Solid state 31NMR studies of the conversion of amorphous tricalcium phosphate to apatitic tricalcium phosphate.

The hydrolytic conversion of a solid amorphous calcium phosphate of empirical formula Ca9 (PO4)6 to a poorly crystalline apatitic phase, under conditions where Ca2+ and PO4(3-) were conserved, was studied by means of solid-state magic-angle sample spinning 31P-NMR (nuclear magnetic resonance). Results showed a gradual decrease in hydrated amorphous calcium phosphate and the formation of two new PO4(3-)-containing components: an apatitic component similar to poorly crystalline hydroxyapatite and a protonated PO4(3-), probably HPO4(2-) in a dicalcium phosphate dihydrate (DCPD) brushite-like configuration. This latter component resembles the brushite-like HPO4(2-) component previously observed by 31P-NMR in apatitic calcium phosphates of biological origin. Results were consistent with previous studies by Heughebaert and Montel [18] of the kinetics of the conversion of amorphous calcium phosphate to hydroxyapatite under the same conditions.

Calcium phosphate ceramic coatings on porous titanium: effect of structure and composition on electrophoretic deposition, vacuum sintering and in vitro dissolution.

Bioactive calcium phosphate ceramics (CPC) guide bone formation along their surface. This property is conceptually attractive from the viewpoint of enhancing early bone tissue formation in porous metal coatings. The various studies conducted to exploit this idea, however, reveal a considerable variability of the effect. This suggests material- and processing-induced parametric influences. Thus this study focuses on the formulation of model porous metal-CPC materials for use in one-parametric analyses of material factors. Easily reproducible, porous metals with a uniform porous structure and CPC coating are made with orderly oriented wire mesh (OOWM) porous metal coatings and electrophoretically deposited CPC films. The deposition of the ceramic can be hampered by adsorbed water. Subsequent vacuum sintering leads to several phase transformations: hydroxyapatite is transformed to a mixture of oxyhydroxyapatite and tetracalcium phosphate; the underlying titanium promotes the beta- to alpha-tricalcium phosphate transformation; and Ca-deficient hydroxyapatite is transformed to a mixture containing oxyhydroxyapatite and alpha- and beta-tricalcium phosphate. These phase transformations provoke a considerable increase of in vitro dissolution in 0.05 M tris buffered physiological solution.

Formation of carbonate-apatite crystals after implantation of calcium phosphate ceramics.

The aims of this study were (1) to determine at the crystal level, the nonspecific biological fate of different types of calcium phosphate (Ca-P) ceramics after implantation in various sites (osseous and nonosseous) in animals and (2) to investigate the crystallographic association of newly formed apatitic crystals with the Ca-P ceramics. Noncommercial Ca-P ceramics identified by X-ray diffraction as calcium hydroxylapatite (HA), beta-tricalcium phosphate (beta-TCP), and biphasic calcium phosphates (BCP) (consisting of beta-TCP/HA = 40/60) were implanted under the skin in connective tissue, in femoral lamellar cortical bone, articular spine bone, and cortical mandibular and mastoidal bones of animals (mice, rabbits, beagle dogs) for 3 weeks to 11 months. In humans, HA or beta-TCP granules were used to fill periodontal pockets, and biopsies of the implanted materials were recovered after 2 and 12 months.(ABSTRACT TRUNCATED AT 250 WORDS)

Chemical changes in hydroxyapatite biomaterial under in vivo and in vitro biological conditions.

The introduction of a synthetic calcium phosphate into a biological environment is likely to result in surface-mediated chemical events. On the basis of such an assessment, we studied the chemical changes occurring in the mineral after exposure of a synthetic hydroxyapatite ceramic to both in vivo (implantation in human) and in vitro (cell culture) conditions. A small amount of the material was phagocytized but the major remaining part behaved as a secondary nucleator as evidenced by the appearance of a newly formed mineral. Morphologically, the newly formed mineral appeared as tiny crystals precipitated and grown from the surface of the initial synthetic crystals. The density of the additional mineral increased from the periphery to the core of each biomaterial aggregate. Chemically, it was identified by IR spectroscopy as a carbonated apatitic mineral. We propose that the adsorption of biomolecules could inhibit precipitation, accounting for the increasing amount of precipitate from the periphery to the core of the aggregates.

Hydroxyapatite biomaterial implanted in human periodontal defects: an histological and ultrastructural study.

The purpose of the present work was to study the response of human periodontium to hydroxyapatite biomaterial particles (180-200 microns). The biomaterial was implanted in two infra-osseous periodontal defects (two patients) after clearing of the granulation tissue. At two months post-surgery, biopsies were studied using light and electron microscopy. No sign of inflammation was observed, the biomaterial aggregates were surrounded either by typical fibroblasts or larger phagocytotic cells with phagocytosis vesicles containing biomaterial crystals. These intracellular crystals were noticeably smaller than the non-phagocytized ones. Some of the phagocytized crystals showed morphological signs of intracellular dissolution. The spaces between the crystals constitutive of the aggregates were filled with organic substance containing collagen fibers.

Physicochemical characterization of deposits associated with HA ceramics implanted in nonosseous sites.

