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Detoxification of a highly toxic lead-loaded industrial solid waste by stabilization using apatites.

Apatites are known for their properties to immobilize lead contained in aqueous solutions or contaminated soils. In this study, apatites were examined as stabilization additives for lead-loaded industrial solid toxic wastes. The specific waste was the residue, obtained after thermal treatment of sludges (incineration), which was derived from tetraethyl lead fuel storage tanks. It was found to contain around 30 wt.% lead and 33 wt.% iron. Standard leaching tests (according to DIN 38414 S-4) were applied for the determination of leachability of metals from the ash and, thus, of chemical toxicity; the proposed leaching tests examined both initial and stabilized products in order to evaluate the effectiveness of the applied additives. The results obtained demonstrate the fact that lead concentrations in leachates, after the application of the proposed leaching tests using apatites as additives and with a ratio of 50% solid waste-50 wt.% apatite, could be reduced to the range of 1mg/l.

Adsorption↗

Analysis of self-assembly and apatite binding properties of amelogenin proteins lacking the hydrophilic C-terminal.

Amelogenins, the major protein component of the mineralizing enamel extracellular matrix, are critical for normal enamel formation as documented in the linkage studies of a group of inherited disorders, with defective enamel formation, called Amelogenesis imperfecta. Recent cases of Amelogenesis imperfecta include mutations that resulted in truncated amelogenin protein lacking the hydrophilic C-terminal amino acids. Current advances in knowledge on amelogenin structure, nanospheres assembly and their effects on crystal growth have supported the hypothesis that amelogenin nanospheres provide the organized microstructure for the initiation and modulated growth of enamel apatite crystals. In order to evaluate the function of the conserved hydrophilic C-terminal telopeptide during enamel biomineralization, the present study was designed to analyze the self-assembly and apatite binding behavior of amelogenin proteins and their isoforms lacking the hydrophilic C-terminal. We applied dynamic light scattering to investigate the size distribution of amelogenin nanospheres formed by a series of native and recombinant proteins. In addition, the apatite binding properties of these amelogenins were examined using commercially available hydroxyapatite crystals. Amelogenins lacking the carboxy-terminal (native P161 and recombinant rM166) formed larger nanospheres than those formed by their full-length precursors: native P173 and recombinant rM179. These data suggest that after removal of the hydrophilic carboxy-terminal segment further association of the nanospheres takes place through hydrophobic interactions. The affinity of amelogenins lacking the carboxy-terminal regions to apatite crystals was significantly lower than their parent amelogenins. These structure-functional analyses suggest that the hydrophilic carboxy-terminal plays critical functional roles in mineralization of enamel and that the lack of this segment causes abnormal mineralization.

Amelogenin↗

Evidence of hydroxyl-ion deficiency in bone apatites: an inelastic neutron-scattering study.

The novelty of very large neutron-scattering intensity from the nuclear-spin incoherence in hydrogen has permitted the determination of atomic motion of hydrogen in synthetic hydroxyapatite and in deproteinated isolated apatite crystals of bovine and rat bone without the interference of vibrational modes from other structural units. From an inelastic neutron-scattering experiment, we found no sharp excitations characteristic of the vibrational mode and stretch vibrations of OH ions around 80 and 450 meV (645 and 3630 cm(-1)), respectively, in the isolated, deproteinated crystals of bone apatites; such salient features were clearly seen in micron- and nanometer-size crystals of pure hydroxyapatite powders. Thus, the data provide additional definitive evidence for the lack of OH(-) ions in the crystals of bone apatite. Weak features at 160-180 and 376 meV, which are clearly observed in the apatite crystals of rat bone and possibly in adult mature bovine bone, but to a much lesser degree, but not in the synthetic hydroxyapatite, are assigned to the deformation and stretch modes of OH ions belonging to HPO(4)-like species.

Animals↗

Crystal alignment of carbonated apatite in bone and calcified tendon: results from quantitative texture analysis.

