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Induction of bioactivity of a non-bioactive glass-ceramic by a chemical treatment.

Glass-ceramic A-W(Al), which was prepared by heat treatment of a MgO-CaO-SiO2-P2O5-Al2O3 glass to precipitate crystalline apatite and wollastonite, shows a higher mechanical strength than glass-ceramic A-W, which was prepared by heat treatment of a MgO-CaO-SiO2-P2O5 glass to precipitate the same types of crystalline phases. The former, however, does not show bone-bonding ability, i.e. bioactivity, whereas the latter shows it. In the present study, in order to induce bioactivity of glass-ceramic A-W(Al), it was treated with HCl or NaOH solutions with different concentrations, and its bioactivity was evaluated by examining the apatite formation on its surface in a simulated body fluid (SBF) with ion concentrations nearly equal to those of human blood plasma. When the glass-ceramic A-W(Al) was pretreated with HCl aqueous solutions with concentrations over 0.1 M, it formed the bone-like apatite on its surface in SBF. This was attributed to the formation of a hydrated silica on its surface by the HCl treatment.

Apatites↗

Morphology and microstructure of electrochemically deposited calcium phosphates in a modified simulated body fluid.

Calcium phosphates were deposited on a pure titanium plate for various loading times under 1.3 and 12.9 mA/cm2 in a modified simulated body fluid at 52-92 degrees C. The plate-like crystals formed under 1.3 mA/cm2 were identified to be octacalcium phosphate (OCP) and/or carbonate-containing apatite. OCP converted to carbonate-containing apatite with the elapsing time of the loading current. The needle-like precipitates formed under 12.9 mA/cm2 were identified to be carbonate-containing apatite crystals elongated parallel to the c-axis direction. The degrees of crystallinity of the deposits formed under 1.3 mA/cm2 showed the highest value around 72 degrees C, whereas those under 12.9 mA/cm2 increased with the electrolyte temperature. The carbonate content of the deposits decreased with the electrolyte temperature and the flow current. The degree of crystallinity of the electrochemically deposited calcium phosphates decreased with the carbonate content.

Apatites↗

Calcium-level responsive controlled drug delivery from implant dosage forms to treat osteoporosis in an animal model.

The effects of plasma calcium levels on estradiol release from a self-setting apatite bone cement containing 0.5% estradiol and on the bone mineral density (BMD) of ovariectomized rats were investigated. Apatite cement consisting of an equimolar mixture of tetracalcium phosphate, dicalcium phosphate dihydrate and 0.5% beta-estradiol was prepared. The in vitro release profiles from the cements in simulated body fluid containing 0, 5 and 10 mg/100 ml calcium indicated that estradiol release rate decreased with increasing calcium concentration in the dissolution medium. After subcutaneous implantation of the cement, in vivo estradiol release in diseased rats (ovariectomized rats on a low calcium diet) was significantly higher than that in normal rats. The diseased rats maintained a low calcium level during drug release. The bone mass of the recovery model rat was greater after the experiment than before. The results suggested that the severity of osteoporosis in this animals can be reduced by the implantation of this estradiol-loaded apatite cement.

Animals↗

Complementary information on bone ultrastructure from scanning small angle X-ray scattering and Fourier-transform infrared microspectroscopy.

