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Characterization of bone materials as ultrasonic transducers.

Physical, dielectric, piezoelectric and electromechanical parameters have been reported for bone and its two major constituents i.e. collagen and apatite, for their characterization as ultrasonic transducer materials. Collagen and apatite have been extracted from full bone using well known methods. These materials have been used to prepare simple disc-shaped test pieces (dimensions: diameter 10 mm and thickness 1.0 +/- 0.01 mm). The variation of various electrical parameters with frequency in the region (1-108 MHz) is examined for these materials. These include impedance, phase angle, relative voltage output, quality factor 'Q', dielectric constant and resistivity. The data so obtained are compared with those for ceramic and quartz transducers. The observations on impedance, phase and relative voltage output for bone materials indicate that the first resonance peak falls around 56 MHz followed by second and third harmonics around 112 MHz and 168 MHz respectively. A low 'Q' value suggests a fairly wide band transducer, while other parameters compare favourably with ceramic and quartz materials. Bone material has also been used to obtain a transducer in the standard configuration and velocity dispersion in the frequency range 1-108 MHz is examined.

Animals↗

Effects of hexafluorosilicate on the precipitate composition and dentine tubule occlusion by calcium phosphate.

OBJECTIVES: To improve the calcium phosphate precipitation (CPP) method for the occlusion of dentine tubules with calcium phosphate, the addition of calcium hexafluorosilicate (CaSiF6) to CPP solution was evaluated in vitro with respect to its occluding capacity and the composition of the precipitate. METHODS: The occlusion of dentine tubules was evaluated by SEM observations and by measurements of dentine permeability. The composition of the precipitate was determined by measuring the calcium to phosphorus (Ca/P) ratio of the precipitate in dentine tubules by energy dispersive X-ray spectroscopy (EDS). RESULTS: The addition of CaSiF6 to the CPP solution resulted in an increase of the Ca/P ratio in the precipitate not only on the dentine surface but also inside the dentine tubules; indicating that the precipitate became more apatitic in nature. The addition of CaSiF6 had no effect on occluding capacity in terms of measurements of dentine permeability or SEM observations. Dentine permeability decreased to approximately 4% of pretreatment values and the dentine tubules were occluded for approximately 10-15 microns from the dentine surface. CONCLUSION: It is concluded that the addition of CaSiF6 to CPP solution was desirable, since it provided a more apatitic precipitate in the dentine tubules, not only on the dentine surface but also inside the tubules, and there were no drawbacks with respect to its occluding capacity.

Apatites↗

Crystallographic identification of a calcium deposit in calcified pericarditis associated with articular chondrocalcinosis.

In a case of CPPD crystal deposition disease of the pseudorheumatoid type and of long duration, calcified constrictive pericarditis developed and was surgically treated. Analysis of the calcium deposit in the pericardium was carefully made by infrared absorption, x-ray diffraction, and thermogravimetry. It revealed that the deposit was composed of B-type carbonated apatite. Previously, both calcium pyrophosphate dihydrate (CPPD) and apatite crystals, either in the same place or in different tissues, have been reported in the same patient. These observations raise the possibility that the same metabolic error might lead to both types of crystal deposition.

Apatites↗

Bioactive calcium pyrophosphate glasses and glass-ceramics.

Calcium phosphate glass-based materials in the pyrophosphate region are briefly reviewed. Calcium pyrophosphate glasses can be prepared by including a small amount of TiO(2) (<or=10mol%). Bonelike apatite forms on some of the glasses in simulated body fluid. By heating powder-compacts of the glasses, they are crystallized and subsequently are sintered, resulting in fabrication of high-strength glass-ceramics with machinability; they are easier to be machined using conventional tools in comparison with conventional calcium phosphate ceramics. beta-Ca(2)P(2)O(7) crystal formed in the glass-ceramics plays an important role in the machinability. Their apatite-forming ability in simulated body fluid is drastically enhanced after autoclaving in distilled water. The glass-ceramics can be easily coated on a new beta-type titanium alloy using a conventional glazing technique.

