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PubMed1996

Carbonate hydroxyapatite gel monolith formation and drying.

The effect of carbonate in reducing the crystal size of precipitated hydroxyapatite by approximately an order of magnitude has not been used previously in the preparation of gel monoliths for the fabrication of carbonate hydroxyapatite ceramics. The aim of this study was to devise a method whereby gel monoliths of carbonate hydroxyapatite could be repeatably produced without cracking. A precipitation reaction was used for the preparation of carbonate hydroxyapatities with carbonate contents of 5.8 and 7.8 wt%. Biaxial vacuum filtration was used to form disc shaped monoliths. The rate of filtration of a 7.8 wt% carbonate hydroxyapatite sol was measured throughout the gelation process. Gel monoliths were dried slowly in air and the mass and dimensions of the gel were recorded once approximately every 24 hours. Using this data, the permeability, water volume fraction with time, rate of water loss, gelation point and gel density were determined. The pore size distribution was measured using mercury porosimetry for a carbonate apatite gel and a pressed powder pellet of a commercial hydroxyapatite. In tact monoliths were formed with masses up to 9.9 g. It was found that gelation behaviour was independent of monolith size and carbonate content and the final green density of all monoliths was 37%. Gelation was found to occur at 50-55 vol% water. Gel monoliths were found to have a monomodal pore size distribution with a mean pore size of 9.1 nm, whereas a pressed pellet of hydroxyapatite had a bimodal pore size distribution.

Apatites↗
PubMedDate not supplied

[Necessity and validity of standard models for experimental preclinical evaluation of biomaterials. An example of biologic characterization of a hydroxyapatite-based implant material].

A large number of methods are now available for the preclinical screening of implantable materials concerning their biocompatibility and their ability to stimulate tissue formation. In vitro techniques represent a very useful tool, since this way we can realistically simulate the biological events which occur in vivo at the bone-implant interface. In the present study scanning electron microscopy and light microscopy observations were performed in order to assess the effect of an hydroxyapatite granulate on cell behaviour and morphology. Uptake of proteins to hydroxyapatite surface has been also investigated by comparing the amounts adsorbed after incubation with bovine serum albumin and bovine pancreaticamilase. According to our preliminary observation cells do not show signs of toxicity or inhibition of cell growth even after 14 days of co-culture with hydroxyapatite. Granules were covered by an uninterrupted cell layer by day seven. Even after two days micrographs show cells anchored and spread over the surface of the underlying granules, with a flattened and stellate shape. Such a morphology indicates a very high cellular activity, suggesting that the interaction with hydroxyapatite seriously increased metabolism. Measurements of protein adsorption on the hydroxyapatite surface show that changes in the size of particles affect the binding of proteins, while, in the case of granular hydroxyapatite, despite changes in size of granules, variations of protein adsorption were not observed, neither in relation to their different isoelectric point. Our preliminary results represent a good example of the opportunities presented by an experimental in vitro model.

Biocompatible Materials↗
PubMed1997

Hydroxyapatite orbital implants--our local experience.

Hydroxyapatite orbital implants were first introduced by Arthur Perry in 1985 and approved by the Food and Drug Administration for general use in 1989. Since its inception, it has become a popular orbital implant as it combines biocompatibility with improved motility. At the Singapore National Eye Centre, twenty consecutive patients underwent hydroxyapatite implantation from June 1993 to June 1996. Seventeen patients who had a follow-up time of more than 6 months were analysed retrospectively. Sixteen had enucleation while one had evisceration performed. The implant size used was either 18 or 20 mm. Seven primary and 10 secondary hydroxyapatite implants were performed in 17 patients ranging in age from 20 to 65 years. Six consisted of coralline hydroxyapatite while the remaining 11 were made of artificial hydroxyapatite. In the follow-up time of 6 to 16 months (mean 10.2 months), there have been no cases of infection, extrusion or migration. Two patients who had implant exposure were managed surgically with fascia lata grafts and have healed well. It appears that hydroxyapatite is the implant of choice for an anophthalmic socket. It becomes incorporated by the patients own tissues and once vascularised, it is less likely to migrate or extrude. The complication rate is low and implant exposure, if it occurs, can be easily managed.

Adult↗
PubMed1997

[Morphological studies of hydroxyapatite crystals exposed to disodium pamidronate].

