPA) before implantation showed that its characteristics were very similar to those of natural bone. The coated and uncoated rectangular plates were bilaterally implanted into the tibial proximal metaphyses of rabbits. After 6, 10 and 25 weeks post-implantation, the bone bonding and bone formation at the bone-implant interfaces were evaluated by a detachment test and undecalcified histological examination. Mechanical testing in tension showed that the failure load of apatite layer-coated Ti implants was significantly higher than that of uncoated control at each time period (all P < 0.001). Histologically, it was shown that bone was deposited directly onto the apatite coating without any intervening soft tissue, while in the paired controls, interpositional soft tissue was seen at the bone-implant interface. By SEM-EPMA, a uniform calcium- and phosphorus-rich layer was detected between the coated implants and bone, but not in uncoated controls at either earlier or later time periods. The results indicate that the apatite layer deposited on Ti in situ may significantly increase the bone bonding strength by providing a bioactive surface, which allows for an early bone apposition to the implant. In addition, the apatite layer-coated Ti produced by the biomimetic process may fulfil the requirements of favourable thin coatings and strong adhesion at the metal-coating interface.

Analysis of Variance↗
PubMed1993

Anodic polarization behavior and microstructure of a gallium-based alloy.

A gallium-based alloy (GA) that was developed as a substitute for dental amalgam was investigated for anodic polarization behavior in deoxygenated Ringers solution, 37 degrees C. The related microstructures were examined and microanalyses were conducted. Four polarization tests were conducted by scanning from -300mV to +1,000 mV (vs. SCE) at 2 mV/s. Polarization of the first sample (GA-1) was stopped after the first anodic dissolution peak (-100 mV, 1.5-2.0 x 10(-3) A/cm2). The fourth sample (GA-4) was interrupted at the secondary peak (+1000 mV, 0.3 A/cm2). It was found that (1) the early stage of the first peak is related to selective dissolution of divalent tin ions, followed by a dissolution of Ga. Transmission electron diffraction (TED) identified the brown corrosion product as Ga2O3; (2) the GA-4 sample was covered with the white corrosion product of mainly Sn+4, identified as SnO2. In addition, the current density of the GA sample when coupled with a high-copper dental amalgam was 0.03 A/cm2 (with +1,000 mV) at the second peak which was about a ten times lower value than for the uncoupled sample; (3) the uncoupled gallium alloy and gallium alloy coupled with a high-copper dental amalgam showed 10(3)-10(4) times higher anodic current density than that of an uncoupled high-copper dental amalgam, suggesting that the gallium alloy is more corrosion prone.

Corrosion↗
PubMed1986

Inorganic particulates associated with pulmonary alveolar proteinosis: SEM and X-ray microanalysis results.

Twenty-four cases of pulmonary alveolar proteinosis (PAP) were studied by light microscopy (LM) and scanning electron microscopy (SEM) to test the hypothesis that PAP was related to silica exposure. Increased numbers of birefringent particles (vs. controls) were found in 78% of PAP cases. SEM was used to locate inorganic particulates in situ, which were individually analyzed using energy dispersive X-ray analysis. When analyzed as an aggregate group of cases, no specific inorganic particulate was evidently associated with the PAP reaction. However, analysis of individual cases revealed more specific associations. The concentration of particles determined by SEM exceeded that found by LM by a factor ranging from 2.7 to 964. The concentration of inorganic particulates per cm3 in the areas of PAP ranged from 1.3 X 10(7) to 1.02 X 10(9). Controls all had less than 10(7) particles per cm3. Available environmental history correlated well with particulate analysis results, e.g., silica in a sandblaster, metal fumes in a welder, and cement particles in a cement finisher. Particulates with unique composition were also found in cases with unavailable histories, e.g. metal fumes suggestive of welding or soldering exposure, silicates suggestive of fine particle exposure (greater than 50% of particles less than 1 micron). Only 1 case (the sandblaster) showed greater than 50% of the particles to be silica. Of the 5 infants with PAP, 3 showed the major particulate to be talc, and 1 had evidence of toxic cadmium selenide fume exposure. These results are consistent with the hypothesis that PAP, at least in the majority of cases, is associated with exposure to small inorganic particulates of several types.

Electron Probe Microanalysis↗
PubMed1980

A rapid, routine technique for the X-ray microanalysis of microincinerated cryosections: an SEM study of inorganic deposits in tissues of the marine gastropod Littorina littorea (L.).

A procedure is described that prepares chemically untreated biological sections for X-ray microanalysis in the scanning electron microscope (SEM). The method aims to retain and localize labile components in tissue sections by a procedure that is both rapid and routine. Large quantities of fresh tissue can be processed for analysis within a single day. Thick cryosections are cut with a steel knife in a conventional cryostat, freeze-dried, and then ashed by either low or high temperature incineration procedures. Controlled microincineration attenuates the organic matrix to reveal sufficient surface relief for effective SEM of some cytological structure and microanalysis of the residual inorganic components. The detectability of various elements is enhanced because the relative concentrations in the residues are increased and the level of nonspecific background in the X-ray spectra is reduced. The technique is applied to different tissues from the visceral complex of the marine prosobranch Littorina littorea. In animals exposed to elevated levels of zinc it can be demonstrated tht the metal is localized both as an insoluble form in granules and as a labile form within the cytoplasm. Other metals, including magnesium, potassium, calcium, manganese, and iron, have been identified and localized. The effectiveness of this technique for retaining labile elements is compared, in outline, with that of conventional fixation procedures.