Alloys↗

Fluorine-substituted hydroxyapatite scaffolds hydrothermally grown from aragonitic cuttlefish bones.

Porous hydroxyapatite scaffolds with different levels of fluorine substitution (46% and 85%) on the OH sites were produced via hydrothermal transformation of aragonitic cuttlefish bones at 200 degrees C and calcination at temperatures up to 1200 degrees C. The increasing level of F substitution reduces the kinetics and probably the yield of the reaction. The incorporation of F in the lattice of hydroxypatite caused a lowering of the unit cell volume due to reduction of the length of the a-axis. The crystallites formed were close in size to bone-like apatite and were orientated along the a-axis rather than the c-axis. There was evidence of AB-type carbonated apatite.

Animals↗

The in vitro bioactivity of two novel hydrophilic, partially degradable bone cements.

Composite bone cements were prepared with bioactive glasses (MgO-SiO(2)-3CaO.P(2)O(5)) of different reactivities. The matrix of these so-called hydrophilic, partially degradable and bioactive cements was composed of a starch/cellulose acetate blend and poly(2-hydroxyethyl methacrylate). The addition of 30 wt.% of glasses to this system made them bioactive in acellular medium: a dense apatite layer formed on the surface after 7 days of immersion in simulated body fluid. This was demonstrated both by microscopic and infrared spectroscopic techniques. The composition of the glass and, consequently, its structure was found to have important effects on the rate of the apatite formation. The combination of reactivity obtained by one formulation with the hydrophilic and degradable character of these cements makes them a very promising alternative to conventional acrylic bone cements, by allowing a better stabilization of the implant and a stronger adhesion to the bone.

Apatites↗

Sintered hydroxyfluorapatites. Part III: sintering and resultant mechanical properties of sintered blends of hydroxyapatite and fluorapatite.

The presence of chemically dissimilar apatites occurs widely in biological systems as a result of exposure to chemically enriched media, but also to optimize the mechanical properties. The use of mechanical blends of apatites can be used to manufacture high temperature processed fluoride containing hydroxyapatites with improved properties. Mechanical blends of fluorapatite and hydroxyapatite were produced with 0%, 20%, 40%, 60%, 80% and 100% fluorapatite. Pellets were sintered at 1150 degrees C, 1200 degrees C and 1250 degrees C and the density determined by the Archimedes method. Mechanical properties including hardness, elastic modulus and fracture toughness were measured using indentation. It was found that mechanical blends of 150 nm sized hydroxyapatite and 300 nm sized fluorapatite lead to solid solutions after sintering. The mechanical blends do not sinter as effectively as homogeneous hydroxyfluorapatite solid solutions and exhibit a minimum density at 80 wt.% fluorapatite. The hardness, elastic modulus and brittleness decreases with a higher flourapatite content, attributed primarily to a decrease in density. The higher fracture toughness for mechanical blends indicates that these materials are more crack resistant and provide a means for improving mechanical properties.

Apatites↗

Sintered hydroxyfluorapatites--IV: The effect of fluoride substitutions upon colonisation of hydroxyapatites by mouse embryonic stem cells.

Biodegradable scaffolds serve a central role for tissue engineering scaffolds and guiding tissue regeneration. Some of these scaffolds, including apatites, display a significant effect upon cell adhesion and cell proliferation. The incorporation of scaffold technology with the developing embryonic stem (ES) cell field and the capacity of ES cells for self-renewal and differentiation are believed to hold enormous potential for applications in biomedical research and regenerative medicine. The purpose of this work was to determine the effect of hydroxyapatite (HAP) and fluoride substitutions of HAP upon ES cell growth and colonisation. Sintered hydroxyfluorapatite discs were found to support cellular proliferation and colonisation, and the ES cells displayed a tendency for differentiation on the apatite surface as determined by reductions in colony Oct4 immunoreactivity. Fluoride-containing HAPs were found to provide equivalent support to gelatin in terms of cell numbers, yet superior support for cellular colonisation when compared to HAP. This study indicates that fluoride substitutions of HAP may represent a viable strategy for the development of certain engineered tissue replacements and tissue regeneration systems using ES cells.