Pellets of well-characterized microporous hydroxyapatite (HA) ceramic were implanted in hamsters in two nonosseous sites: (1) in the fatty tissue of the gingival crease, far from bony tissue and (2) in intraperitoneal sites. The implants in site 1 were placed directly in contact with tissues, cells, and extracellular fluids while the implants in site 2 were placed in special chambers made of plexiglass cylinders covered in both ends with millipore filters, preventing contact with tissues and cells, but not with extracellular fluids. The hamsters were sacrificed and the implants recovered after 8, 16, 30, 150, and 365 days. The pellets were characterized using x-ray diffraction, infrared absorption, thermogravimetry, scanning and transmission electron microscopy, and calcium and phosphate analyses before and after implantation. Physicochemical analyses of HA ceramic implants before and after implantation demonstrated the formation of new material which was significantly different from the HA ceramic in terms of the following: (a) morphology (size of shape) of crystals; (b) intimate association of the inorganic phase of the new material with an organic phase similar to inorganic/organic association in bone; (c) the inorganic phase of the new material is a CO3-apatite, similar to that of bone, while the HA in ceramic is CO3-free; (d) electron diffraction of apatite of new material is similar to that of bone apatite. This study also demonstrated that the new material associated with the HA ceramics implanted in two different nonosseous sites were identical in spite of the differences in their microenvironment (cellular and acellular).

In vivo cell interactions with calcium phosphate bioceramics.

Biointegration, resorption process, and solubility in physiological environments of calcium phosphate materials are scarcely described by ultrastructural studies. In vivo cells interactions with calcium phosphate materials are scarcely described by ultrastructural studies. In vivo cells interactions with calcium phosphate biomaterials are mediated by different proteins from physiological fluid, and in order to observe at the ultrastructural level the cell colonization, the resorption, process and the biointegration, we used in these experiments calcium phosphate materials precoated with fibronectin or not precoated. Two kinds of well determined materials were used for this study, Beta-tricalcium phosphate (B-TCP) and hydroxyapatite (HAP). The implants were soaked in human fibronectin diluted solution and were implanted in the connective tissue of rabbit abdomen. Our results showed that the fibroblasts and macrophagous++ cells interaction with the calcium phosphate crystal (B-TCP and HAP) was more important in the experiments with a fibronectin bilayer. In the presence of fibronectin at the grains surface of the material, cystic cavities' or fibrous encapsulation was suppressed and cells with fibers were in close contact with the material. The presence of fibronectin immediately after implantation seemed to increase the adhesion and the cell colonization. Fibronectin creates an organic interface between crystals and cells, and can promote interactions from cells and biomaterials.

Apatitic calcium orthophosphates and related compounds for biomaterials preparation.

The authors show that to obtain well chemically defined apatitic bioceramics and to know the possible transformations of this material during sintering, it is necessary to prepare a good starting material. Moreover, they show that it is possible to prepare a new organic-inorganic phosphate compound. The precipitation of apatite in an aqueous medium at boiling temperature was studied using the methodology of experimental design. Independent variables were the volume of NH4OH in phosphate solution, the volume of NH4OH in calcium solution, and the time of precipitation; the response was the atomic Ca/P ratio of the obtained precipitate. A continuous variation of this ratio from 1.63 to 1.73 is observed. Implications of this result to the preparation of pure HA: Ca10(PO4)6(OH)2 is given. Moreover, when Ca/P greater than 1.67, HA reacts with Ca(OH)2 (after heating at 1000 degrees C in air for some days) to give rise to a single phase described as a modified HA (MHA), a Ca/P ratio of 1.75, an a value of 9.373 +/- 0.002 A, and a c value of 6.884 +/- 0.002 A. The reactivity (time versus temperature) of the MHA is described. If the precipitation of the calcium phosphate is realized at 37 degrees C in a water-ethanol medium in the presence of A2EP, a new apatite, chemically bonded to the organic molecule by pooling phosphate groups, is obtained.

[Ultrastructural study of bone formation after bioapatite implantation in man]. / Etude ultrastructurale de l'induction osseuse après implantation de bioapatites chez l'homme.

Synthetic hydroxyapatite crystals (Bioapatite) were implanted in a vestibular longitudinal groove made in the alveolar bone, the periodontal membrane and the superficial root dentine of 2 human canines in the absence of periodontal disease. The healing process was followed in scanning as well as in transmission electron microscopy. At 6 months, the aggregate of synthetic hydroxyapatite crystals was surrounded by a fibrous connective tissue, devoid of inflammatory cells. Small apatite crystals, similar to the neighbouring bone crystals, were deposited between the Bioapatite crystals in the central area of the aggregates. These small crystals filled the intercrystalline spaces from the center to the periphery. The mineralization extended then directly in the collagen matrix (osteoid tissue) surrounding the synthetic apatite aggregates. Bone tissue was thus formed around the apatite implants. At 1 year, the thickness of the bone tissue formed around the synthetic hydroxyapatite aggregates was increased. It consisted of bone with a well-oriented collagen matrix, with osteocyte lacunae, osteocytes and interosteocytic canaliculi, with normal ultrastructure.

Structural analysis of hydroxyapatite coatings on titanium.

Hydroxyapatite from two sources was electrophoretically deposited onto flat titanium plate material. Depending upon the deposition conditions various changes in the structure of the ceramic were identified. A well-adhering Ti-P compound was present at the interface. Hydroxyapatite oxygenated to various degrees and tetracalcium phosphate were reproducibly formed in the coating.

[Histological study of the biocompatibility of hydroxyapatite crystals in periodontal surgery]. / Etude histologique de la biocompatibilité des cristaux d'hydroxyapatite en chirurgie parodontale.

The purpose of this investigation was to analyse, after 6 years of clinical trials, the findings obtained from biopsies of infrabony defects in man treated by grafts of hydroxyapatite. Filled extraction sockets in miniature swine treated in a similar manner with apatite were also studied. Light microscope analysis showed bone neoformation around and in the neighbourhood of grafted apatite fragments with the presence of osteocytes, osteoblasts and a normal peripheral connective tissue without inflammatory reaction. These results demonstrate the biocompatibility of the implanted apatites accompanied by a normal fibrogenesis and a seeming osteogenesis.