Scanning small angle X-ray scattering (scanning SAXS) and Fourier-transform infrared microspectroscopy (FT-IRM) have previously been utilized independently to characterize the structural properties of bone in an anatomical position-resolved fashion. Whereas SAXS provides a direct measure of the physical characteristics of apatitic crystals, FT-IRM assesses structure of both mineral and organic matrix at the molecular level. In the present study both methods were applied to examine the same developing bone tissue from the L-4 vertebra of a 14-month-old (accidental death). A 200-microm-thick section was processed for examination by scanning electron microscopy and SAXS. Spectra were collected at 200 microm spatial resolution at specific locations in cortical and cancellous bone. Parameters determined included total SAXS intensity, crystal thickness (T), and degree and direction of predominant crystal orientation. For FT-IRM analysis, a section 4 microm thick was cut longitudinally from the top of the sample. Spectra of regions 100 x 100 microm2 were acquired from the same locations as the SAXS spectra. Integrated areas of the phosphate nu(1,3) collagen amide I, and carbonate nu2 absorbances, were calculated to obtain mineral: matrix and carbonate:mineral ratios. The relative quantities of types A, B, and labile carbonate (substituted for apatite hydroxyl, phosphate, and surface positions, respectively) were also evaluated. Polarized FT-IRM data were collected to determine molecular orientation of the apatite and collagen components. The results of this study show that the information obtained from the two techniques is complementary. Both SAXS and FT-IRM data revealed that the crystals were significantly larger in the cancellous region compared with the cortical region, that mineralization was greater in the cortex, and that the crystals were oriented to a larger degree in the cancellous compared with the cortical bone. The scanning SAXS measure of crystal thickness was significantly correlated to the FT-IRM measures of crystallinity, type A carbonate substitution, and crystal orientation. In conclusion, it was found that the combined use of SAXS and FT-IRM provides valuable, unique information on structural changes in bone at both the microstructural and ultrastructural level. Although each method can be used individually, the combination of techniques provides additional insights into the mechanism of bone crystal maturation.

Apatites↗

Suffocation of nerve fibers by living nanovesicles: a model simulation--part II.

Nanobacteria may cause peripheral neuropathy by adhesion to the perineurium. This hypothesis receives support from five independent observations: (1) identification of perineurial apatite in diabetic patients with peripheral neuropathy, (2) massive presence of nanobacteria in a diabetic patient, (3) beneficial effect of lasers on peripheral neuropathy, (4) model simulation indicating that perineurial deposition and attachment of nanobacteria is encouraged by both their size and chemical nature, and (5) transient inhibition of neural function by apatite. Initial deposition of (stressed) nanobacteria is promoted by a slime thought to consist of proteins, calcium, and phosphate, and is most likely followed by an immobilization phase, mediated by a bioadhesive capacity of the apatite. Proteomics may hold the key to control both attachment processes.

Apatites↗

Pre-mineralisation of starch/polycrapolactone bone tissue engineering scaffolds by a calcium-silicate-based process.

This work describes a new methodology to produce bioactive coatings on the surface of starch-based biodegradable polymers or other degradable polymeric biomaterials. As an alternative to the more typical bioactive glass precursors, a calcium silicate gel is being employed as a nucleating agent, for inducing the biomimetic formation of a calcium-phosphate (Ca-P) layer. The method has the advantage of being able to coat efficiently both compact materials and porous 3-D architectures aimed at being used on tissue replacement applications and as bone tissue engineering scaffolds. This treatment is also very effective in reducing the incubation periods, being possible to observe the formation of an apatite-like layer, only after 12 h of immersion in a simulated body fluid (SBF). The apatite coatings formed on the compact surfaces or along the fibres of a fibre mesh scaffold structure made from a starch/polycrapolactone blend (SPCL) were analysed and compared in terms of morphology, chemical composition and structure. After the first days of SBF immersion, the apatite-like films exhibit the typical cauliflower like morphology. With increasing immersion times, these films exhibited a partially amorphous nature and the Ca/P ratios became very closer to the value attributed to hydroxyapatite (1.67). It was possible to fully pre-mineralise the SPCL scaffolds and simultaneously to keep the porous morphology of the fibre-bonded scaffold.

Absorption↗

The biomimetics of bone: engineered glass-ceramics a paradigm for in vitro biomineralization studies.

