S.E.P. Dowker

Rietveld refinement of the crystallographic structure of human dental enamel apatites

Abstract Rietveld refinements using 12 sets of X-ray diffraction powder data from milligram samples of human dental enamel provide detailed information about the structure and composition of enamel apatite. The principal difference in atomic parameters between enamel apatite and Holly Springs hydroxylapatite is in O2, which is reflected in a reduction in the P-O2 bond length of 0.085 Å and PO4 volume by 3.6%. Modeling the hexad axis scattering with a single OH⁻ ion gives a 0.089 Å shift of the ion further away from the mirror plane at z = 1⁄4. The known distributed electron density along the hexad axis in enamel has been confirmed by direct comparison with synthetic hydroxylapatite. Although the CO32- ion position could not be determined directly, evidence for partial replacement of PO43- by CO32- ions came from an 8% diminution of the P site occupancy compared with that in stoichiometric hydroxylapatite. The observed reduction in the P-O2 bond length and PO4 volume in enamel is also consistent with this substitution. The loss of negative charge caused by CO32- replacing PO43- ions and loss of OH⁻ ions is compensated by loss of Ca2+ ions from Ca2 sites. The calculated density from the X-ray results is 3.021 g/cm3, in agreement with deductions from previous chemical analyses.

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.

Preparation and characterisation of monoclinic hydroxyapatite and its precipitated carbonate apatite intermediate

Five 100 g batches of a carbonate apatite (the intermediate) were produced by heating an aqueous slurry of CaCO3 and CaHPO4 with an overall Ca/P mole ratio of 5/3 with vigorous stirring. Each intermediate produced by boiling off water was heated in vacuum at 1100 degrees C to remove carbonate, then steamed at 900 degrees C to ensure complete hydroxylation. Comparison of calculated and observed X-ray diffraction patterns showed final products containing 50-100 wt% monoclinic hydroxyapatite (remainder hexagonal). Rietveld refinements in P6(3)/m gave structures similar to several hydroxyapatite standards, including NIST SRM 2910, although there was no evidence from X-ray diffraction that the latter was in the monoclinic form. Refinements from standards and final products were slightly different from published single crystal data for Holly Springs hydroxyapatite. This is attributed to known impurities in mineral hydroxyapatite and indicates that parameters from the Rietveld refinements are closer to the true values for pure hydroxyapatite. Rietveld refinements for intermediates showed small, but significant differences from the final product, the largest being in O1x, O2x and O(H)z. All P-O bond lengths were shorter than in the final product, resulting in a 3.2% lower PO4 tetrahedron volume. The occupancies of P and Ca(2) were reduced. These differences are attributed to partial replacement of PO4(3) by CO3(2-) ions.

Synchrotron X-Ray Microtomographic Investigation of Mineral Concentrations at Micrometre Scale in Sound and Carious Enamel

Synchrotron X-ray microtomography (XMT) was used to measure the linear attenuation coefficient (LAC) for 1.9-µm sidelength voxels within approximal brown spot lesions and sound human enamel. XMT demonstrated three-dimensional features, notably sheets with ∼30 µm periodicity having low LAC, identified as regions of demineralization corresponding to Retzius lines. Quantitative three-dimensional measurements of mineral concentration, derived from LAC with assumption of a single model composition, were consistent with previous measurements of sound and carious enamel from microradiographic projections. The uncertainty in measurements of mineral concentration and mineral fraction volume was investigated by modelling enamel with a range of composition and component densities. This analysis showed that, although mineral concentration can be determined from LAC with an error of <0.2 g cm–3, the variation in pore fraction volume within caries lesions cannot be reliably determined from X-ray attenuation measurements alone.

Longitudinal Study of the Three-Dimensional Development of Subsurface Enamel Lesions during in vitro Demineralisation

A longitudinal study was made of the 3D development of subsurface enamel lesions in whole human molars. X-ray microtomography (XMT) was used to measure the 3D distribution of linear attenuation coefficients in the tissue at 8–15 stages during cumulative times of 36–107 days demineralisation through ∼1-mm-wide windows. Although lesion morphology was consistent with preferential anisotropic dissolution parallel to enamel prisms at the advancing front, detail (at a scale of ∼100 µm) varied in relation to exposed surface sites separated by <1 mm. The distribution of mineral in the most superficial region varied across the exposed face of each lesion. Within lesions, localised foci of low mineral concentration (at a scale of ∼200 µm) retained their general form through successive stages of demineralisation before coalescing. The most advanced regions within a lesion seemed to correspond with surface regions with lowest mineral concentration. These findings indicate that local variations in fractional pore volume of partially demineralised enamel influence the subsequent spatial development of lesions.

Effects of sodium hypochlorite solution on root dentine composition

Sodium hypochlorite (NaOCl) solution,≤5% w/v available chlorine (abbreviated subsequently to %), is widely used as an irrigant in root canal treatment of teeth, so its effects on dentine are of clinical importance. The effects of ∼0.5%, 3% and 5% NaOCl solution on the composition of root dentine were studied at ambient temperature. For dentine powder treated for 30 min, depletion of the organic phase was confirmed by infrared spectroscopy. Apatite lattice parameters showed no significant change, but NaCl was also detected by X-ray powder diffraction. The low solubility of apatite mineral in the NaOCl solutions was demonstrated by the constant weight of bulk enamel specimens immersed for seven days. The stability of the mineral phase was confirmed by scanning microradiography (SMR), an X-ray attenuation method employing photon counting. Repeated SMR measurements of the local mineral content of bulk samples of root dentine and a synthetic hydroxyapatite aggregate during exposure to pumped NaOCl solutions for 100 h showed no mineral loss. As predicted from apatite chemistry, reaction of NaOCl with the mineral phase can be excluded as a primary factor in changes in mechanical properties of treated dentine. Effects of retention of NaCl on endodontic sealants requires further investigation.