Animals↗
PubMed1995

Effects of citric acid treatment on the Ca, P and Mg contents of human dental roots.

An electron microprobe analyser associated with a scanning electronic microscope was used to measure Ca, P and Mg contents of the human dental root hard tissues, before and after a citric acid treatment (pH = 1). The measurements were made on transverse sections through the cervical 1/3 of the molar roots. The measurements were performed at the following 8 levels: the internal cementum, the cementum-dentine junction, cementum-related dentine, 4 external dentine levels located at 220 microns, 420 microns, 620 microns and 820 microns from the cementum-dentine junction, and finally the juxta-pulpal dentine. After the citric acid treatment, the losses in Ca and P, but not in Mg, varied significantly with the level; an acid-resistant dentine layer of approximately 600 microns was found under the cementum-dentine junction. An increase in the Ca/P ratio was also observed in this layer. Since this external dentine zone is less demineralized by the citric acid than the underlying dentine, the collagen matrix in this region may also be less exposed. These differences in the response to citric acid that depend on the distance from the root surface may explain the unpredictability of periodontal healing after citric acid treatment of diseased root surfaces, because the amount of tissue removed during root planing is not constant.

Calcium↗
PubMed1984

Microdissection--elemental analysis of the mineralizing growth cartilage of the normal and rachitic chick.

The concentrations of elements in avian growth cartilage were studied by electron probe x-ray emission microanalysis (EDX). The cartilage was prepared for analysis by freezing, freeze-fracturing, freeze-drying, and carbon coating techniques. Cells and matrix fragments were removed from the tissue by microdissection with a tungsten needle in a scanning electron microscope (SEM) equipped with a real-time stereoscopic viewing facility. The samples were analyzed in the same SEM by EDX. Elemental analyses were performed on each fragment at a distance from the tissue sample, and hence background radiation due to the sample was eliminated. An important finding was that the intracellular potassium concentration of chondrocytes in calcified cartilage was similar to the levels in the premineralized zones. This observation supports the view that chondrocytes do not die in the process of, or as a consequence of, mineralization of the surrounding matrix. Calcium peaks were seen in the matrix at all levels and in chondrocytes immediately prior to mineralization. In contrast, phosphorus levels were always high in cells and low or absent from the premineralized matrix. At the mineralization front the appearance of a phosphorus peak in the matrix just preceded the deposition of mineral. We propose that the transfer of phosphorus from cell to matrix is a rate-limiting step in mineralization. Finally, when rachitic and normal cartilage were compared, little difference was seen in the profile of either intracellular or extracellular elements. However, in rickets the mineralized matrix remained soft in consistency. We suggest that this may reflect a phosphorus-related calcification defect that prevents growth and interlocking of the apatite crystallites.

Animals↗
PubMed1995

Scanning transmission electron microscopy/energy-dispersive spectroscopy analysis of the dentin adhesive interface using a labeled 2-hydroxyethylmethacrylate analogue.

In an attempt to compare the morphology of the dentin adhesive interface and the wetting and penetration of the adhesive in relation to the dentin surface, we studied four dentin adhesive systems using scanning transmission electron microscopy (STEM) and energy-dispersive spectroscopy (EDS). 2-Hydroxyethylmethacrylate (HEMA), a monomer common to many commercial dentin adhesive systems, was altered to produce a thiolated analogue (HETMA). Sulfur, traceable by EDS and STEM, was substituted for the oxygen atom in the backbone of the HEMA molecule. The resulting analogue, with solubility parameters and other wetting and physical properties very similar to those of HEMA, was applied to four sets of tooth specimens, each pre-treated with a different primer or etchant. Three separate pre-treatments--nitric acid, maleic acid, and citric acid/ferric chloride--created a demineralized zone approximately 1 to 3 microns thick at the dentin surface. The HETMA was found to permeate freely into this zone when either of the latter two pre-treatments was used. However, the band of dentin that was demineralized by the nitric acid pre-treatment appeared impermeable to the HETMA. The fourth pre-treatment, an alcohol-based solution including the phosphorus acid ester PENTA and HEMA, modified the smear layer of the tooth slightly and did not appear to demineralize the dentin. HETMA applied to the specimens pre-treated with PENTA and HEMA was clearly in intimate contact with the dentin or modified smear layer; however, it did not penetrate or diffuse into these areas. It did flow into the dentinal tubules, as was also evident with each of the other systems. It was concluded that the acid pre-treatment of the dentin greatly influenced the wetting behavior of the dentin adhesive and thus could substantially affect the resultant bond strength of the dentin adhesive systems.

Acid Etching, Dental↗