Calcified tissue contains collagen associated with minute crystallites of carbonated apatite. In this study, methods of quantitative X-ray texture analysis were used to determine the orientation distribution and texture strength of apatite in a calcified turkey tendon and in trabecular and cortical regions of osteonal bovine ankle bone (metacarpus). To resolve local heterogeneity, a 2 or 10 microm synchrotron microfocus X-ray beam (lambda = 0.78 A) was employed. Both samples revealed a strong texture. In the case of turkey tendon, 12 times more c axes of hexagonal apatite were parallel to the fibril axis than perpendicular, and a axes had rotational freedom about the c axis. In bovine bone, the orientation density of the c axes was three times higher parallel to the surface of collagen fibrils than perpendicular to it, and there was no preferential alignment with respect to the long axis of the bone (fiber texture). Whereas half of the apatite crystallites were strongly oriented, the remaining half had a random orientation distribution. The synchrotron X-ray texture results were consistent with previous analyses of mineral orientation in calcified tissues by conventional X-ray and neutron diffraction and electron microscopy, but gave, for the first time, a quantitative description.

Animals↗

Precursor decomposition and nucleation kinetics during platelike apatite synthesis.

Self-organization of calcium and phosphorus precursors in solution containing acetic acid and ethylene glycol produces a nanosized lamellar acetate-phosphonate hybrid containing two acetate and one phosphonate components. The lamellar morphology of the hybrid precursor is responsible the formation of platelike apatite product after thermal treatment at or above 400 degrees C. However a preliminary preheating stage (300 degrees C, 24 h) is crucial in determining the morphology of the apatite. Activation energy measurements by nonisothermal thermogravimetric analysis show that decomposition of the hybrid precursor involves at least two steps. Among the three components, it appears that the calcium acetate bidentate chelate component is stable below or at 300 degrees C. However, the calcium phosphonate and calcium acetate monodentate components are decomposed at this temperature. Above 360 degrees C, nuclear magnetic resonance and infrared spectroscopic data reveal the decomposition of more stable calcium acetate bidentate chelate. It is evident that the bond rupture of the bidentate calcium acetate species in the precursor results in the start of crystalline apatite formation but the other components must be decomposed by heating prior to this critical step in order to produce platelike apatite.

Apatites↗

The formation of a hydroxyl bond and the effects thereof on bone-like apatite formation on a magnesia partially stabilized zirconia (MgO-PSZ) bioceramic following CO2 laser irradiation.

For the purpose of improving the bioactivity of a magnesia partially stabilized zirconia (MgO-PSZ) and to explore a new technique for inducing OH group and apatite formation, a CO(2) laser has been used to modified the surface properties. The bioactivity of the CO(2) laser modified MgO-PSZ has been investigated in stimulated human fluids (SBF) with ion concentrations almost equal to those in human blood plasma. Some hydroxyl groups were found on the MgO-PSZ following CO(2) laser treatment with selected power densities. The surface melting on the MgO-PSZ induced by CO(2) laser processing provides the Zr(4+) ion and OH(-) ion, in turn, the incorporation of the Zr(4+) ion and the OH(-) ion creates the Zr-OH group on the surface. After 14 days of SBF soaking, the apatites formed on the MgO-PSZ with relatively high amount of hydroxyl groups generated by the CO(2) laser treatment, while no apatite was observed on the untreated with few hydroxyl groups. It exhibits that the Zr-OH groups on the MgO-PSZ surface is the functional groups to facilitate the apatite formation. The increased surface roughness provides more active sites, meantime, increased surface energy benefits to the adsorption and reaction on the surface.

Adsorption↗

Mycorrhizal weathering of apatite as an important calcium source in base-poor forest ecosystems.

