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Hydrophobicity, surface tension, and zeta potential measurements of glass-reinforced hydroxyapatite composites.

Wettability and zeta potential studies were performed to characterize the hydrophobicity, surface tension, and surface charge of P2O5-glass-reinforced hydroxyapatite composites. Quantitative phase analysis was performed by the Rietveld method using GSAS software applied to X-ray diffractograms. Surface charge was assessed by zeta potential measurements. Protein adsorption studies were performed using vitronectin. Contact angles and surface tensions variation with time were determined by the sessile and pendent drop techniques, respectively, using ADSA-P software. The highest (-18.1 mV) and lowest (-28.7 mV) values of zeta potential were found for hydroxyapatite (HA) and beta-tricalcium phosphate (beta-TCP), respectively, with composite materials presenting values in between. All studied bioceramic materials showed similar solid surface tension. For HA and beta-TCP, solid surface tensions of 46.7 and 45.3 mJ/m2, respectively, were obtained, while composites presented intermediate surface tension values. The dispersive component of surface tension was the predominant one for all materials studied. Adhesion work values between the vitronectin solution and HA and beta-TCP were found to be 79.8 and 88.0 mJ/m2, respectively, while the 4.0 wt % glass composites showed slightly lower values than the 2.5 wt % ones. The presence of beta-TCP influenced surface charge, hydrophobicity, and protein adsorption of the glass-reinforced HA composites, and therefore indirectly affected cell-biomaterial interactions.

Adsorption↗

Hydroxyapatite pseudopodagra. A syndrome of young women.

Six women with acute calcific periarthritis of the first metatarsophalangeal joint (hydroxyapatite pseudopodagra) are described, and 10 previously reported cases (8 female patients and 2 male patients) are reviewed. The onset was characterized by acute pain, swelling, erythema, tenderness, and limitation of movement of the first metatarsophalangeal joint, symptoms indistinguishable from those of gouty podagra, which is associated with transient amorphous calcific deposits in the vicinity of the joint. Based on these results it appears that hydroxyapatite pseudopodagra is a disorder that predominantly affects premenopausal women.

Acute Disease↗

Analysis of DNA structure by hydroxyapatite columns and ethidium bromide fluorescence techniques. A comparative study and effect on DNA binding.

Seven duplex DNA preparations have been structurally analyzed by hydroxyapatite column chromatography and an ethidium bromide fluorescence technique. Significant contamination of one preparation with single-stranded DNA was detected by hydroxyapatite column chromatography. Five of the other six preparations were found to contain significant single-stranded regions by the ethidium bromide fluorescence technique. Synthetic poly dAT was found to be duplex in structure. The presence of single-stranded regions considerably influenced DNA binding results in a radioimmunoassay.

Antibodies↗

"Milwaukee shoulder"--association of microspheroids containing hydroxyapatite crystals, active collagenase, and neutral protease with rotator cuff defects. II. Synovial fluid studies.

Hydroxyapatite crystals in spheroid-shaped masses 1.9-15.6 micrometer in diameter were found in 12 of 13 synovial fluids obtained from the shoulder joints of 4 patients with rotator cuff tears and glenohumeral osteoarthritis. Two of 16 control joint fluids also showed these particles. Collagen types I, II and III were identified in the joint fluid pellets from 3 of the 4 patients, and fibers with typical collagen periodicity were also seen on transmission electronmicroscopy. Collagenase and neutral protease activities were found in fluids from 5 joints in 3 patients, whereas active collagenase was found in only 1 of 10 fluids from rheumatoid arthritis patients and in none of 3 fluids from patients with osteoarthritis. Neutral protease activities were present in several rheumatoid joint fluids. These findings are compatible with the hypothesis of an enzymatic release of hydroxyapatite crystals from the synovium and endocytosis by synovial macrophage-like cells with subsequent crystal-stimulated release of collagenase and neutral protease into the joint fluid, completing a pathogenetic cycle.

Aged↗

"Milwaukee shoulder"--association of microspheroids containing hydroxyapatite crystals, active collagenase, ad neutral protease with rotator cuff defects. III. Morphologic and biochemical studies of an excised synovium showing chondromatosis.

Synovial tissue excised from the unstable right shoulder joint of a patient with an absent rotator cuff, severe glenohumeral joint degeneration, and hydroxyapatite-containing microspherules, collagen types I, II, and III, active collagenase, and neutral protease in the joint fluid showed extensive osteochondromatosis histologically. Electronmicroscopy revealed calcific foci in microvilli which could easily escape into the adjacent joint space through areas denuded of synovial cells. Fibrocytes demonstrated intensive pinocytotic activity of unknown significance. Energy dispersive analysis showed elemental ratios consistent with hydroxyapatite. A literature review suggested some relationships between the various pathologic lesions present in this joint. Whether similar synovial changes exist in the opposite shoulder joint of this patient and in 3 other subjects with nearly identical clinical, radiographic, and joint fluid findings is not known.