DETERMINATION OF MINERAL CONCENTRATION IN DENTAL ENAMEL FROM X-RAY ATTENUATION MEASUREMENTS

The mineral content of dental enamel is commonly measured by X-ray attenuation experiments. Most studies have used contact microradiography in which intensities are measured with photographic film which is convenient and gives high spatial resolution. However photon counting intensity measurements are to be preferred in many experiments (longitudinal and scanning microradiography, and microtomography), as illustrated here, because they have a larger dynamic range and greater sensitivity to small intensity changes. Additionally, the detector and specimen are well separated which allows the pseudo-continuous study of de- and remineralization. The mineral content is often quoted as 95 wt% or 87 vol% hydroxyapatite for permanent human enamel. This determination from attenuation experiments requires accurate values of elemental mass attenuation coefficients and a number of assumptions. The effects of possible choices of these are considered and it is shown that the most important is the density of enamel mineral used in conversion of wt% to vol%. If the density is taken as 2.99 g cm(-3), as recently suggested (J.C. Elliott, Dental Enamel, Ciba Foundation Symposium 205, Wiley, Chichester, pp. 54-72, 1997), instead of 3.15 g cm(-3) as for hydroxyapatite, the calculated vol% is approximately 93 instead of approximately 87.

Application of scanning microradiography and x-ray microtomography to studies of bones and teeth.

In scanning microradiography (SMR), a thin section is stepped across a 15-μm diameter X-ray beam and the transmitted intensity measured at each point. This technique has permitted more accurate measurements of the spatial variation of the mineral concentration in sections of dentin and enamel than conventional photographic microradiography. Moreover, because the section is not in close contact with an emulsion, SMR allows continuous study while the specimen is bathed in a reaction solution. The present studies have been particularly directed to gaining an understanding of the formation and repair of carious lesions in teeth: one particular puzzle is subsurface demineralization, in which the initial loss of mineral appears to take place some 20 to 50 μm below the tooth surface. SMR studies are reported here on the demineralization in dilute acids and the subsequent partial remineralization in supersaturated calcium phosphate solutions in model systems for dental caries. In order to develop a theoretical model for de- and remineralization of carious lesions, it is necessary to quantify transport processes within the tooth. To this end, we are developing a method of measuring effective diffusion coefficients of strongly X-ray-absorbing ions in water within permeable solids in which the diffusion coefficient varies with position. The method uses sequential concentration/distance profiles determined by SMR. As a test, diffusion coefficients of potassium iodide in water within a permeable glass frit have been measured. X-ray microtomography (XMT) can be carried out by adding an axis of rotation to the SMR apparatus. Using this method, linear absorption coefficients, and hence mineral concentrations, can be measured in 15 X 15 X 15-μm3 voxels. This has advantages over SMR in that superposition within the depth of the section and errors in determining its thickness are avoided. XMT studies of de- and remineralization similar to those described above for SMR, and also XMT studies of the variation in mineral concentration in the cortical bone of a rat femur along its length, are reported.

Optical Profilometric Study of Changes in Surface Roughness of Enamel during in vitro Demineralization

The application of non–contact optical profilometry for non–destructive study of changes in the surface roughness of natural enamel surfaces during in vitro demineralization was investigated. Repeated measurement of profiles of the same enamel surfaces after successive demineralization episodes was achieved by a kinematic specimen mount that could be removed, then accurately repositioned. Surface roughness parameters and reflectivity of natural enamel surfaces exposed to either a demineralizing solution (test) or deionized water (control) for up to 6 days were measured at 6– to 24–hour intervals. The results prior to demineralization showed that surface roughness varies with position on the enamel surface. During demineralization there was an approximately linear increase in surface roughness for the first 70 h followed by somewhat erratic behaviour, and a saturating exponential with time increase in reflectivity for the first 70 h which subsequently remained constant. Changes in enamel surface during in vitro demineralization were also observed using scanning electron microscopy.

CRYSTAL CHEMISTRY AND DISSOLUTION OF CALCIUM PHOSPHATE IN DENTAL ENAMEL

Abstract The mineral component (at least 95 wt. %) of dental enamel is hydroxyapatite (hydroxylapatite) with multiple substitutions. The biogenic origin of enamel is reflected in the unusual ribbon-like morphology of the crystals, which are extremely elongated in the c-axis direction, and their organized arrangement within the tissue. The study of enamel dissolution has been driven by the very high prevalence of dental caries. In enamel caries, the initial demineralization results in subsurface dissolution of mineral. While the surface remains intact, reversal of the lesion by remineralization is possible. Problems of understanding the physico-chemical processes in enamel demineralization include the general problems concerning the structure and chemistry of apatites formed in aqueous media. Added to these are the general problem of dissolution in an inhomogeneous porous medium and the complication that enamel apatite has a naturally variable composition which changes during demineralization. The use of model systems in caries research is illustrated by reference to X-ray absorption studies of enamel and synthetic analogues.