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Apatite formation on collagen fibrils in the presence of polyacrylic acid.

Recently, we reported the formation of bone-like apatite on collagen fibrils by biomimetic method. Compounds containing carboxyl moieties are believed to be effective in the formation of apatite. Polyacrylic acid [-CH2CH(COOH)-]n (PAAc) is widely used in dentistry. In the present study, the effect of PAAc in the formation of apatite from revised simulated body fluid (R-SBF) on collagen fibrils was studied. Two different experimental approaches were tried to study the effect of PAAc present in the collagen and in the R-SBF solution. In the first, collagen gel was soaked with 1 mg/ml PAAc (molecular weights 25000 and 100000) for the time intervals of 30 min and 6 h. The gels were then dried in air and incubated in R-SBF. Characterization of the precipitates formed on the collagen fibrils in gel showed that the formation of apatite was inhibited when soaked in PAAc for 6 h. In the second experiment, when PAAc (0.1 and 1.0 mg/ml) was mixed with R-SBF the microstructure of the precipitates formed on collagen fibrils was affected partially. Hence, it is confirmed that the presence of PAAc in the biomimetic environment of collagen affects the mineralization of apatite.

Acrylic Resins↗

Apatite-forming ability of alginate fibers treated with calcium hydroxide solution.

Calcium alginate fibers were prepared by extruding an aqueous sodium alginate solution into an aqueous calcium chloride solution. The fibers were treated with a saturated aqueous calcium hydroxide solution for various periods and their apatite-forming ability was examined in a simulated body fluid (SBF). The calcium alginate fibers were treated with the aqueous calcium hydroxide solution for periods longer than five days formed apatite on their surfaces in SBF, and their apatite-forming ability improved with increasing calcium hydroxide treatment time. The amount of calcium ions released from the fibers also increased with increasing calcium hydroxide treatment time, resulting in acceleration of nucleation and growth of apatite on the fiber surfaces. The resultant apatite-alginate fiber composite is expected to be useful as a flexible bioactive bone-repairing material.

Alginates↗

Uremia-related vascular calcification: more than apatite deposition.

In the present study, we characterized and compared the mineral phase deposited in the aortic wall of two different frequently used chronic renal failure rat models of vascular calcification. Vascular calcification was induced in rats by either a 4-week adenine treatment followed by a 10-week high-phosphate diet or 5/6 nephrectomy followed by 6 weeks of 0.25 microg/kg/day calcitriol treatment and a high-phosphate diet. Multi-element mapping for calcium and phosphate together with mineral identification was performed on several regions of aortic sections by means of synchrotron X-ray-mu-fluorescence and diffraction. Bulk calcium and magnesium content of the aorta was assessed using flame atomic absorption spectrometry. Based on the diffraction data the Von Kossa-positive precipitate in the aortic regions (N=38) could be classified into three groups: (1) amorphous precipitate (absence of any diffraction peak pattern, N=12); (2) apatite (N=16); (3) a combination of apatite and magnesium-containing whitlockite (N=10). The occurrence of these precipitates differed significantly between the two models. Furthermore, the combination of apatite and whitlockite was exclusively found in the calcitriol-treated animals. These data indicate that in adenine/phosphate-induced uremia-related vascular calcification, apatite is the main component of the mineral phase. The presence of magnesium-containing whitlockite found in addition to apatite in the vitamin D-treated rats, has to be seen in view of the well-known vitamin D-stimulated gastrointestinal absorption of magnesium.

Animals↗

[Clinical experience with the implantation of collagen-apatite for local bone regeneration].

After a short review of the problematic of bone substitution the possibility of bone regeneration induced by implantation of synthetic Apatite and reaffined heterogenic Collagen is displayed especially with an industrial manufactured mixed preparation called "Collapat". Firstly it is referred especially to the comparing animal experience and histological results by Nizard. According to this research fine dispersed Apatite powder has obviously a remarkable osteoinductive effect, which is inhenced by the Collagen component, used especially as a distribution medium. The tract mainly concerns the clinical experiences in human patients with Collagen-Apatite-implantation since October 1979 with the indication of larger bone defects. In 85 patients with totally 94 Collagen-Apatite-implantations in different age groups no incompatibility was seen. In X-ray controls a rapid reestablishment of natural bone structures was observed. The Collagen-Apatite product seems particularly to be apt for the reconstruction of defect cavities as well as for an additive to autologous bone transplantation which by this method can be limited as well in number as in quantity.