"In vitro" effects of disodium pamidronate on hydroxyapatite crystals morphology, and some "in vivo" data from bone powder of tibia and vertebrae from treated young and mature rabbits are here reported. Hydroxyapatite, synthesized following Rigoli et al method, and bone powder from rabbits were studied with X-ray, infrared and raman emission techniques for crystallographic analysis. Adsorption studies were also performed with a balanced solution of hydroxyapatite exposed to different times, 48, 120 and 168 hours and concentrations 1 x 10(-5) M, 3 x 10(-5) M, 8 x 10(-5) M y 1 x 10(-4) M of pamidronate. Infrared and raman spectrometry were not conclusive due to technical bias, but X-ray difractograms showed pure hydroxyapatite crystals in an hexagonal system. At constant time, pamidronate concentrations were varied, showing after 48 hours of exposition, a slight growth in the 002 plane, an aleatoric behavior in 213 and a marked increase in 004. After 120 hours, 002 plane is steady with a net growth in 004 and 213. After 168 hours, the 3 mentioned planes grow in proportion to pamidronate concentrations, tending to enlarge the crystal shape. Plane 13 markedly grow with pamidronate 8 x 10(-5) M a 1 x 10(-4) M, which are biologically high concentrations. Potentiometric assessments, in the 1 x 10(-5) to 1 x 10(-4) M range of concentrations show that bisphosphonate was completely adsorbed to the crystals. Additional "in vivo" observations showed changes in bone powder crystals isolated from pamidronate treated young animals, involving a growing of planes 002 and 211, in samples from both epiphysis and diaphysis, regarding untreated samples. Changes were more evident at epiphysis. In mature rabbits, it was shown a decrease in basal plane 002 and growing at 210, 211 and 310 with a trend to enlarge the crystal shape in diaphysis and to shorten it in vertebrate spongiosa. The "in vivo" doses are equivalent to those used by Ferretti et al. in intact rats with pamidronate low dose groups, showing an improvement of bone material properties and stiffness. Thus, it may rather be lower than the "in vitro" used concentrations. In concordance with above experimental conditions it can be concluded that bisphosphonates exert morphological changes in hydroxyapatite crystals, in a dose dependent manner, at least when high concentrations are used. In addition, it is postulated that changes observed on "in vivo" samples may be the result with other adaptative factors as for example the local mechanical usage. The latter data were limited, and should be studied with more details if an extrapolation to the bisphosphonate treated osteoporotic women is intended. Finally, it is suggested that any agent that changes BMU activity (all known anti-osteoporotic drugs) may potentially modify the quality of hydroxyapatite crystals, affecting in turn the bone resistance to fracture, independently from the quantity of bone mass gained. Thus, to help predicting the consequences on skeletal fragility, there is a need to know the direct or indirect effect of drugs on bone crystals.

Animals↗
PubMed1993

Hydroxyapatite in experimental laryngotracheal reconstruction.

Current concepts in laryngotracheal reconstruction include lumen augmentation with interposition of autogenous costal cartilage. In an effort to reduce potential increased morbidity at the donor site, the use of porous hydroxyapatite was compared with autogenous costal cartilage in an experimental study. Ninety-nine New Zealand rabbits were randomly operated on to receive autogenous costal cartilage (n = 32), hydroxyapatite covered by a perichondrium graft (n = 33), or hydroxyapatite alone (n = 34). Rabbits were killed at 3 months after the surgery. Clinically (n = 85), no difference was found among the groups. Histologically (n = 81), grafts (n = 54) in the group receiving autogenous costal cartilage were greater than in those receiving the hydroxyapatite. Concerning the value of epithelial layer covering the graft, the rate of inflammation, the cricoid-graft interface, and the graft viability, no difference was noted among the groups. In the group in which no graft was found (n = 27), a fibrous bridge maintaining a cricoid distraction was noted in 23 cases. This study shows the possible value of hydroxyapatite in the animal model, but before clinical application, the natural course of the fibrous bridge must be documented.

Animals↗
PubMed1993

Colocalization of cholesterol and hydroxyapatite in human atherosclerotic lesions.

