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Biomedical subjects

J C Elliott

Publications and source records attributed to J C Elliott.

At least 19 recordsLinked to original sources

Cells for the study of acidic dissolution in packed apatite powders as model systems for dental caries.

A number of different designs of cells have been developed for the study of dissolution processes in packed apatite powders. Basic design requirements were that no deleterious processes, such as high temperature sintering or binding agents, were involved, and that there was maximum opportunity for experimental study. One design used alternating filter paper discs and apatite layers (typically 42 mg). At the end of an experiment, the cell could be disassembled and infrared spectra and X-ray powder diffraction patterns made of individual layers. Cells without filter paper discs were also made, but terminal sampling at well-defined depths was more difficult or impossible. The cells were constructed from poly(methylmethacrylate) so the course of dissolution could be monitored by radiography. Subsurface loss of apatite was almost always seen after from 2 weeks to 8 months exposure to buffer solutions at pH 3.0-5.5. The greatest loss of apatite was typically 0.4-2 mm below the surface, which is at a larger distance than usually seen for dental enamel. This may be attributable to the low packing density (typically 35-60 vol%) found in the present systems compared with enamel.

Acids

Theoretical explanation of the relationship between backscattered electron and x-ray linear attenuation coefficients in calcified tissues.

X-ray absorption and backscattered electron (BSE) microscopies are two commonly used techniques for estimating mineral contents in calcified tissues. The resolution in BSE images is usually higher than in x-ray images, but due to the previous lack of good standards to quantify the grey levels in BSE images of bones and teeth, x-ray microtomography (XMT) images of the same specimens have been used for calibration. However, the physics of these two techniques is different: for a specimen with a given composition, the x-ray linear attenuation coefficient is proportional to density, but there is no such relation with the BSE coefficient. To understand the reason that this calibration appears to be valid, the behaviour of simulated bone samples was investigated. In this, the bone samples were modelled as having three phases: hydroxyapatite (Ca10(PO4)6(OH)2), protein, and void (either empty or completely filled with polymethylmethacrylate (PMMA), a resin which is usually used for embedding bones and teeth in microscopic studies). The x-ray linear attenuation coefficients (calculated using published data) and the BSE coefficients (calculated using Monte Carlo simulation) were compared for samples of various phase proportions. It was found that the BSE coefficient correlated only with the x-ray attenuation coefficient for samples with PMMA infiltration. This was attributed to the properties of PMMA (density and mean atomic number) being very similar to those of the protein; therefore, the sample behaves like a two-phase system which allows the establishment of a monotonic relation between density and BSE coefficient. With the newly developed standards (brominated and iodinated dimethacrylate esters) for BSE microscopy of bone, grey levels can be converted to absolute BSE coefficients by linear interpolation, from which equivalent densities can be determined.

Bone Density

X-ray microtomography: nondestructive three-dimensional imaging for in vitro endodontic studies.

Article shows the application of a laboratory x-ray microtomography system, a miniaturized form of conventional computerized axial tomography, to the study of root canal morphologic characteristics and changes in the course of root canal treatment in extracted teeth. After reconstruction of the three-dimensional images, the IDL software package (Research Systems, Inc., Colorado) was used to obtain cross-sectional slices of the tooth and three-dimensional views of rendered surfaces of constant mineral density. The root canal systems and changes in these were imaged at a resolution (cubic voxel side-length) of approximately 40 microns.

Anatomy, Cross-Sectional

X-ray microtomography: 3-dimensional imaging of teeth for computer-assisted learning.

X-ray microtomography (XMT), a miniaturised form of computed tomography, has been used to generate 3-dimensional volume data sets of the X-ray absorption of human teeth in vitro, with a resolution of approximately 40 microns (cubic voxel sidelength). Examples are presented of images relevant to dental morphology, cariology, and cavity preparation and restoration. Applications of XMT imaging to dental education are discussed in the context of new approaches to visual learning through computer-assisted methods.

Computer-Assisted Instruction

Structure, crystal chemistry and density of enamel apatites.

The apatitic calcium phosphate crystals in dental enamel are too small for single crystal diffraction studies so the only possible direct structure determination must use whole-pattern-fitting Rietveld analysis of X-ray and neutron powder diffraction patterns. As a result, aspects of the structure are not known in detail. Further structural information can be obtained by consideration of published chemical analyses and infrared studies, taking into account studies of the crystal chemistry of synthetic apatitic analogues of enamel apatite. The apatitic constitutional water and total water content of enamel are particularly important, but there are difficulties in their determination. Making reasonable assumptions, a number of models of the unit cell can be derived. The weight per cent (including constitutional water) and density of the enamel apatite crystals for the most probable model are about 98 wt.% and 3.0 g cm-3, respectively. The apatite volume per cent calculated from these values is about 96%. The weight per cent and volume per cent of enamel apatite are higher than normally accepted values because of inclusion of constitutional water and use of a density for enamel apatite that takes into account its known lattice expansion over hydroxyapatite and probable lattice vacancies.