Analysis of Variance↗

Stability and cellular responses to fluorapatite-collagen composites.

Fluorapatite (FA)-collagen composites were synthesized via a biomimetic coprecipitation method in order to improve the structural stability and cellular responses. Different amounts of ammonium fluoride (NH4F), acting as a fluorine source for FA, were added to the precipitation of the composites. The precipitated composites were freeze-dried and isostatically pressed in a dense body. The added fluorine was incorporated nearly fully into the apatite structure (fluoridation), and a near stoichiometric FA-collagen composite was obtained with complete fluoridation. The freeze-dried composites had a typical biomimetic network, consisting of collagen fibers and precipitates of nano-sized apatite crystals. The human osteoblast-like cells on the FA-collagen composites exhibited significantly higher proliferation and differentiation (according to alkaline phosphatase activity) than those on the hydroxyapatite-collagen composite. These enhanced osteoblastic cell responses were attributed to the fluorine release and the reduced dissolution rate.

Alkaline Phosphatase↗

SEM and TEM study of the hierarchical structure of C57BL/6J and C3H/HeJ mice trabecular bone.

Transmission electron microscopy (TEM) and scanning electron microscopy (SEM) were used to study the hierarchical structure of trabecular bone from C57BL/6J (low bone mass) and C3H/HeJ mice (high bone mass). Bone was harvested from two different anatomical locations: femoral metaphysis and L5 vertebra. This investigation focused on three structural scales: the mesostructural (porous network of trabecular struts), the microstructural (collagen fibril arrangements in trabecular packets), and the nanostructural (collagen fibril and apatite crystals) levels. At the mesostructural level, no distinct differences were found in the trabecular structure of femoral metaphysis but thinner trabecular struts were observed in L5 vertebra for C57BL/6J mice strain. At the microstructural level, the collagen fibrils forming the rotated, twisted, and orthogonal plywood arrangements were distinguished as well as atypical arrangements. At the nanostructural level, the shape and size of apatite crystals, and their arrangement with respect to collagen fibrils were studied. In spite of very different bone mass densities, both mice strains had similar structures at the nanostructural and microstructural levels.

Animals↗

Comparison of ceramic hydroxy- and fluoroapatite versus Protein A/G-based resins in the isolation of a recombinant human antibody from cell culture supernatant.

A recombinant human antibody (IgG(1)-subtype) was produced in Chinese Hamster Ovary (CHO) cells. Alternatives to the established isolation by Protein A affinity chromatography were investigated. Neither an alternative elution agent (Arginine) nor an alternative affinity ligand (Protein G) resulted in an improvement in yield and/or purity. Subsequently, apatite stationary phases including a novel ceramic fluoroapatite material were tested. By applying a double gradient (first 0 to 1M NaCl, then 0.01 to 0.4M phosphate) the culture supernatant was separated into three fractions: the flow through, which contained no active antibody, the NaCl-eluate, which contained the antibody and no other discernible protein contaminants, and a fraction that eluted in the phosphate gradient and contained several proteins, but no active antibody. In case of the hydroxyapatite, retention of the antibody decreased and yield increased when the pH was raised from 6.0 to 8.2 (isoelectric point (pI) of the antibody: 8.3), to reach a yield of 71% at pH of 8.2. In case of the fluoroapatite, retention was also found to increase with increasing mobile phase pH, but the yields went through a maximum (of ca. 90%) at a mobile phase pH of 7.0. No traces of contaminants were seen in the corresponding gel. This is the first time that yields of 90% and such high purities have been reported as the result of a single chromatographic step for the antibody in question with either (Protein A) affinity or apatite chromatography.

Antibodies↗

Cyclic fatigue in water of veneer-framework composites for all-ceramic dental bridges.