In this study, we investigated the behavior of fetal rat osteoblasts cultured up to 23 days on a bioactive apatite-wollastonite glass-ceramic (AW) and on the same material on which a carbonated apatite layer was formed by a biomimetic process (AWa). The specific activity of alkaline phosphatase activity was about 30% increased on AWa compared to AW disks at the last day of culture. Scanning electron microscopic (SEM) observations of the material surfaces after scrapping off the cell layers revealed that mineralized bone nodules remained attached to both surfaces but in larger numbers on AWa. The AWa/bone interfaces were also analyzed after fracturing the disks and by transmission electron microscopy (TEM). All these results indicated the importance of the surface composition in supporting differentiation of osteogenic cells and the subsequent apposition of bone matrix. Furthermore, prefabrication of a biological apatite layer by a biomimetic method could improve our knowledge of biomineralization processes and could find application as bone-repairing material.

Alkaline Phosphatase↗

Metabolic evaluation of infected renal lithiasis: clinical relevance.

Complete metabolic evaluation was performed in 21 patients with infected renal lithiasis. Patients with pure struvite stones (struvite +/- carbonate apatite) were significantly less likely to have metabolic abnormalities than patients who had struvite +/- carbonate apatite+calcium oxalate (2 of 14 v 7 of 7, P = 0.0003). Urine calcium excretion was markedly higher in the mixed stone group than the pure struvite group (342 +/- 98 mg/24 h v 136 +/- 82 mg/24 h; P < 0.0001). The differing opinions among researchers regarding the likelihood of finding metabolic abnormalities in patients with urolithiasis and infection probably reflect differences in the definitions of the populations studied. If patients with calculi containing only struvite +/- carbonate apatite are evaluated, we believe that few significant metabolic abnormalities will be identified.

Adolescent↗

Ultrastructure and pathogenesis of human urinary calculi.

The ultrastructure of human urinary calculi was studied using scanning and transmission electron microscopy. The hydroxyapatite constituent of the stones was often present in the form of sperical aggregates of the minute apatite crystallites (1 to 10 mu in diameter). In most cases, the sperical apatite deposits consisted of concentric lamellae of crystallites. The spherical apatite deposits, described in detail for the first time in urolithiasis, were similar to those found in a variety of calcified tissues including nephrocalcinosis and malakoplakia.

Apatites↗

Crystal-associated nephropathy in patients with brushite nephrolithiasis.

BACKGROUND: We have biopsied the renal cortex and papillae of patients who form brushite renal stones asking if this unusual stone type is associated with specific tissue changes. We contrasted these with biopsies of 15 calcium oxalate stone formers, three stone formers with intestinal bypass, and four normal subjects. METHODS: We studied all ten brushite stone formers treated with percutaneous nephrolithotomy (PNL) during the past 3 years using digital video imaging of renal papillae, and obtained cortical and papillary biopsies. Biopsies were analyzed by light and electron microscopy, microinfrared spectroscopy, and electron diffraction. RESULTS: Apatite crystals plugged scattered terminal collecting ducts whose cells were injured or dead, and surrounding interstitium inflamed and fibrotic. White papillary deposits of interstitial apatite particles, so called Randall's plaque, were also present. Glomerular changes and cortical tubular atrophy and interstitial fibrosis were moderate to severe. CONCLUSION: Brushite stone formers combine the interstitial plaque of calcium oxalate stone formers with the collecting duct apatite plugs found in stone formers with intestinal bypass. Collecting duct injury and interstitial fibrosis are severe. Prominent cortical fibrosis, tubule atrophy, and glomerular pathology seem secondary to the collecting duct plugging. We believe crystallization obstructs and destroys terminal collecting duct segments thereby damaging nephrons, perhaps via intranephronal obstruction, and producing a hitherto unrecognized renal disease.

Adolescent↗

Crystalline structure of salivary calculi. A microradiographic and microdiffractometric study.

The crystalline substances localized in different areas of salivary calculi were identified by using microradiographic and X-ray microdiffractometric techniques. All 19 calculi analyzed had a general pattern of apatite which was recognizable both in the center and at varying distances from the core. In addition to apatite patterns, six of the calculi gave a diffraction pattern of whitlockite, which in all but one case was located in the central part of the calculi. The influence of various factors on the distribution of the crystalline substances of apatite and whitlockite within the calculi is discussed.