Adolescent↗

Solid-state phosphorus-31 nuclear magnetic resonance differentiation of bone mineral and synthetic apatite used to fill bone defects.

Bioabsorption of synthetic apatite compounds used to promote bone healing and remodeling has been difficult to evaluate. In this study, solid-state phosphorus-31 nuclear magnetic resonance (NMR) has been used to characterize and quantitate bone mineral and a synthetic apatite in order to establish a model for bioabsorption studies. Pulverized solid samples of cortical rabbit bone and a synthetic fluoridated apatite were examined in vitro at variable degrees of hydration. A 9.4 T superconducting spectrometer was used to obtain 31P magic angle spinning NMR spectra and T1 relaxation times. Quantitation was attempted in mixed samples using T1 recovery data. Bone mineral and synthetic apatite could be distinguished by chemical shift and T1 relaxation time in variable hydration states, and were readily differentiated in mixtures by their T1 relaxation time. NMR estimates of relative proportions of components in mixed samples were accurate within 2% of evaluations based on weight. Solid-state 31P NMR therefore provides a suitable method for monitoring the bioabsorption of synthetic apatites.

Animals↗

Synergistic effect of silanol group and calcium ion in chitosan membrane on apatite forming ability in simulated body fluid.

A chitosan membrane modified with silanol groups and calcium ions on its surface and in its structure, respectively, was newly developed and evaluated for the potential application as a bioactive-guided bone-regeneration membrane. The chitosan membrane, which contained calcium nitrate tetrahydrate, was prepared and further subjected to surface modification with 3-isocyanatopropyl triethoxysilane (IPTS) following hydrolysis with HCl solution. As control, chitosan membranes which contained only calcium nitrate tetrahydrate and modified with only silanol groups were prepared, respectively. Three membranes were exposed to simulated body fluid (SBF) for a period ranging from 3 h to 7 days. The SBF exposure led to the deposition of a layer of apatite crystals on the surface of the chitosan membrane modified with silanol groups and calcium ions, while those modified with only calcium ions or silanol groups did not show the apatite-forming ability. It implies that the silanol groups and calcium ion acted together in a synergistic fashion in the formation of apatite crystals; the silanol groups and calcium ions acted as the nucleation sites and accelerator for the formation of apatite crystals, respectively. Therefore, this new chitosan membrane is likely to have a potential for the application as a bioactive guided bone regeneration membrane because of its apatite-forming ability in the SBF.

Apatites↗

The effect of supersaturation on apatite crystal formation in aqueous solutions at physiologic pH and temperature.

The importance of supersaturation in the dynamics of apatite precipitation from aqueous solutions is well-established. To determine whether this parameter has a comparable impact on the concomitant development of the textural properties of this phase, such as crystal size and shape, we investigated mineral accretion in synthetic solutions seeded with 0.67 g/L apatite over a range of supersaturations at pH 7.4 and 37 degrees C. A dual specific-ion electrode-controlled titration method was used to maintain the seeded reactions under the following solution conditions: 1.0 to 1.8 mmol/L Ca2+, 0.67 to 1.2 mmol/L total phosphate (PO4), Ca/PO4 (initial) = 1.5, 143 mmol/L KNO3, and 10 mmol/L HEPES. Samples were collected for chemical and textural analyses when the seed apatite was reduced by new accretions to 1/2, 1/4, 1/8, 1/16, and 1/32 of the total solids in suspension. All new accretions were found to be apatitic. At the lowest supersaturation, accretion occurred primarily by growth of the seed crystals. However, at the highest supersaturation examined, the crystals at the end of the experiments were actually smaller, on average, than the original seeds, even though the total mass increased 32-fold. The results suggest that proliferation of new crystals supplanted growth of the seed crystals as supersaturation was increased. The results also suggest that differences in tissue fluid supersaturation may contribute to the large disparity in dimensions between dentin and enamel apatite crystals.

Apatites↗

Effect of apatite crystals on the activity of amelogenin degrading enzymes in vitro.

