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Processing of an apatite-mullite glass-ceramic and an hydroxyapatite/phosphate glass composite by selective laser sintering.

The work presented details the results of an investigation into the feasibility of using Selective Laser Sintering (SLS) to directly produce customised bioceramic implants. The materials used were bioactive in nature and included a glass-ceramic and a combination of hydroxyapatite and phosphate glass. The glass-ceramic was selected from the range of apatite-mullite materials in the SiO2.Al2O3.CaO.CaF2.P2O5 series, due to their potentially suitable biological and mechanical properties. The hydroxyapatite and phosphate glass combination was chosen to allow an alternative production approach to be investigated. The viability of using both these materials with the SLS process was assessed and the process route and resulting material properties characterised using a variety of techniques including Differential Thermal Analysis (DTA), X-ray Diffraction (XRD) and Scanning Electron Microscopy (SEM). The results obtained indicate that it was possible to produce multiple layer components from both materials using the SLS process. The glass-ceramic materials could only be processed at very low scan speeds and powers, yielding relatively brittle components. It was though possible to produce parts from the hydroxyapatite and phosphate glass combination across a much wider range of parameters, producing parts which had a greater potential for possible implant production.

Aluminum Silicates↗

Apatite formation on poly(2-hydroxyethyl methacrylate)-silica hybrids prepared by sol-gel process.

Hybrids of poly(2-hydroxyethyl methacrylate) (PHEMA), a polymer that has been employed in a wide variety of biomedical applications, and silica-gel, which exhibits a well-known bioactivity, were produced. The obtained hybrids were characterized and their in vitro ability to induce the formation of a calcium phosphate layer on the surface was evaluated. The surface area of hybrids decreased with increasing amounts of PHEMA so that hybrids with more than approximately 40% PHEMA are virtually non-porous. All hybrids induced the formation of a calcium phosphate layer on their surfaces when soaked into simulated body fluid. The induction time and the morphology of the apatite layer varied according to the polymer content.

Apatites↗

In vitro apatite forming ability of type I collagen hydrogels containing bioactive glass and silica sol-gel particles.

Type I collagen hydrogel containing bioactive glass (CaO-P2O5-SiO2) and silica sol-gel micrometric particles were prepared and their in vitroapatite-forming ability in simulated body fluid assessed. X-ray diffraction and scanning electron microscopy analysis showed that bioactive glass particles entrapment in collagen matrix did not inhibit calcium phosphate formation and induced morphology variations on the crystalline phase precipitated on the hydrogel surface. The silica--collagen hydrogel composite precipitated calcium phosphate whereas silica particles and collagen hydrogel alone did not, indicating a possible synergetic effect between collagen and silica on the apatite-forming ability. Mechanisms of calcium phosphate precipitation and its relevance to biomaterial development are discussed.

Apatites↗

Clinical evaluation of castable apatite ceramic crowns.

Thirty-three castable apatite ceramic crowns were constructed for 26 patients. The crowns were clinically evaluated for anatomic form, marginal adaptation, cavosurface marginal discoloration, surface roughness, and color match for a period of 2 years at regular intervals. Results showed that all restoration had satisfactory anatomic form; all but one exhibited good marginal adaptation. Slight abrasion was noticed at the functioning cusps and all but three of the restorations demonstrated excellent color matching.

Adult↗

Compositional variations in apatites with respect to preferential ionic extraction.

Detection of ionic losses from the apatitic structure (Ca10(PO4)6(OH)2) by high-resolution electron microscopy was investigated theoretically. Linear image analysis showed the need for an objective aperture of at least 3.7 nm-1 to visualize four different coordinates (CaII, OH-, P and mid-point between CaII-P bond). High-resolution image analysis and plotting of OH- column intensity against specimen thickness showed an inverse proportionality between composition and OH- image intensities for very thin specimens (less than 2 nm). Image intensity variation would be detectable experimentally, but the preparation of such thin specimens by ultramicrotomy is impossible.

Apatites↗

Chemical and biological integration of a mouldable bioactive ceramic material capable of forming apatite in vivo in teeth.