Cholesterol and calcium phosphate, the latter in the form of hydroxyapatite, accumulate in atherosclerotic lesions. In this report, we demonstrate that these organic and inorganic constituents of lesions can accumulate together, closely associated in crystal agglomerates. Using the fluorescent cholesterol probe, filipin, we identified unesterified cholesterol that was associated with calcium granules in tissue sections of lesions. We also have shown that small crystallites of cholesterol can associate with preformed hydroxyapatite crystals in vitro. Scanning electron microscopy coupled with energy-dispersive X-ray analysis demonstrated the physical association of many small crystallites of cholesterol with larger crystals of hydroxyapatite. These small crystallites of cholesterol associated with hydroxyapatite stained with filipin. This contrasted with the lack of filipin staining of unassociated larger cholesterol crystals or hydroxyapatite alone. How cholesterol and calcium come to be closely associated in crystal agglomerates within atherosclerotic lesions remains to be determined.

Aorta↗
PubMed2005

[Influence of different mixing fluids on mechanical and micromorphological in vitro qualities of hydroxyapatite cement].

AIM: In reconstructive craniofacial surgery a new synthetic hydroxyapatite cement has been used in the last few years. Tetra- and dicalcium phosphates react with either slow setting aqua bidest. or with faster setting sodium monophosphate solution to hydroxyapatite. The aim of this study was to investigate the two mixing fluids of hydroxyapatite for resulting micromorphology, pressure strength, and interactions with fibroblasts. MATERIAL AND METHODS: Pressure strength tests of hydroxyapatite cement test samples (n=80) were performed after a setting time of 3 and 24 h. The micromorphology of surfaces of the resulting particles was assessed under an electron microscope. In cell cultures L-929 and HEp2 cells were incubated with test samples and cell growth was assessed by light and electron microscopy. RESULTS: The test samples mixed with sodium monophosphate solution showed statistically significantly higher values of pressure strength. The resulting pressure strength depended on respective mixing fluids, setting time, and pressure of application. In general, test samples mixed with sodium monophosphate solution showed a more load-stable, homogeneous anorganic matrix whereas test samples mixed with aqua bidest. had a porous microarchitecture which was more fragile. In cell culture the porous structure showed disintegration in cell culture medium. CONCLUSION: To what extent the two mixing liquids of hydroxyapatite cement influence the resorption and bony substitution has to be shown in further studies.

Animals↗
PubMed1991

The effects of human salivary cystatins and statherin on hydroxyapatite crystallization.

The adsorption, at hydroxyapatite surfaces of neutral cystatin SN, acidic cystatin S and the phosphoserine-containing acidic cystatin S1 was compared to that of statherin. The effects of these adsorbed proteins on the constant-composition growth kinetics of hydroxyapatite were also studied. The neutral cystatin SN had a higher adsorption maximum than the acidic cystatins S and S1. Although the affinity of cystatin for hydroxyapatite surfaces was lower than that of statherin, their influence on the growth kinetics of hydroxyapatite was considerably greater, with the acidic cystatin S1 being the most active. At a surface concentration of 7.0 x 10(-8) mol m-2 hydroxyapatite, the cystatins decreased the rate of crystal growth by 80-95% as compared to that in the absence of protein. At this concentration, statherin showed a growth inhibition of 40%.

Adsorption↗
PubMed1993

The effect of crystallinity on hydroxyapatite-induced production of reactive oxygen metabolites by polymorphonuclear leukocytes.

To assess the role of crystallinity in biological response, we quantitated the generation of reactive oxygen metabolites in human polymorphonuclear leukocytes (PMN) with a chemiluminescence assay using three hydroxyapatite preparations with sintering temperatures of 1,200 degrees C and 900 degrees C and a drying temperature of 110 degrees C on the basis of equal weights (1 mg/ml). These crystals have almost the same average diameters and similar average zeta potentials. The crystals prepared at higher temperatures have higher crystallinity, or larger domain sizes, which were calculated by X-ray diffraction line broadening. The production of reactive oxygen metabolites by PMN in hydroxyapatite of 1,200 degrees C was 10-times that by PMN in hydroxyapatite of 900 degrees C and more than 50-times greater than that in hydroxyapatite of 110 degrees C. A single linear correlation was observed for a plot of log (peak chemiluminescence levels) vs. a plot of log (domain sizes). These results clearly show that the maximal effect of crystallinity on hydroxyapatite-induced production of reactive oxygen metabolites by human PMN was seen at higher crystallinity.