Apatites↗

Radiocarbon: analysis of inorganic carbon of fossil bone and enamel.

Carbon dioxide from calcium carbonate in fossil bone can be selectively separated from carbon dioxide in bone apatite by hydrolyzing the sample first in acetic acid and then in hydrochloric acid. Radiocarbon analyses of the inorganic carbon dioxide in three samples of known age clearly show calcium carbonate in fossil bone to be secondary and the carbonate of bone apatite to be indigenous and suitable for dating in some cases. Agreement between dates on collagen-bone apatite pairs increases the level of confidence.

Acetates↗

Enamel erosion by some soft drinks and orange juices relative to their pH, buffering effect and contents of calcium phosphate.

The capability of a soft drink or a juice to erode dental enamel depends not only on the pH of the drink, but also on its buffering effect. As the latter is the ability of the drink to resist a change of pH it may add to the effects of the actual pH. The aim of the present study was to compare the pH and the buffering effect of various soft drinks with their erosive effects and the solubility of apatite. In 18 soft drinks, mineral waters and juices available on the Danish market, pH and the concentrations of calcium, phosphate and fluoride were determined. The buffering effect was determined by titration with NaOH. Human teeth (n = 54) covered with nail varnish except for 3x4-mm windows were exposed to 1.5 liters of the drink for either 7 days or 24 h under constant agitation. The depth of the erosions was assessed in longitudinal sections. The depth was found to vary greatly from 3 mm eroded by the most acidic drinks and fresh orange juice to only slightly affected surfaces by most of the mineral waters. The dissolution of enamel increased logarithmically inversely with the pH of the drink and parallel with the solubility of enamel apatite. Orange juice, pH 4.0, supplemented with 40 mmol/l calcium and 30 mmol/l phosphate did not erode the enamel as the calcium and phosphate saturated the drink with respect to apatite. Generally, the lower the pH the more NaOH was necessary to bring the pH to neutrality. In particular the buffering effect of the juice was high. For all drinks, no effect of their low fluoride concentrations was observed.

Apatites↗

Chemolitholysis and lithotripsy of infectious urinary stones - an in vitro study.

OBJECTIVES: This study was performed to look for an improvement of therapeutic strategies with regard to the treatment of infectious urinary stones using artificial stones made of struvite and apatite ('Bon(n) stones') which are comparable to their natural counterparts. MATERIALS AND METHODS: Using an experimental arrangement simulating the physiological conditions in the upper urinary tract, the efficacy of artificial urine (pH 5.7), Suby G solution (pH 3.6), mixtures of artificial urine with Suby G (pH 3.9 and pH 4.1) in dissolving artificial struvite and apatite stones (Bon(n) stones) was investigated. The dissolution of natural infectious urinary stones was also measured. Additionally, investigations on shock-wave lithotripsy (SWL) combined with initial chemolytic treatment of the stones were performed. RESULTS: The efficacy of Suby G solution in dissolving artificial stones was demonstrated. Direct comparison of chemolysis of natural and artificial stones showed no statistical difference between infectious urinary stones and Bon(n) stones of the same material. The investigations on SWL showed a significant improvement on stone comminution, especially of artificial apatite stones after initial chemolytic treatment with Suby G. CONCLUSION: New basics to improve dissolution of infectious urinary stones have been developed by performing standardized in vitro investigations. Local chemolysis with Suby G is an effective tool in the treatment of infectious stone disease. SWL can be improved by varying the physical properties of infectious stones through initial treatment with Suby G solution.

Apatites↗

Defluoridation of drinking water by boiling with brushite and calcite.