Chemically bonded ceramics have several advantages compared with conventional ceramics to be used as biomaterials. Especially the possibilities to harden the material at room temperature and to control the rheology are very beneficial. This paper investigates the interface formed in vivo between a calcium aluminate based dental filling material and teeth. Class 1 occlusal fillings were made in wisdom teeth and extracted after up to four weeks. Polished cross-sections of the teeth were studied with scanning electron microscopy (SEM), focused ion beam microscopy (FIB) and transmission electron microscopy (TEM). In order to analyse the distribution of elements at the interface elemental mapping was performed using STEM and EDX. The results showed that a tight bond forms between the filling material and tooth and no gap could be found even at high magnification. A 100-200 nm wide zone with an increase in oxygen was detected in the enamel next to the filling. The zone was denser than the rest of the enamel. Elemental mapping indicated an increase of silicon and a decrease of Ca at the interface. Dark field imaging and EDX mapping showed that the calcium aluminate system formed apatite in situ during hardening through precipitation.

Aluminum Compounds↗

Proliferation and differentiation of osteoblast-like cells on apatite-wollastonite/polyethylene composites.

Glass-ceramic apatite-wollastonite (A-W)/high-density polyethylene composite (AWPEX) materials have been designed to match the mechanical strength of human cortical bone and to provide favourable bioactivity, with potential use in many orthopaedic applications. To better understand AWPEX properties, the effects of surface finish and ceramic filler size and content on osteoblast-like cell attachment, proliferation, and differentiation were examined. Glass-ceramic content was tested at 30 and 50 vol% and median particle size at 4.5 and 7.7 microm. Samples were prepared as 1 x 10 x 10 mm(3) tiles with polished or rough surfaces, sterilized by gamma irradiation (2.5 Mrad), and characterized by scanning electron microscopy (SEM) and surface profilometry. Saos-2 human osteoblast-like cells were cultured on each surface at an initial concentration of 4500 cells/cm(2) for 1, 3, or 7 days. At each time point, adenosine triphosphate and alkaline phosphatase levels were measured to assess cell number and osteoblast differentiation. SEM imaging of cells on the composite surfaces showed preferential cell attachment to filler particles within the polymer matrix. Significant biochemical assay differences were found at 7 days, confirmed by ANOVA post-hoc testing using Bonferroni's correction. Overall, increased exposure of the glass-ceramic A-W phase in AWPEX through surface polishing, higher volume fraction and/or larger particle size was found to lead to an improved cell response.

Apatites↗

Electron spin resonance in silicon substituted apatite and tricalcium phosphate.

Impurity centers associated with silicon have been observed in the phase mixture of silicon substituted apatite (Si-Ap) and silicon stabilized tricalcium phosphate (Si-TCP) using electron spin resonance (ESR). Two unique centers occur upon addition of SiO2 to the calcium phosphate system: an orthorhombic center with g-values 2.0072+/-0.0001, 2.0024+/-0.0001 and 2.0003+/-0.0001 (Si-h1) and a center with tetrahedral symmetry having g-values components 2.0054+/-0.0001 and 1.9992+/-0.0003 (Si-h2). Both centers are hypothesized to be characteristic of defects associated with silicon in the Si-Ap phase. Through comparison of the intensity of F-OH centers in undoped calcium hydroxyapatite (HA) prepared with various levels of OH occupancy, a relationship is demonstrated between the ESR intensity of an F-center signal with g = 2.0019+0.0004 (F-OH) and the OH occupation of HA. Relative changes in the intensity of ESR signals Si-h1 and F-OH are consistent with a chemical model describing the substitution of SiO4(4-) for PO4(3-) in HA with the creation of OH- vacancies as charge compensation, resulting in a mixed phase composition of Si-Ap and Si-TCP that results when a hydroxyapatite precipitate (HA) is heated in the presence of added SiO2.

Apatites↗

Bone tissue engineering on amorphous carbonated apatite and crystalline octacalcium phosphate-coated titanium discs.

Poor fixation of bone replacement implants, e.g. the artificial hip, in implantation sites with inferior bone quality and quantity may be overcome by the use of implants coated with a cultured living bone equivalent. In this study, we tested, respectively, amorphous carbonated apatite (CA)- and crystalline octacalcium phosphate (OCP)-coated discs for their use in bone tissue engineering. Subcultured rat bone marrow cells were seeded on the substrates and after 7 days of culture, the implants were subcutaneously implanted in nude mice for 4 weeks. After 7 days of culture, the cells had formed a continuous multi-layer that covered the entire surface of the substrates. The amount of cells was visually higher on the crystalline OCP-coated discs compared to the amorphous CA-coated discs. Furthermore, the amorphous CA-coated discs exhibited a visually higher amount of mineralized extracellular matrix compared to the crystalline OCP-coated discs. After 4 weeks of implantation, clear de novo bone formation was observed on all discs with cultured cells. The newly formed bone on the crystalline OCP-coated discs was more organized and revealed a significantly higher volume compared to the amorphous CA-coated discs. The percentage of bone contact with the discs was also significantly higher on the OCP-coated discs. Overall, the results suggest that a crystalline OCP coating is more suitable for bone tissue engineering than an amorphous CA coating.