Animals

A comparison of the mineral content of enamel and dentine in human premolars and enamel pearls measured by X-ray microtomography.

Mineral content gradients in two composite enamel pearls from permanent human upper molars were measured by X-ray microtomography (XMT) at a resolution of 15-30 microns. This non-destructive microscopic technique was used to make 15-microns thick XMT slices with 100-microns separation through one pearl and 250-microns separation through the other. Average mineral contents were calculated from the linear absorption coefficients determined from regions of the XMT slices assuming the inorganic component to be calcium hydroxyapatite. These values were compared with similar XMT studies of coronal enamel and dentine of upper permanent premolars. A mineral content gradient in the pearls, reducing from the enamel surface to the amelodentinal junction, was found; this was similar to that observed in the coronal enamel of the upper premolar. The mineral contents in the surface and deeper enamel regions of the pearl were similar to those observed in premolar enamel. In contrast, the mineral content for the dentine of the pearl was greatest at the amelodentinal junction, i.e. the gradient was in the opposite direction to that observed in premolar dentine. These results suggest that the process of mineralization of the pearl dentine differs from that in permanent control dentine. In addition, gradients in enamel and dentine mineral contents reducing from the tip of the pearl to the base of the pearl were found.

Bicuspid

Characterization of human lymphoid cell lines GM9947 and GM9948 as intra- and interlaboratory reference standards for DNA typing.

The incorporation of reference DNA is crucial to the validation of any DNA typing protocol. Currently, reference DNA standards are restricted to molecular size DNA ladders and/or tumor cell line DNA. Either of these, however, presents some limitations. We have rigorously characterized two Epstein-Barr virus (EBV)-immortalized human lymphoid cell lines--GM9947 (female) and GM9948 (male)--to determine their suitability as alternative in-line standards for three widely employed allele profiling strategies. Twenty-one highly polymorphic VNTR-based allelic systems (7 RFLPs, 2 AmpFLPs, and 12 STRs) distributed over 12 chromosomes were scrutinized along with 3 gender-based discriminatory systems. The genetic stability of each locus was confirmed over a period of 225 in vitro population doublings. Allele size estimates and degree of informativeness for each of the 21 VNTR systems were compiled. The reproducibility of allele scoring by traditional RFLP analyses, using both cell lines as reference standards, was also verified by an interlaboratory validation study involving 13 analysts from two geographically distinct forensic laboratories. Taken together, our data indicate that GM9947 and GM9948 genomic DNAs could be adopted as reliable reference standards for DNA typing.

Base Sequence

Mineral concentration gradients in rat femoral diaphyses measured by X-ray microtomography.

The bone mineral concentrations of five rat femora were measured as a function of distance from the distal metaphysis by quantitative X-ray microtomography (XMT) at a resolution of approximately 23 x 23 x 15 microns3. Assuming the mineral phase of bone to be hydroxyapatite, Ca10 (PO4)6(OH)2, the mean cortical mineral concentration (CM) per transverse section was found to range from 1.33 to 1.47 g cm-3. Detectable variations in the bone mineral concentration between sections of femora from different animals could not be attributed to the age when the particular animal was sacrificed. An increase in CM with distance, L, from the distal growth plate was observed and a saturating exponential equation, CM = a - be-alpha L, was used to describe the changes. Each section of bone tissue was considered as a population of elementary volumes of bone (EVB) and L was related to the age of the EVB (TEVB). A simple model for the mineralization process of an EVB was then proposed. Each newly formed EVB accumulated mineral rapidly to give an initial mineral concentration of approximately 1.3 g cm-3 (parameter a-b). Their mineral concentrations then increased asymptotically to approximately 1.5 g cm-3 (parameter a) with a time constant of approximately 330 days. This slow maturation process is attributed to Ostwald ripening of the bone crystals with further crystal growth using ions from the extracellular fluid.

Analysis of Variance

Deciduous enamel defects in low-birth-weight children: correlated X-ray microtomographic and backscattered electron imaging study of hypoplasia and hypomineralization.