The objective of the present study was to determine the effect of apatite crystals on the activity of amelogenin degrading enzymes in vitro. Current experimental data, together with previous reports support the view that among the different proteinases present in the enamel extracellular matrix, serine proteinase(s) are responsible for the massive degradation of amelogenins during the maturation stage. For our in-vitro experiments we used the recombinant amelogenin M179 as substrate and a "65%-satd. (NH4)2SO4" fraction of enamel proteins as well as chymotrypsin as sources for serine-proteinase activity. We report preliminary experiments of amelogenin proteolysis in the presence of apatite crystals resulting in a different proteolysis pattern when compared to amelogenin proteolysis without apatite crystals. Quantitative analysis of the HPLC peaks corresponding to the proteolysis products indicates that the presence of apatite crystals in the proteolysis solution inhibits the ability of the serine-proteinases to degrade amelogenin. The present observations support the hypothesis that amelogenin degradation correlates with apatite crystal growth during enamel maturation.

Amelogenin↗

Biocompatibility of apatite ceramics in mandibles.

Apatite ceramics composed of synthetic hydroxyapatite, Ca10(PO4)6(OH)2 were implanted in mandibles of adults dogs. The histological observations indicated that the apatite ceramic was closely contacted with newly formed bone tissue without any rejection phenomenon up to 2 weeks. An electron micrograph of non-decalcified ultra-thin section at 8 weeks showed that the apatite ceramic was directly bonded to newly formed bone and the mineralized bone tissue was grown into the micro pores of the ceramics independent on the pore size. It seemed that the outer surface of the apatite was exchanged by bone tissue. From these results it was considered that the apatite ceramic was a successful implant material in dental and surgical fields.

Animals↗

In vivo effect of fluoride-substituted apatite on rat bone.

Different types of calcium phosphate compounds are commercially available for medical and dental applications as bone substitute materials. Biological apatites contain several kinds of minor elements such as carbonate (CO3), magnesium (Mg), and fluoride (F) in enamel, dentin, and bone. It has been shown that F ion and F-substituted apatite promoted osteoblast proliferation and inhibited osteoclast cell activity. The purpose of this study was to investigate the in vivo rat tibia activity on F-substituted apatite (FAp). Apatites of unsintered calcium deficient apatite (CDA), and FAps, with low, medium, and high F concentrations, were implanted in rat tibia for 1 and 2 weeks. Implanted tissues were embedded in paraffin blocks, stained with hematoxylin-eosin and histomorphometrically observed. Results showed that low F concentration induced better and faster new bone formation in vivo compared to CDA. Therefore the results suggested that F as a minor element in bone rendered a suitable effect on bone formation in vivo.

Animals↗

Apatite-type crystal deposition in arthritic cartilage.

Osteoarthritis or 'Joint Failure' is a multi-factorial disease with a final common pathway of cartilage degeneration and bone eburnation. The association of arthritic disease and joint degeneration with the deposition of sodium urate crystals in gout and calcium pyrophosphate crystals in pseudogout (chondrocalcinosis) is clinically well established. Electron microscopy coupled with electron probe analysis has revealed the presence of various other calcium phosphate crystals in joint tissues and fluids. We have found three new morphological types of apatite crystals in human articular cartilage which are too small to be detected by X-rays of human joints or even by light microscopy of joint tissues. Two morphologically distinct types of apatite crystals in articular cartilage are associated with extracellular matrix vesicles formed from the cell processes of chondrocytes. 'Cuboid' crystals, which are found in the pericellular regions near the surface zone of articular cartilage, appear to be a variant of apatite and may even be 'Whitlockite' because there are traces of magnesium present. There are increased numbers of these microscopic 'cuboid' crystals (Type II) and Mineral Nodules (Type I) in arthritic cartilage and this is coupled with increased numbers of matrix vesicles and alkaline phosphatase activity. Clusters of fine needle-shaped apatite crystals (Type III) found on the surface of articular cartilage are not associated with matrix vesicles. Thus some forms of osteoarthritis are closely associated with apatite type crystal deposition and may imply abnormal mineral formation in articular cartilage as a pathogenic mechanism.

Apatites↗

Crystallinity in apatites: how can a truly disordered fraction be distinguished from nanosize crystalline domains?