Crystallization↗
PubMed1987

A histologic evaluation of a mandibular cross section one year after augmentation with hydroxyapatite particles.

A pathologic mandibular fracture in a 35-year-old woman 1 year after the attempted filling of a bone defect with particulate, dense hydroxyapatite resulted in a partial mandibular resection, allowing observation of cross sections of the mandible and associated soft tissue. Thin tissue sections were examined without decalcification. This permitted excellent observation of the hydroxyapatite particles, the bone, and the soft tissue. Osseointegration had occurred only in the areas closely associated with the bone. The immobility of the particles was a prerequisite for involvement with new bone formation. Hydroxyapatite particles in areas that allowed any mobility were surrounded by connective tissue with no bone formation evident. Heterogeneous particles were observed, indicating the possibility of a lack of purity in the hydroxyapatite ceramic, which may have contributed to the resorption of the particles. These findings, as well as other clinical and experimental findings, lead us to question the concept of hydroxyapatite as a bioactive ceramic that induces osteogenesis or osteoconductivity.

Adult↗
PubMed1992

Heterogeneous synthesis of fluoridated hydroxyapatites.

Fluoridated hydroxyapatites were synthesized with two different modes of fluoride supply: by supplying F(-)-free solution initially, followed by a F(-)-containing solution whose fluoride concentration was stoichiometrically equal to that of fluorapatite; and with the order of supply of these solutions reversed. Both of these heterogeneously synthesized fluoridated hydroxyapatites showed typical calcium phosphate X-ray diffraction patterns; and both had similar total fluoride contents (0.94 +/- 0.02 and 0.99 +/- 0.03 mmol/g, respectively), i.e. half of the maximum fluoride content of fluorapatite. However, they differed considerably in their physico-chemical properties. The former had a wider peak breadth of X-ray diffraction and a lower apparent solubility (Ca = 12.2 +/- 0.7 mmol/l) than the latter (Ca = 19.8 +/- 0.4 mmol/l) at 37 degrees C and pH 4.0. Wavelength dispersive spectroscopy attached to scanning electron microscopy gave clearly different spectra. Electron spectroscopy for chemical analysis of apatite pellets made by pressing and heating showed a slightly larger F1s signal at the crystal surface of the former, although the difference was not marked. These results suggest that two different types of heterogeneous fluoridated hydroxyapatites were formed, hydroxyapatite covered with fluorapatite and fluorapatite covered with hydroxyapatite.

Fluorides↗
PubMed1992

Interface between hydroxyapatite and mandibular human bone tissue.

Samples from intraosseous dental implants, removed from patients for mechanical failures, were examined to analyse the interaction between hydroxyapatite as plasma sprayed coating on titanium supports and human bone. The implantation time varied up to 8 years. No failures had arisen from problems at the interface between the hydroxyapatite coating and bone. The number of samples examined and the implantation times give good statistical conclusions. Histological and microchemical studies showed the good performance and compatibility of this sprayed hydroxyapatite. We present evidence from the best samples which show close bonding with the surrounding bone tissue. New bone is seen all around the coated implant. The composition of the calcium phosphate deposited on the hydroxyapatite and cellular approach were determined, and demonstrate the efficiency of the interaction between this plasma sprayed hydroxyapatite and the bone.

Adhesiveness↗
PubMed1993

Amorphous phase and morphological structure of hydroxyapatite plasma coatings.

It is shown that under standard conditions of plasma spraying hydroxyapatite powder reaches the substrate in the form of molten drops. On striking the surface, the molten drop splatters, cools rapidly and solidifies. Consequently, a pancake particle surrounded by tiny spherical or differently shaped particles is formed on the substrate. The solidified particles are partly crystallized. Besides the crystalline hydroxyapatite, they contain some supercooled molten hydroxyapatite, i.e. amorphous hydroxyapatite. Build-up of these particles during coating gives a porous pancake-globular structure. During a brief vacuum heating at 630 degrees C, the amorphous phase is crystallized and completely crystalline hydroxyapatite coatings are obtained.

Crystallization↗
PubMed1996

Development of hydroxyapatite derived from Indian coral.