OBJECTIVES: Ceramic materials applied in dentistry may exhibit significant subcritical crack growth due to the severe cyclic loading in the aqueous environment encountered in the mouth during mastication. The authors report on the subcritical crack growth behavior of three dental restoration systems (Empress 2/IPS Eris, TZP/Cercon S and Inceram-Zirconia/Vita VM7) under cyclic loading in water, in order to establish guidelines for the use and design of long-lifetime all-ceramic posterior bridges. METHODS: Inert strength and lifetime tests under cyclic loading in an aqueous environment were performed in a mechanical bending apparatus and evaluated with Weibull statistics. RESULTS: Subcritical crack growth occurred predominantly in the outer veneer layer of the veneer-framework composites. The apatite-based veneer (IPS Eris) was more susceptible to subcritical crack propagation compared to the feldspathic glass veneers (Cercon S and Vita VM7). SIGNIFICANCE: Dental restoration systems containing apatite-based veneers and weak frameworks (Empress 2/IPS Eris) are not recommended for the fabrication of all-ceramic bridges in the molar region. Conversely, veneer-framework systems consisting of feldspathic glass veneers and tough zirconia-based frameworks (TZP/Cercon S and Inceram-Zirconia/Vita VM7) may exhibit lifetimes longer than 20 years if the bridge connector is properly designed.

Aluminum Oxide↗

Characterization of sintered TCP sheets with various contents of binder prepared by tape-casting technique.

OBJECTIVES: The purpose of this study was to investigate the effect of binder content on the characterization of sintered tricalcium phosphate (TCP) sheets prepared by the tape-casting technique. METHODS: Poly(acrylic acid)-based binder was added at four contents: 5, 10, 15, or 30 mass%, to a beta-TCP aqueous slurry. The TCP green sheets were fabricated with the prepared TCP slurry by the tape-casting technique, and were sintered at 1200 degrees C for 2h. The sintered TCP sheets were characterized by field-emission scanning electron microscope (FE-SEM) observation, X-ray diffraction (XRD) and Fourier transform infrared (FT-IR) spectrometry. Apatite precipitation on the sintered TCP sheet surface in Hanks' balanced salt (HBS) solution at pH 7.4 was monitored over 14 days at 37 degrees C. RESULTS: A TCP green sheet with 5 mass% binder content had insufficient flexibility before sintering, and that with 30 mass% binder content had a problem in controlling the viscosity of slurry. FE-SEM observation showed that no differences in grain size existed in the various sintered TCP sheets. Measurement of XRD and FT-IR showed the presence of alpha-TCP phase in the various sintered TCP sheets, and same crystallographic structure was observed among various sintered TCP sheets. After 14 days immersion in HBS solution, the surfaces of sintered TCP sheets were completely covered with precipitated apatite crystals. SIGNIFICANCE: The binder content of 10 or 15 mass% is suitable for sintering TCP sheets. The sintered TCP sheet prepared by the tape-casting technique is a promising material as bone substitute.

Acrylic Resins↗

Saturation of human saliva with respect to calcium salts.

It may be assumed that free ionic concentrations of calcium and phosphate in resting saliva tend to equilibrate with those in plaque fluid, and that salivary data can therefore be used to illustrate chemical conditions in both saliva and plaque. In the present study, salivary data collected from the literature or obtained in our laboratory were used to calculate degrees of super- and undersaturation with respect to apatites, brushite, beta-tricalcium phosphate, octacalcium phosphate, calcium carbonate and calcium fluoride in the pH range from 3 to 9. Concentrations of calcium, phosphate, fluoride, carbonate, and background ion strength of resting parotid saliva, resting submandibular saliva, and resting and stimulated whole saliva were entered into a computer program, and curves illustrating saturation in the pH range 3-9 constructed. It was found that oral fluids are supersaturated with respect to apatites above pH 5.3 and with respect to octacalcium phosphate and beta-tricalcium phosphate above pH 6. Parotid saliva was undersaturated with respect to brushite whilst submandibular saliva was supersaturated with respect to that salt in the pH range 6-8. Stimulated whole saliva with 25 mmol/l carbonate became supersaturated with respect to calcium carbonate only above pH 7.3, which may explain the absence of this salt in the human oral cavity. To maintain the saturation of oral fluids with respect to calcium fluoride, i.e. to ensure its survival in the mouth required 6 ppm fluoride in the aqueous phase. Therefore, this salt, the outcome of topical fluoride therapy, will inevitably dissolve in the oral fluids.