In the last decade synthetic apatites mimicking the human natural one have been widely prepared and characterized from the physico-chemical point of view; however a shading zone is still remaining related to the evaluation and distinction of the less crystalline part, almost amorphous, and the crystallographically well ordered, nano-sized part, inside the apatite itself. Actually natural apatite forming bone tissue can include both types of crystals whose prevalence is dependent from the specific bone evolution stage and the specialized tissue performance. The quantitative description of such a combination usually represents a puzzling problem, but the result can also clarify the definition of "crystallinity in apatite" that appears still controversial. Many different synthetic apatites, including those nucleated on organic templates, were analyzed with different techniques (X-ray diffraction, transmission electron microscopy, and so on) to clarify the true nature of the disordered part. The results, manipulated by the classical methodologies devised for substances with highly perturbed structural order, led to establish that only specifically prepared amorphous calcium phosphate is really a glass, while the distorted portion coexisting with more or less crystalline regions is simply nanocrystalline. Moreover, at the conceptual limit of crystallinity tending to zero, the two models surprisingly cease to be conflicting.

Calcium Phosphates↗

Mechanical and biological properties of apatite composite resins.

Well-crystallized hydroxyapatite was synthesized at 80 degrees C and pH 7.4, and mixed as filler with 2.2'(4-methacryloxydiethoxyphenyl) propane. BPO and DHPT were used as polymerization initiators. The compressive strength and Knoop hardness of the apatite composite resins increased with the increase of apatite content, and approached a plateau above an apatite-resin ratio (Ap/R) of 1. The thermal expansion coefficient of the composite with apatite-resin ratio Ap/R = 1 was almost equal to that of teeth. In tooth cavities, the composites seemed to adhere well to enamel without a bonding agent. The biocompatibility of the composites implanted hypodermically into rats appeared to improve with the increase of apatite content, and implants showed no significant inflammation after 1 wk and 4 wk.

Acrylates↗

Interaction between self-assembled protein vesicles and microporous apatite surface.

Self-assembled structures such as vesicles have generated immense interest in recent decades due to their potential in mimicking biological membranes and in acting as drug-delivery systems. Despite the importance of the interaction between these organized assemblies and the surface of biomaterials, little is known about the mechanism involved. In this study, the interaction between giant proline-rich mussel adhesive protein (MAP) vesicles and the microporous apatite surface was investigated by scanning electron microscopy (SEM). We have found that MAP vesicles incubated on the apatite surface similar to osteoclasts, induce site-specific resorption of the apatite surface. However, in contrast to the osteoclastic resorption, the vesicle-induced resorption process appears to be accompanied by an organic matrix-mediated remineralization process. This results in the formation of a variety of complex three-dimensional site-specific "remodelled zones" on the apatite surface of micrometre scale. The mechanism of the formation of "remodelled zones" is discussed in terms of surface phenomena, such as adsorption and deformation of vesicles, site-specific release of resorptive agents, organic matrix-directed remineralization, and Ca-induced fusion, collapse and reshaping of the vesicles on the apatite surface.

Journal Article↗

Effect of thermal treatment on apatite-forming ability of NaOH-treated tantalum metal.

The prerequisite for an artificial material to bond to living bone is the formation of bonelike apatite on its surface in the body. This apatite can be reproduced on its surface even in an acellular simulated body fluid with ion concentrations nearly equal to those of the human blood plasma. The present authors previously showed that the tantalum metal subjected to a NaOH treatment to form a sodium tantalate hydrogel layer on its surface forms the bonelike apatite on its surface in SBF in a short period. The gel layer as-formed on the metal is, however, not resistant against abrasion, and hence thus-treated metal is not useful for clinical applications. In the present study, effects of thermal treatment on the mechanical properties and apatite-forming ability of the NaOH-treated tantalum metal were investigated. The sodium tantalate gel on the NaOH-treated tantalum was dehydrated to convert into amorphous sodium tantalate by a thermal treatment at 300 degrees C in air environment and into crystalline sodium tantalates by the thermal treatment at 500 degrees C. Resistivity of the gel layer against both peeling-off and scratching was significantly improved by the thermal treatment at 300 degrees C. The high apatite-forming ability of the sodium tantalate hydrogel was a little decreased by the thermal treatment at 300 degrees C, but appreciably decreased by the thermal treatment at 500 degrees C. It is believed that the tantalum metal subjected to the 0.5 M-NaOH treatment and the subsequent thermal treatment at 300 degrees C is useful as implants in dental and orthopaedic fields, since it shows high bioactivity as well as high fracture toughness.

Journal Article↗

Preparation and characterization of fluoride-substituted apatites.