The depletion of calcium in forest ecosystems of the northeastern USA is thought to be a consequence of acidic deposition and to be at present restricting the recovery of forest and aquatic systems now that acidic deposition itself is declining. This depletion of calcium has been inferred from studies showing that sources of calcium in forest ecosystems namely, atmospheric deposition and mineral weathering of silicate rocks such as plagioclase, a calcium-sodium silicate do not match calcium outputs observed in forest streams. It is therefore thought that calcium is being lost from exchangeable and organically bound calcium in forest soils. Here we investigate the sources of calcium in the Hubbard Brook experimental forest, through analysis of calcium and strontium abundances and strontium isotope ratios within various soil, vegetation and hydrological pools. We show that the dissolution of apatite (calcium phosphate) represents a source of calcium that is comparable in size to known inputs from atmospheric sources and silicate weathering. Moreover, apatite-derived calcium was utilized largely by ectomycorrhizal tree species, suggesting that mycorrhizae may weather apatite and absorb the released ions directly, without the ions entering the exchangeable soil pool. Therefore, it seems that apatite weathering can compensate for some of the calcium lost from base-poor ecosystems, and should be considered when estimating soil acidification impacts and calcium cycling.

Apatites↗

Apatite-mediated actin dynamics in resorbing osteoclasts.

The actin cytoskeleton is essential for osteoclasts main function, bone resorption. Two different organizations of actin have been described in osteoclasts, the podosomes belt corresponding to numerous F-actin columns arranged at the cell periphery, and the sealing zone defined as a unique large band of actin. To compare the role of these two different actin organizations, we imaged osteoclasts on various substrata: glass, dentin, and apatite. Using primary osteoclasts expressing GFP-actin, we found that podosome belts and sealing zones, both very dynamic actin structures, were present in mature osteoclasts; podosome belts were observed only in spread osteoclasts adhering onto glass, whereas sealing zone were seen in apico-basal polarized osteoclasts adherent on mineralized matrix. Dynamic observations of several resorption cycles of osteoclasts seeded on apatite revealed that 1) podosomes do not fuse together to form the sealing zone; 2) osteoclasts alternate successive stationary polarized resorption phases with a sealing zone and migration, nonresorption phases without any specific actin structure; and 3) apatite itself promotes sealing zone formation though c-src and Rho signaling. Finally, our work suggests that apatite-mediated sealing zone formation is dependent on both c-src and Rho whereas apico-basal polarization requires only Rho.

Actins↗

The influence of unstable signals for electron spin resonance dosimetry with synthetic A-type carbonated apatite.

Synthetic A-type carbonated apatite prepared in controlled conditions was irradiated at room temperature with 60Co gamma rays. The ESR spectrum was associated to axial CO2- and orthorhombic CO3- species. Radicals used as dose markers in biological apatites are long-lived paramagnetic species. The stability of the post-irradiation signal of A-type apatite was investigated for almost 2 years. Measurements showed variations in the spectra attributed to unstable CO3- species, which can be eliminated by thermal treatments at 100 degrees C for 24 h. Results indicated the potential use of an A-type carbonated apatite as a dosemeter.

Apatites↗

Apatite plaque particles in inner medulla of kidneys of calcium oxalate stone formers: osteopontin localization.

BACKGROUND: We have previously shown that interstitial plaque particles appear first in the basement membranes of thin loops of Henle and then in the interstitial space. However, it is not known if the plaque in the basement membrane of thin loops of Henle is of the same or different form than the interstitial plaque. Thus our purpose here is to detail the structure of the interstitial and membrane-bound plaque and explore the relationship of plaque apatite to osteopontin, a well-known crystal-associated urine protein. METHODS: Deep papillary biopsy tissue was studied from all 15 calcium oxalate stone formers and four nonforming subjects that we previously reported on [Evan et al, J Clin Ivest, 2003]. Routine light and transmission electron microscopy (TEM) as well as light microscopy and TEM immunohistochemical localization of osteopontin antibody were performed on all 19 subjects. RESULTS: In the basement membrane, plaque particles are individual and appear laminated with alternating light regions of crystal and electron-dense organic layers. In the interstitium, individual particles are not abundant but are instead aggregated to form regions of attached particles and in some regions what appears to be a fusion or syncytium in which crystal islands float in an organic sea. By light microscopy immunohistochemistry, osteopontin was localized to cells of the loops of Henle and collecting ducts as well as on sites of plaque. By immunoelectron microscopy, osteopontin immunogold label was found mainly on the surfaces of apatite crystal phase, at the junction of the crystal/organic layers. A similar immunogold labeling pattern was seen in the particles forming the syncytial islands of interstitial plaque. CONCLUSION: If indeed we accept the hypothesis that apatite plaque may be an anchored site on which calcium oxalate stones form and grow, the present work makes clear that it is unlikely that the surface of plaque presented to the final urine will be apatite crystal per se. However, our findings clearly show osteopontin is one of the crystal-associated urine proteins involved in the formation of the organic layers of the plaque particles.