Enamel does not remodel, and disturbances occurring during development may remain in the tooth as a permanent record of the upset. Mineralization in prenatal and postnatal deciduous enamel was studied in the shed deciduous incisors of low-birth-weight (LBW: < 2kg) children. The specific objective was to gain further insight into the mechanism of formation of developmental defects of enamel. Sections at a resolution of 22-40 microns were reconstructed using X-ray microtomography (microCT) giving absolute measurements of linear absorption coefficient for AgK alpha radiation. Detail to ca. 1 micron resolution was obtained using automated, digital backscattered electron (BSE) imaging of PMMA-embedded material. Matching the histograms of BSE and microCT images made possible the calibration of the mean atomic number-dependent signal in the BSE images. The comparison of abnormal, affected enamel regions and post-recovery, normal, unaffected regions could be made in the same teeth, since these zones were easily recognized from the distribution of hypoplasia and hypomineralization. The microCT values, converted to calculated mineral densities, ranged from 2.3 g cm-3 to 2.6 g cm-3 in LBW hypoplastic, and between 2.65 and 2.78 g cm-3 in control primary enamel and post-defect, post-natal LBW enamel. Hypoplasia with or without minimal hypomineralization indicated recovery of the ameloblasts in the maturation phase. Disturbance during late matrix formation and early maturation resulted in hypoplasia and hypomineralization.

Child, Preschool

Liquid scintillation sample analysis in microcentrifuge tubes.

Local regulations prohibiting drain disposal of "biodegradable" liquid scintillation cocktails prompted investigation of volume reduction for these materials. Microcentrifuge tubes were used with aqueous and filter media samples of 3H, 14C, 32P, and 125I. Backgrounds, counting efficiencies, figures of merit, and spectral distributions obtained for microcentrifuge tubes compared favorably to conventional vials. Differences in 32P spectra for solid support samples appeared related to filter material and sample volume. Decreases in sample costs and waste volume and disposal costs were approximately 50-75%.

Radioactive Waste

Scanning microradiographic study of the kinetics of subsurface demineralization in tooth sections under constant-composition and small constant-volume conditions.

The kinetics of subsurface demineralization of tooth sections has been studied in real-time by scanning microradiography (SMR). Demineralization was carried out: (1) with a large volume of solution buffered to pH = 4 to maintain a constant composition; and (2) in a small constant volume (approximately 3 mL), buffered initially at pH = 4, so that the degree of saturation at the tooth surface increased as the tooth dissolved. At constant composition, the change in lesion depth, Y, with time, T, followed a linear relation, Y = a + bT, for T > 44 +/- 5 h. Before this time, the relation could be approximated by a linear one with different a and b constants. At constant volume, Y = q(1-e-r(T + s)) for all T, where q, r and s are constants. Similar relations, with different constants, were found for the mineral loss per unit area of lesion exposed to acid. These results showed that the process of demineralization under the rather severe conditions used was essentially a surface-controlled process. The change of slope at approximately 44 h and the presence of the constant s in the exponential function were attributed to a change in kinetics after formation of the surface layer.

Dental Enamel

Acoustic microscope study of the elastic properties of fluorapatite and hydroxyapatite, tooth enamel and bone.

Measurements of Rayleigh velocity and attenuation were taken in single mineral crystals of hydroxyapatite and fluorapatite at angular intervals relative to their c axes, using an acoustic microscope. These results are compared with the values that were calculated using the elastic constants of apatite from Yoon and Newnham [(1969) Am. Miner. 54, 1193-1197.] and Katz and Ukraincik [(1971) J. Biomechanics 4, 221-227.]. The slowness curves of various wave modes are plotted and discussed in relation to cross-coupling effects that were found to cause instability in measurements of attenuation for the c axes direction. Velocity measurements were taken in specimens of tooth enamel and bone. Here comparisons are made with the values that were calculated by modelling the 'z' scan response of the microscope, using published data for the elastic and acoustic properties. Comparisons are also made with the measurements on single crystals, since apatite is a major component of enamel and bone.

Animals

Evaluation of smear layers on serial sections of human dentin by means of electrochemical impedance measurements.