Apatites were prepared with three different fluoride concentrations: 0.0 mM (pure hydroxyapatite) 2.5 mM and 5 mM. Reactions were performed in aqueous medium using a reaction between diammonium orthophosphate and calcium nitrate 4-hydrate and ammonium fluoride at temperatures of 3 degrees, 25 degrees, 60 degrees and 90 degrees C. The effects of reaction temperature and fluoride concentration on the crystal morphology, phase purity and crystallinity of the precipitates were observed, using transmission electron microscopy (TEM), X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy and ion chromatography. Transmission electron micrographs revealed that the crystallites precipitated at 3 degrees C were spheroidal, but became increasingly acicular with increasing precipitation temperature. X-ray diffraction results indicated that all the materials produced were phase pure and that the crystallinity of apatites prepared at higher precipitation temperatures was higher than those prepared at lower precipitation temperatures. A significant difference in the a-axis dimension of fluoride-substituted apatites was observed, as compared to hydroxyapatite. FTIR spectroscopy revealed a hydroxyl band at 3568 cm-1, along with a broad peak of adsorbed water in the region of 3568 cm-1 to 2670 cm-1 in the hydroxyapatite and fluoride-substituted apatite powders. Hence by careful selection of the precipitation conditions and fluoride contents, the composition and morphology of fluoride-substituted apatite may be controlled and this has interesting implications for the development of these materials for biomedical implantation.

Journal Article↗

Synthesis and proposed crystal structure of a disordered cadmium arsenate apatite Cd5(AsO4)3Cl(1-2x-y)O(x)[symbol: see text](x)OH(y).

During a study into the synthesis of minerals composed of mining wastes aimed at improving their immobilisation, a cadmium arsenate apatite has been prepared by hydrothermal methods. The structure of this apatite was analysed by single crystal X-ray diffraction, and was found to consist of a standard apatite framework based on Cd(5)(AsO(4))(3)X, where X represents an anion resident on the (0,0,0.25) site. The framework is hexagonal with the space group P6(3)/m(no 176), a= 9.9709(8), c= 6.4916(4)[Angstrom]. The X ion site is predominantly occupied by Cl(-) ions; however due to significant shortening of the c axis exhibited by all cadmium containing apatite phases, a pure chlorapatite is not possible without a significant cation deficiency. No evidence of the necessary deficiency was found in the crystal structure. For larger bromo- and iodo-apatites significant modulations along the c-axis are required to accommodate the halide. This paper examines a number of compensation mechanisms and proposes that a minor disorder of chloride, oxide and hydroxide located on the X ion site provides the required charge compensation mechanism. This is contrary to previous complex modulations proposed in the literature. The proposed chemical formula is Cd(5)(AsO(4))(3)Cl(1-2x-y)O(x)[symbol:see text](x)OH(y) where [symbol: see text] represents a vacancy.

Journal Article↗

Evidence for Differential Unroofing in the Adirondack Mountains, New York State, Determined by Apatite Fission-Track Thermochronology.

Apatite fission-track ages of 168-83 Ma for 39 samples of Proterozoic crystalline rocks, three samples of Cambrian Potsdam sandstone, and one Cretaceous lamprophyre dike from the Adirondack Mountains in New York State indicate that unroofing in this region occurred from Late Jurassic through Early Cretaceous. Samples from the High Peaks section of the Adirondack massif yielded the oldest apatite fission-track ages (168-135 Ma), indicating that it was exhumed first. Unroofing along the northern, northwestern, and southwestern margins of the Adirondacks began slightly later, as shown by younger apatite fission-track ages (146-114 Ma) determined for these rocks. This delay in exhumation may have resulted from burial of the peripheral regions by sediment shed from the High Peaks. Apatite fission-track ages for samples from the southeastern Adirondacks are distinctly younger (112-83 Ma) than those determined for the rest of the Adirondack region. These younger apatite fission-track ages are from a section of the Adirondacks dissected by shear zones and post-Ordovician north-northeast-trending normal faults. Differential unroofing may have been accommodated by reactivation of the faults in a reverse sense of motion with maximum compressive stress, sigma1, oriented west-northwest. A change in the orientation of the post-Early Cretaceous paleostress field is supported by a change in the trend of Cretaceous lamprophyre dikes from east-west to west-northwest.

Journal Article↗