Aged↗

Hydroxyapatite deposition in osteoarthritic articular cartilage of the proximal femoral head.

Hydroxyapatite crystal deposition in the articular cartilage of the proximal femoral head was examined by using light and electron microscopy and electron probe microanalysis. The hydroxyapatite deposition was significantly more frequent in patients with osteoarthritis (40.7%, 22 of 54) than in control subjects (6.0%, 3 of 50) (P less than 0.001). Initial deposition of needle-like crystals was always seen on or within the electron-dense, amorphous material around the degenerating hypertrophic chondrocytes. Matrix vesicles were rarely seen in the calcifying areas.

Cartilage, Articular↗

Hydroxyapatite-coated strain gauges for long-term in vivo bone strain measurements.

The aim of this study was to examine the bonding process between hydroxyapatite-coated strain gauges and bone in order to continue development of a long term in vivo strain measurement device. Two types of commercially available hydroxyapatite (HA) particles were applied to the sensing surface of uniaxial strain gauges using a polysulfone solution as an adhesive. Characterization by scanning electron microscopy and x-ray diffraction (XRD) was used to determine materials property differences between the two powders. Interfacial strengths between the HA coatings and the strain gauges were tested and found comparable to interfacial strength obtained by a plasma sprayed HA coating on the surface of a titanium implant. Gauges were surgically placed on the periosteal surface of greyhound femora. Three groups of dogs were implanted with gauges for periods of 3, 6, and 12 weeks using cyanoacrylate, resorbable sutures, and cable ties to initially hold the gauge against the surface of the bone. Following euthanasia, the femora of the dogs were explanted and subjected to cantilever loading. Response of the implanted HA-coated gauges were compared to a control set that had been freshly glued onto the contralateral femur. Full response, that is, 100% of the strain measurement with respect to the control, was obtained after 12 weeks in vivo. Attachment of HA-coated gauges with circumferential suture showed bonding, while HA-coated gauges attached with cyanoacrylate did not bond to bone. After mechanical testing, femora were embedded in polymethylmethacrylate, cut, ground, and polished. Sections were stained using mineralized bone stain (MIBS) and optical microscopy was performed using transmitted and fluorescent light to allow analysis of remodeling occurring in the region of the strain gauges. Bone formation occurred at the HA surface of sutured gauges, and a fibrous tissue layer developed between the bone and HA coating when the tissue adhesive was used to initially bond the gauge. Fluorescence microscopy indicated an increase in the number of areas of bone remodeling adjacent to the gauge but a normal rate of remodeling of 0.93 +/- 0.07 mum/day was observed. No gross bone remodeling due to strain gauge placement was observed. Backscattered electron imaging (BSE) indicated new bone apposition at all time periods.

Animals↗

Evaluation of nanostructured carbonated hydroxyapatite coatings formed by a hybrid process of plasma spraying and hydrothermal synthesis.

Carbonated hydroxyapatite (CHA) coatings on a titanium alloy were prepared by hydrothermal synthesis of precursors plasma-sprayed with brushite as a raw powder. The structures, residual stresses, and bond strengths of the precursors and CHA coatings were investigated. The results showed that the sprayed precursors consisted of beta-Ca(2)P(2)O(7), alpha-Ca(3)(PO(4))(2), and CaHPO(4), whereas the CHA coatings exhibited a unique phase construction, nanostructured and needle-like crystals, and a fairly low tensile residual stress. The bond strength of a CHA coating 200 microm thick was 15 MPa, equivalent to that of a plasma-sprayed hydroxyapatite (HA) coating. The evaluation of the CHA coatings was performed together with that of plasma-sprayed HA coatings immersed in distilled water. The dissolution and bond-strength degradation of the CHA coatings were much lower than those of the plasma-sprayed HA coatings.

Carbon↗

In vivo evaluation of hydroxyapatite foams.

Hydroxyapatite (HA) is widely applied as bone graft material due to its osteoconductive potential and well-established biocompatibility. In this work, macroporous hydroxyapatite structures made through foaming of aqueous suspensions and gelcasting were tested for in vivo osteointegration. These foams are composed of a three-dimensional array of spherical pores with diameters of approximately 100-500 microm, interconnected by windows of smaller size in the range of 30-120 microm. The HA foams were implanted in the tibia of albino New Zealand rabbits and removed after a period of 8 weeks. Histological analysis revealed that the pores in the foams were partially or completely filled progressively with mature new bone tissue and osteoid after the implanted period. No immune or inflammatory reactions were detected. The high osteoconductive potential of the HA foams provides a potential structure for use as bone substitute in orthopedic, oral, and cranio-maxillofacial reconstructive surgery, and as dento-alveolar implants.