Adult↗

A strategy of cystine stone management.

Simple mechanical disintegration of cystine calculi by extracorporeal shock wave lithotripsy and/or percutaneous nephrolithotomy is being performed widely around the world. However, it cannot be denied that the cystine calculus is one of the most difficult stones for mechanical disintegration. We previously reported the oral medical chemolysis of cystine calculi in 1982 and a third of the patients became stone-free. In another third of the patients the cystine component was replaced by apatite during the medical treatment and apatite stones formed in this manner are easily disintegrated. In view of the complications of mechanical disintegration, oral medical treatment for chemolysis should be recommended before simple destruction of cystine calculi.

Adult↗

Urolithiasis in rats with diabetes insipidus (Brattleboro strain rats).

Brattleboro strain rats homozygous for hypothalamic diabetes insipidus (DI rat) excrete nearly their body weight per day in dilute urine and yet can form bladders stones composed of struvite and apatite. Studies were undertaken to investigate this apparent paradox. The results show that DI rat urine is indeed undersaturated with respect to struvite and apatite. However, chronic infection of DI rat urine with a urease-containing organism (Proteus mirabilis) results in the rapid formation of large struvite/apatite bladder stones. It is concluded that the Brattleboro strain DI rat, like man and unlike other rats, forms struvite/apatite calculi only in the presence at chronic urinary tract infection.

Animals↗

An optimized synthetic substrate for orthodontic bond strength testing.

OBJECTIVE: The purpose of this study was to investigate the suitability of different synthetic calcium phosphate based substrates as a biomimetic enamel surface model for orthodontic bond strength testing. METHODS: Carbonated apatite, amorphous calcium phosphate and commercial hydroxyapatite specimens were prepared as substrates for orthodontic bond strength testing. Carbonated apatite specimens were prepared by pressing, sintering and treatment with NaF. The shear bond strength was measured with a universal testing machine. RESULTS: Hydroxyapatite, amorphous and cold pressed carbonated hydroxyapatite exhibited fractures within the substrate after debonding. Mean bond strength values for carbonated hydroxyapatite were 7.38 (1.75) MPa for specimens pressed at 300 degrees C and 9.55 (2.23) MPa for specimens pressed at 300 degrees C and then sintered at 600 degrees C. An additional NaF treatment after sintering resulted in lower bond strength measurements of on average 6.52 (1.03) MPa. SIGNIFICANCE: Hot pressed and sintered carbonated hydroxyapatite showed acceptable shear bond strength values and may represent a suitable biomimetic model for orthodontic bond strength testing.

Apatites↗

An X-ray diffraction study of the effects of heat treatment on bone mineral microstructure.

A series of human cortical bone specimens has been heated to temperatures up to 1200 degrees C and the mineral content examined in detail by X-ray diffraction. Line profile analysis of the diffraction data has been undertaken to characterise the microstructural (crystallite size and microstrain) features of the mineral at each temperature. Individual profile fitting of several maxima from each diffractogram has also provided precise lattice parameters of the apatite at each temperature. The apatite did not show any significant decomposition over the temperature range although CaO was increasingly formed at temperatures above 600 degrees C. Both finite crystallite size and microstrain contributed significantly to the diffraction peak broadening below 600 degrees C. When heated to > 800 degrees C, the small, rod-like mineral crystallites changed from a highly anisotropically strained state to one with significantly larger equidimensional crystals possessing little microstrain. The findings are discussed in the context of graft bone substitutes and surgical heating of bone.

Adult↗