A simple method of converting the calcium carbonate skeleton of the corals available in the Indian coast into hydroxyapatite granules has been developed. By heating the coral to 900 degrees C, the organic materials were eliminated. Powder X-ray diffraction analysis (XRD), Fourier transform infrared spectroscopy (FTIR) and thermogravimetric analysis (TGA) were employed to characterize the coral and to optimize the processing parameters as well as to confirm the hydroxyapatite formation. The coral used exhibits the presence of both aragonite and calcite phases (dimorphism). At a temperature of 900 degrees C the coral was found to decompose all the carbonate phases. The pre-heated coral is converted into hydroxyapatite by a chemical exchange reaction with di-ammonium phosphate under hydrothermal conditions. The hydroxyapatite obtained was in powder form and does not contain any impurities. The in vitro solubility test of the apatite granules performed in Gomoris, Michalelis, Sorensens, Ringer's and phosphate buffer of pH 7.2 and de-ionized water indicated the stability of the coralline hydroxyapatite.

Animals↗
PubMed1991

Preferential adsorption of dentin and bone acidic proteins on the (100) face of hydroxyapatite crystals.

Interaction of bone and dentin proteins with minerals is an elementary step in the regulation of mineralization in these tissues. Adsorption of acidic non-collagenous proteins on hydroxyapatite was examined using fluorescence-labeled protein and synthetic hydroxyapatite. Phosphophoryn, bone Gla protein, osteonectin and bone small proteoglycan II were prepared and labeled with fluorescein. All of these proteins were adsorbed on hydroxyapatite with a dissociation constant on the order of 10(-7) M. The more acidic proteins had lesser binding capacities. Hydroxyapatite single crystals were incubated with labeled proteins and observed with a fluorescence microscope. Phosphophoryn and other acidic proteins were adsorbed preferentially on the (100) face of the crystal. This preferential adsorption of the acidic proteins may be responsible for the morphogenesis of biological hydroxyapatite.

Animals↗
PubMed2000

Discoloration of ceramic hydroxyapatite used for protein chromatography.

Hydroxyapatite chromatography was used to purify a recombinant human protein at preparative (400 g) scale. The hydroxyapatite column became progressively discolored as the number of chromatographic cycles increased. Elemental analysis showed that Mn, Fe, Al, Cd, Ba, Cr and Sn were found in used, discolored hydroxyapatite but were below the detection limit (1 ppm) in new hydroxyapatite. Metal ions were not removed from the discolored hydroxyapatite by regeneration with 0.5 M sodium phosphate followed by 0.5 M sodium hydroxide. Chromatographic performance was not affected by the accumulation of metal ions for at least 8 cycles on the preparative column (media volume 56 l) and for at least 12 cycles on the laboratory-scale column (media volume 3.7 ml).

Ceramics↗
PubMed1989

Eleven-year study of hydroxyapatite implants.

An 11-year clinical research study was conducted with both unloaded bulk hydroxyapatite implants and loaded hydroxyapatite-coated titanium implants. A total of 102 submerged bulk hydroxyapatite implants were placed after extraction of teeth to maintain the volume of the residual alveolar ridge by their physical presence. All 21 implants under fixed partial dentures and 51 of 81 implants under lower complete dentures remained submucosal. A total of 71 hydroxyapatite-coated titanium implants were connected with permucosal superstructures by use of a two-stage method. Modifications in design and in implantation technique were required. This long-term research indicates that cylindrical hydroxyapatite implants are reliable devices as natural tooth root substitutes that bond directly to bone instead of simply being osseointegrated.

Dental Abutments↗
PubMed1998

Critical ageing of hydroxyapatite sol-gel solutions.

It has been established that hydroxyapatite coatings can be produced using an alkoxide based sol-gel technique. Previous work showed that in addition to hydroxyapatite other phases including CaO were observed. A critical factor in determining the composition of the final coating is the period between solution preparation and coating deposition. The current study addresses how the ageing time affects the composition of the coating and the time required before a solution can be used to deposit monophasic coatings. Powders and coatings were produced with ageing times up to one week and examined with X-ray diffraction, thermal gravimetric analysis and differential thermal analysis. By depositing coatings after various ageing times, it was found that a solution ageing time of at least 1 day was required before monophasic hydroxyapatite coatings could be deposited. Thermogravimetric analysis showed that heating of the gel in air or in nitrogen both produced hydroxyapatite. Thermogravimetric analysis could be used as a simple and effective tool for identifying the critical ageing time required to produce high-purity hydroxyapatite coatings.

Biocompatible Materials↗