Apatites↗

Control of apatite crystal growth by the co-operative effect of a recombinant porcine amelogenin and fluoride.

Recently, we used native amelogenins extracted from developing pig enamel to examine the combined effect of fluoride and amelogenins on the growth of octacalcium phosphate (OCP) and apatite crystals. The purpose of the present study was to investigate this combined effect using a highly purified recombinant amelogenin. We applied porcine amelogenin (rP172) and fluoride in a dual-membrane system as a model for tooth enamel formation. The combination of rP172 and fluoride in this system resulted in the formation of rod-like apatite crystals. On the other hand, without fluoride, rod-like OCP crystals of a comparable size were formed, and rather large hexagonal prisms of mixed crystals of OCP and apatite grew without amelogenins. Thus, highly purified and homogeneous recombinant amelogenin, in co-operation with F, regulated the mineral phase, habit, and size of crystals in the same manner as the extracted heterogeneous porcine amelogenins. We suggest that in both cases the control over the crystal phase and morphology was a direct effect of amelogenin protein serving as a scaffold for apatite mineralization.

Amelogenin↗

Calcium apatite crystals in synovial fluid rice bodies.

BACKGROUND: Rice bodies can occur in the joints in many rheumatic conditions, but they are most common in rheumatoid arthritis. They are generally believed to occur rarely in patients with osteoarthritis, but one study reported rice bodies with apatite crystals. OBJECTIVE: To report on a series of joint fluids with rice bodies containing apatite clumps and examine their clinical pictures. METHODS: All synovial fluid analysis reports for 10 years were reviewed for rice bodies and eight patients were reported on. A series of patients with a variety of diseases with synovial fluid rice bodies found to contain calcific material is described. All were examined by compensated polarised light and alizarin red stain, and four were examined by electron microscopy. RESULTS: The eight patients all had alizarin red S chunks embedded throughout the rice body. Transmission electron microscopy disclosed the presence of a matrix of collagen, fibrin, and amorphous materials containing typical apatite crystals. Clinical diagnoses, radiographic findings, and leucocyte counts varied, but six of the eight patients had had previous repeated corticosteroid injections into the joints. CONCLUSION: Aggregates of apatites may be more common than previously recognised in rice bodies as they are not routinely sought. Whether they are a result of joint damage or depot steroid injections and whether that might contribute to further joint injury now needs to be investigated.

Anthraquinones↗

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↗

Effect of salivary components on dissolution rates of carbonated apatites.

Salivary pellicle has been reported to retard the acid-induced demineralization of dental enamel. However, this effect has not been quantified, nor has the direct protective effect of pellicle on carbonated apatite dissolution been elucidated. We have developed a model capable of further studying these questions. Well-characterized synthetic ceramic carbonated apatite discs were pretreated with parotid or whole salivary secretions for 18 h or 7 days and then challenged with 0.1 mol/l acetate, pH 5.0, in a dissolution apparatus. Surface dissolution rates were compared to those from the same carbonated apatites with no saliva pretreatment. Pellicles formed from whole and parotid saliva in vitro and whole saliva in vivo were capable of reducing surface dissolution rates of ceramic carbonated apatites by up to 83% but the degree of this reduction was dependent on the type of saliva and the conditions of pretreatment.