Animals↗

Delivery of insulin from hydroxyapatite ceramic microspheres: preliminary in vivo studies.

The major limiting factor in utilizing protein drugs for sustained delivery is the lack of suitable delivery systems. Ceramic hydroxyapatite microspheres are biocompatible and utilized for the purification of proteins. As a preliminary study, we have investigated the possibility of using hydroxyapatite ceramic microspheres loaded with insulin as an implantable delivery system in rats. With this limited test, it was shown that the loaded insulin was active and able to suppress the blood glucose level in normal rats.

Animals↗

Biological reactivity of zirconia-hydroxyapatite composites.

Materials and devices intended for end-use applications as implants and medical devices must be evaluated to determine their biocompatibility potential in contact with physiological systems. The use of standard practices of biological testing provides a reasonable level of confidence concerning the response of a living organism to a given material or device, as well as guidance in selecting the proper procedures to be carried out for the screening of new or modified materials. This article presents results from cytotoxicity assays of cell culture, skin irritation, and acute toxicity by systemic and intracutaneous injections for powders, ceramic bodies, and extract liquids of hydroxyapatite (HA), calcia partially stabilized zirconia (ZO), and two types of zirconia-hydroxyapatite composites (Z4H6 and Z6H4) with potential for future use as orthopedic and dental implants. They indicate that these materials present potential for this type of application because they meet the requirements of the standard practices recommended for evaluating the biological reactivity of ATCC cell cultures (CCL1 NCTC clone 929 of mouse connective tissue and CCL 81 of monkey connective tissue) and animals (rabbit and mouse) with direct or indirect patient contact, or by the injection of specific extracts prepared from the material under test. In addition, studies involving short-term intramuscular and long-term implantation assays to estimate the reaction of living tissue to the composites studied, and investigations on long-term effects that these materials can cause on the cellular metabolism, are already in progress.

Animals↗

Novel hydroxyapatite ceramics with an interconnective porous structure exhibit superior osteoconduction in vivo.

Calcium hydroxyapatite ceramics (CHA) are nontoxic materials, provoke little reaction from tissues, and by virtue of these properties represent a good starting point for creating bone substitutes. Although several porous CHAs have been used clinically, there have been few reports that CHA is fully replaced by newly formed bone, which may be due to its structure and the limited connectivity between pores. We recently developed a fully interconnected porous CHA (IP-CHA) by adopting a "foam-gel" technique. Structural analysis by scanning electron microscopy revealed that IP-CHA had spherical pores of uniform size that were interconnected by window-like holes. The surface of the wall structure was smooth, and hydroxyapatite particles were bound tightly to one another. Most of the interpore connections of IP-CHA ranged from 10 to 80 microm in diameter (average, 40 microm). When the cylindrical IP-CHA (diameter, 6 mm; height, 15 mm) was implanted into a rabbit femoral condyle, bone, and bone marrow with abundant vessels formed deep in the pores through the interpore connections. Within a period of 6 weeks, new bone had formed and penetrated to a distance of 3 mm from the surface of the IP-CHA implant. Furthermore, a compression test at 9 weeks revealed that the implanted IP-CHA steadily increased in strength to more than double the value of the initial test. These results indicate that the IP-CHA may have clinical utility as a superior bone substitute.

Animals↗

Effect of hydroxyapatite impregnation on skeletal bonding of porous coated implants.

Skeletal fixation of permanent implants by new methods such as fixation by mechanical interlocking of bone with porous prosthetic coatings or chemical bonding with bioactive materials shows growing potential. This paper reports on the resulting skeletal fixation of a combined porous and bioactive material. Metal plugs with a porous metal fiber coating impregnated with hydroxyapatite were implanted for 2, 4, and 12 weeks and were compared to the parent porous, nonbioactive, metal fiber material. Statistical analysis of the interfacial failure shear stress, as obtained by mechanical testing, shows there is a marked influence of hydroxyapatite impregnation on the rate of bone ingrowth and the strength of the interfacial bond the few weeks following surgery. Microscopical examination reveals that the apparent stimulation of bone ingrowth into the surface pores of the implant is the reason for the increased rate of bond formation. The results are of particular clinical interest: with an increased rate of bone ingrowth, weight bearing might be allowed much earlier, thus reducing the recuperation period.

Animals↗

Compressive properties of polymer coated synthetic hydroxyapatite for bone grafting.