Existing methods for defluoridating drinking water involve expensive high technology or are slow, inefficient and/or unhygienic. A new method is now suggested, encompassing brushite and calcite suspension followed by boiling. Our aim was to examine the efficiency of the method and the chemical reactions involved. Brushite, 0.3-0.5 g, and an equal weight of calcite were suspended in 1 litre water containing 5-20 ppm fluoride. The suspensions were boiled in an electric kettle, left to cool and the calcium salts to sediment. Solution ion concentrations were determined and sediments were examined by X-ray diffraction. In distilled water initially containing 5, 10 and 20 ppm fluoride the concentration was reduced to 0.06, 0.4 and 5.9 ppm, respectively. Using Aarhus tap water which contained 2.6 mmol/l calcium the final concentrations were 1.2, 2.5 and 7.7 ppm, respectively, and runs without calcite gave results similar to those with calcite. Without boiling the fluoride concentration remained unaltered, as did the brushite and calcite salts, despite occasional agitation by hand. All solutions were supersaturated with respect to fluorapatite and hydroxyapatite and close to saturation with respect to brushite. Boiling produced well-crystallised apatite and traces of calcite, while boiling of brushite alone left a poorly crystallised apatite. We conclude that boiling a brushite/calcite suspension rapidly converts the two salts to apatite which incorporates fluoride if present in solution, and that this process may be exploited to defluoridate drinking water.

Apatites↗

In vitro caries-like lesion formation in F-containing tooth enamel.

In vitro caries-like lesion formation in F-containing tooth enamel (shark) showed a lower depth of penetration compared to that in human enamel (42 vs. 100%). After longer periods of exposure, the depth of penetration in shark enamel is only 33% relative to that in human enamel. The extent of dissolution in acetate buffer for powdered human enamel and for synthetic F-free calcium-carbonated-apatites was greater than for powdered shark enamel and for synthetic F-containing apatites. These results suggest that the cariostatic action of fluoride may be explained in terms of a combination of two effects: (a) the stabilizing effect against acid dissolution (caries) when fluoride is incorporated in the tooth mineral (apatite), and (b) a greater degree of remineralization (repair) in the presence of F solution.

Animals↗

Hydrolysis of dicalcium phosphate dihydrate in the presence or absence of calcium fluoride.

Effects of temperature (25 and 37 degrees C), pH (4.9-10.5), and CaF2 on CaHPO4.2H2O (DCPD) hydrolysis were studied in a pH-stat. Octa-calcium phosphate (OCP) was the product at pH 6.2-6.8 and 25-37 degrees C; thermodynamically stable apatitic compounds were formed at higher pH and/or higher temperature. In the presence of CaF2, apatite was the product, its crystallinity improved, and the fluoride content increased as pH of the reaction decreased. The results demonstrate the remarkable ability of fluoride to promote the hydrolysis of an acidic calcium phosphate, DCPD, to apatite.

Apatites↗

Amelogenin-guided crystal growth on fluoroapatite glass-ceramics.

The formation of aligned fibrous apatite crystals in enamel is predominantly attributed to the involvement of amelogenin proteins. We developed a model to study interactions of matrix proteins with apatite mineral in vitro and tested the hypothesis that amelogenin solubility affects the ability to induce protein-guided mineralization. Crystal growth experiments were performed on fluoroapatite (FAP) glass-ceramics in mineralizing solutions containing recombinant full-length amelogenin (rH174) at different concentrations. Using atomic force microscopy, we observed that mineral precipitated randomly on the substrate, but also formed thin layers (height, 10 nm) on FAP within 24 hrs. This growth pattern was unaffected when 0.4 mg/mL of rH174 was added. In contrast, crystals grew on FAP at a rate up to 20 times higher, at 1.6 mg/mL protein. Furthermore, nanospheres and mineral bound specifically to FAP and aligned in strings approximately parallel to the c-axis of FAP, leading us to the conclusion that amelogenin proteins indeed control direction and rate of growth of apatite in enamel.

Amelogenin↗