Adsorption↗

Enamel apatite: chemistry, structure and properties.

The crystal chemistry of enamel apatite is probably the best understood of all the biological apatites. It is an imperfect apatite, low in calcium and hydroxide ions, but rich in substitutional impurities. A few impurities, like fluoride, are beneficial but most appear to disrupt the lattice structure. In this paper, the composition and basic structural features of enamel apatite are described, with particular emphasis on what we presently know, and do not know, about the role impurities play in establishing the fine details of this structure.

Apatites↗

The laser-Raman spectroscopic studies on human enamel and precipitated carbonate-containing apatites.

Laser-Raman spectra of synthetic apatites containing from zero to 12.4 wt% carbonate ions and enamel apatite are reported. Special attention is paid to the correlation between the intensities of the carbonate, phosphate, and hydroxyl modes and the carbonate content in the apatites. The data presented here indicate that most of the CO2-3 incorporated in the apatite lattice substitutes for PO3-4 and suggest that a small amount of CO2-3 might be located in the OH- position.

Apatites↗

Paracrystalline disorder of biological and synthetic carbonate-substituted apatites.

Paracrystalline disorder, a type of lattice imperfection which results in a loss of long-range crystalline order, was assessed in a range of biological and synthetic apatites using x-ray diffraction data. Carbonate substitution in the apatite lattice increased the amount of paracrystalline disorder in synthetic precipitated apatites, whereas trace amounts of fluoride and, to a lesser extent, zinc and strontium partially offset the disorder induced by carbonate. Paracrystalline disorder was significantly reduced in synthetic apatites prepared by high-temperature solid-state reaction. Human dental enamel was less disordered than either dentin or bone.

Apatites↗

Mineral acquisition rates in developing enamel on maxillary and mandibular incisors of rats and mice: implications to extracellular acid loading as apatite crystals mature.

UNLABELLED: The formation rates of mineral in developing enamel were determined by microweighing of incisors of mice and rats. Computations indicated that a large excess of hydrogen ions would result from creating apatite at the calculated rates. Enamel organ cells (ameloblasts), therefore, likely excrete bicarbonate ions to prevent pH in fluid bathing enamel from becoming too acidic. INTRODUCTION: Protons (H+) are generated whenever calcium and phosphate ions combine directly from aqueous solutions to form hydroxyapatite. Enamel is susceptible to potential acid loading during development because the amount of fluid bathing this tissue is small and its buffering capacity is low. The epithelial cells covering this tissue are also believed to form permeability barriers at times during the maturation stage when crystals grow at their fastest rates. The goal of this study was to measure the bulk weight of mineral present in rodent enamel at specific times in development and estimate the amount of acid potentially formed as the apatite crystals mature. MATERIALS AND METHODS: Upper and lower jaws of mice and rats were freeze-dried, and the enamel layers on the incisors were partitioned into a series of 0.5 mm (mouse) or 1.0 mm (rat) strips along the length of each tooth. The strips were weighed on a microbalance, ashed at 575 degrees C for 18-24 h to remove organic material, and reweighed to determine the actual mineral weight for each strip. RESULTS AND CONCLUSIONS: The data indicated that, despite differences in gross sizes and shapes of maxillary and mandibular incisors in rats and mice, the overall pattern and rates of mineral acquisition were remarkably similar. This included sharply increasing rates of mineral acquisition between the secretory and maturation stages, with peak levels approaching 40 microg/mm tooth length. Computer modeling indicated that quantities of H+ ions potentially generated as apatite crystals grew during the maturation stage greatly exceeded local buffering capacity of enamel fluid and matrix proteins. In other systems, bicarbonate ions are excreted to neutralize highly acidic materials generated extracellularly. Data from this study indicate that ameloblasts, and perhaps cells in other apatite-based hard tissues, use similar bicarbonate release mechanisms to control excess acid arising from mineral formation.

Ameloblasts↗