A porous hydroxyapatite material hydrothermally converted from the calcium carbonate exoskeleton of the coral, genus Goniopora (CHAG) was either microcoated using polymethylmethacrylate (PMMA) or polylactic acid (PLA) to cover all internal surfaces, or externally coated to produce a shell, with the objective of reducing the brittleness of the material. Compressive testing showed that while CHAG, externally coated with PMMA, showed the largest increases in strength, stiffness, and energy absorption, the uncoated hydroxyapatite core cracked at low loads although the shell remained intact. CHAG internally microcoated with PMMA showed a 3.84 increase in compressive strength while specimens with internal PLA coatings had 1.81 times the compressive strength of uncoated CHAG specimens. Compared to the mechanical properties of cancellous graft material, specimens internally microcoated with either polymer could be produced having properties equivalent to or greater than those of cancellous graft.

Animals↗

Bone formation process in porous calcium carbonate and hydroxyapatite.

This study determined the bone formation in porous calcium carbonate (CC) and porous hydroxyapatite (HA) in ectopic sites. The bone formation stimulus was derived from bone marrow cells. CC and HA in the shape of disks were implanted with or without rat marrow cells into subcutaneous sites of syngeneic rats. The CC and HA had identical microstructure: pore size was 190-230 microns, porosity was 50-60% and they were fully interconnected. Bone did not form in any implants without marrow cells (disks themselves), whereas bone consistently formed in the pores of all implants with marrow cells after 4 weeks. The bone formation of both CC and HA occurred initially on surface of the pore regions and progressed toward the center of the pore. Scanning electron microscopy and electron-probe microanalysis revealed a continuum of calcium at the interfaces of both bone/CC and bone/HA implants. These results indicate that the bone formation in calcium carbonate derived from marine corals is comparable to the bioactive hydroxyapatite.

Animals↗

Protein interaction with tantalum: changes with oxide layer and hydroxyapatite at the interface.

For metallic implants the surface nature is extremely important because blood and tissue interactions with metal depend upon it. Protein adsorption is the initial reaction that takes place when an implant comes in contact with blood or tissue. We attempted to coat different thicknesses of oxide layers and hydroxyapatite on tantalum and examined the changes in water contact angle and adsorption of albumin and fibrinogen. Protein adsorption studies were performed with 125I-labeled proteins. A decrease in water contact angle was observed as the oxide layer thickness of tantalum increased. Fibrinogen adsorption increased on oxide layer coated and hydroxyapatite coated surfaces, compared to bare tantalum.

Adsorption↗

The ultrastructure of the bone-hydroxyapatite interface in vitro.

Rat bone marrow cells were cultured on plasma-sprayed hydroxyapatite (HA). The cells formed a mineralized extracellular matrix (ECM) that exhibited several characteristics of bone tissue. The interface between this mineralized ECM and the HA was studied at the ultrastructural level with scanning and transmission electron microscopy and x-ray microanalysis. Initially, the deposition of a globular, afibrillar matrix was observed on HA. This was followed by the integration of collagen fibers in this matrix and their subsequent mineralization. At the bone-HA interface two distinctly different interfacial structures were observed. An electron-dense layer with a thickness of 20-60 nm was regularly present, which contained both organic and inorganic material and was rich in glycosaminoglycans. The interfaces differed however, in the presence or absence of an amorphous zone which was free of collagen fibers and had an average thickness of 0.7-0.8 microns. It was frequently seen interposed between the electron-dense layer and the hydroxyapatite. Similar interfacial structures have also been described in the in vivo environment, where they were referred to as lamina limitans-like or cement linelike. From the results of this study, it can be concluded that the described in vitro system is a suitable model to study bone-biomaterial interactions.

Animals↗

Particle microelectrophoresis of calcium-deficient hydroxyapatite: solution composition and kinetic effects.

Concurrent work demonstrates that the zeta potential of bone is multivalued and systematically alterable by changes in sample preparation, steeping fluid composition, and steeping time. Since bone mineral is a mixture of carbonated calcium-deficient hydroxyapatites, and since the zeta potential of calcium-deficient hydroxyapatite (CDHA) is altered by pH and time in HNO3-KOH solutions, the zeta potential of CDHA in physiologic Neuman's fluid (NF) compared with that seen in bone could reveal important information on the contribution of the mineral phase to the zeta potential of bone. In addition, such information may be valuable in designing and evaluating calcium-phosphate ceramics for increased bone ingrowth. Results demonstrate that the zeta potential of CDHA in NF is negative. With increasing calcium in NF, the zeta potential magnitude of CDHA decreases and inverts to positive values given sufficient calcium concentration and steeping time. This result is opposite to that seen in bone, suggesting that exposed CDHA is not the predominant bone microsurface and implicating a bone surface protein component. With increasing phosphate in NF, the zeta potential magnitude increases to more negative values. While low concentrations of fluoride showed no effect, the possibility of an effect with higher concentrations is still to be determined.

Analysis of Variance↗