The alternating current impedance characteristics of a uni-directional porous glass membrane and serial sections of dentin from unerupted human third permanent molars (with and without smear layers) were measured. The membrane was equilibrated at 20 +/- 1 degrees C in a range of concentrations of unbuffered KCl, and the tooth sections were equilibrated in 0.1 mmol/L unbuffered KCl. Each section's impedance was measured for a 50-mV (rms) alternating potential difference applied over 1 Hz to 65 kHz. By means of complex non-linear least-squares regression analysis, the dc resistance of each specimen was determined. Scanning electron microscopy and subsequent image analysis enabled the cross-sectional area of the pores in the membrane and dentin tubule surface-openings to be measured. The results showed that the measured cross-sectional area of pores in the membrane was in good agreement with that calculated from the measured dc resistance and solution resistivity over the range of electrolyte concentrations. In dentin, the presence of smear layers on the cut surfaces increased the measured resistance. The resistance of the smear layer varied, suggesting that its thickness changed across the surface of the dentin. This was superimposed on regional variations in resistance of smear-free dentin which were found to be related to alterations in the number of tubules present and their cross-sectional area. The resistivities of the dentin tubule contents and smear layer material were generally lower than that of the hydrating electrolyte. It is postulated that this could be due to ions from partial dissolution of the mineral in the dentin.

Dentin

Subsurface demineralization in dental enamel and other permeable solids during acid dissolution.

Subsurface demineralization of dental enamel during acid dissolution has been reported many times, but its cause remains obscure. At first, the phenomenon was thought to result from the physical structure of enamel. More recent studies have shown that subsurface demineralization occurs in other permeable solids, indicating that there must be more fundamental factors involved in this curious effect. In order for this phenomenon to be investigated, dissolution experiments were carried out by means of real-time scanning microradiography in various systems, including enamel, or aggregates of hydroxyapatite (calcium, strontium, or barium), or hydroxides (calcium or magnesium). These were chosen to discriminate between effects of structure and composition. It was found that it was not possible for the demineralization observed in these systems to be attributed to a common feature. From this, it is concluded that subsurface demineralization in enamel and other mineralized tissues should not be ascribed to a single cause.

Acetates

Scanning and contact microradiographic study of the effect of degree of saturation on the rate of enamel demineralization.

The effect of degree of saturation with respect to hydroxyapatite (DSHA) on enamel section demineralization was studied by contact and scanning microradiography (CMR and SMR). Two aspects were studied: (1) the effect of different values of DSHA at constant pH, and (2) the effect of constant DSHA with variable pH. In the pH range (2.5-4.5) and time scale (0-120 h) studied, the DSHA was more important in determining the rate of enamel lesion progression than was the pH.

Adult

Regional distribution of mineral and matrix in the femurs of rats flown on Cosmos 1887 biosatellite.

We combined biochemical measurements with novel techniques for image analysis in the rat femur to characterize the location and nature of the defect in mineralization known to occur in growing animals after spaceflight. Concentrations of mineral and osteocalcin were low in the distal half of the diaphysis and concentrations of collagen were low with evidence of increased synthesis in the proximal half of the diaphysis of the flight bones. X-ray microtomography provided semiquantitative data in computer-generated sections of whole wet bone that indicated a longitudinal gradient of decreasing mineralization toward the distal diaphysis, similar to the chemistry results. Analysis of embedded sections by backscattered electrons in a scanning electron microscope revealed distinct patterns of mineral distribution in the proximal, central, and distal regions of the diaphysis and also showed a net reduction in mineral levels toward the distal shaft. Increases in mineral density to higher fractions in controls were less in the flight bones at all three levels, with the most distal cross-sectional area most affected. The combined results from these novel techniques identified the areas of femoral diaphysis most vulnerable to the mineralization defect associated with spaceflight and/or the stress of landing.

Animals

Electrochemical impedance characterization of human and bovine enamel.

The alternating current impedance characteristics of human and bovine enamel slices were measured in vitro. An electrochemical cell containing two platinum gauze electrodes and 0.01 mol/L KCl buffered to pH 7.4 at 20 degrees C was used. The electrodes were attached to a potentiostat that was connected to a microcomputer-controlled frequency-response analyzer. Measurements were made at discrete frequencies between 1 Hz and 65 kHz by application of a sinusoidal potential of 50 mV (rms) across the assembled cell. The impedance was calculated from the input potential and the resulting measured flow of current. For quantitative evaluation of these measurements, an equivalent circuit was postulated. It contained five passive electronic components and accurately modeled the different specimens. Values for individual electrical components in the equivalent circuit were estimated by complex non-linear least-squares regression analysis. This study demonstrated that it is possible to measure and quantitatively distinguish between the impedance characteristics of: permanent human, deciduous human, and bovine enamel, as well as the enamel from a variety of sites from human teeth, some of which had been extracted prior to being fully erupted. Proposals are made as to which physical properties in the specimens are modeled by individual components in the equivalent circuit.

Animals