Animals↗

The role of cell signaling defects on the proliferation of osteoblasts on the calcium phosphate apatite thin film.

The intracellular signal transduction controlling the proliferation of osteoblastic cells on a thin film of poorly crystalline calcium phosphate apatite crystals (PCA) was studied in vitro. The PCA thin film was prepared on polystyrene culture dishes or cover glasses using phosphate-buffered calcium ion solution, which was made oversaturated by heating an undersaturated solution prepared at low temperature. The PCA thin film was used for cell culture without additional surface treatment. Several differences were found between the cells plated on a cell culture dish and the cells cultured on the PCA surface. Entry into S-phase of the cell cycle was markedly delayed and there was a low proliferation rate of osteoblast. On the PCA thin film, the cells spread in a more slender shape. Also, the formation of focal adhesions and stress fibers, examined using immunocytochemical staining, was strikingly weaker in the cells on PCA. The activation of focal adhesion kinase (FAK) was low as well. Expression of cyclins D1 and E was also lower. The Ras-Extracellular signal-regulated kinase (ERK)-MAP kinase signaling pathway was weakly activated by stimulation with serum. These results demonstrate that the low cell proliferation on the PCA surface appears to be due to insufficient activation of signaling that forces the cell cycle to progress and this may be due to weak adhesion signaling in cells on that surface.

3T3 Cells↗

A comparative study of the metastable equilibrium solubility behavior of high-crystallinity and low-crystallinity carbonated apatites using pH and solution strontium as independent variables.

UNLABELLED: Using solution strontium and pH as independent variables, the metastable equilibrium solubility (MES) behavior of two carbonated apatite (CAP) samples has been examined, a high-crystallinity CAP (properties expected to be similar to dental enamel) and a low-crystallinity CAP (properties expected to be similar to bone mineral). CAP samples were prepared by precipitation/digestion: (CAP A: high-crystallinity, 1.3 wt% CO3, synthesized at 85 degrees C; CAP B: low-crystallinity, 6.4 wt% CO3, synthesized at 50 degrees C). Baseline MES distributions were determined in a series of 0.1 M acetate buffers containing only calcium and phosphate (no strontium) over a broad range of solution conditions. To assess the influence of strontium, MES profiles were determined in a similar fashion with 20, 40, 60, and 80% of the solution calcium being replaced on an equal molar basis by solution strontium. To determine the correct function governing CAP dissolution, ion activity products (IAPs) were calculated from the compositions of buffer solutions based on the hydroxyapatite template (Ca(10-n)Sr(n)(PO4)6(OH)2 (n = 0-10)) and the calcium/hydroxide deficient hydroxyapatite template (Ca(9-n)Sr(n)(HPO4)(PO4)5OH (n = 0-9)). FINDINGS: (a) for CAP A, at high solution strontium/calcium ratios, the MES profiles were essentially superimposable when the solution IAPs were calculated using the stoichiometry of Ca6Sr4(PO4)6(OH)2 and for CAP B by a stoichiometry of Ca7Sr2(HPO4)(PO4)5OH; (b) for CAP A, at low strontium/calcium ratios, the stoichiometry yielding MES data superpositioning was found to be that of hydroxyapatite and for CAP B, that of calcium/hydroxide deficient hydroxyapatite. When other stoichiometries were assumed, good superpositioning of the data was not possible.

Apatites↗

Efficiency of 1H --> 31P NMR cross-polarization in bone apatite and its mineral standards.

Human bone mineral was studied using solid-state 31P NMR with cross-polarization (CP) from protons. The CP efficiency was determined for trabecular and cortical bone tissue from human adults and compared with synthetic mineral standards. The study shows the similarity between carbonatoapatite of type B and bone mineral as shown by their CP behaviour. The method can be used for the characterization of synthetic apatite-based implant materials.

Adult↗

Theoretical detection of a dark contrast line in twinned apatite bicrystals and its possible correlation with the chemical properties of human dentin and enamel crystals.

Electron microscope images of twinned apatite bicrystals oriented along the [1120] crystallographic direction have been simulated for various experimental conditions, and the validity of the calculation has been checked. These images show a dark contrast line similar to the one observed experimentally in enamel and dentin crystals and therefore strongly suggest the presence of a twin plane parallel to the (1100) crystallographic planes, in these crystals. The presence of a twin boundary in teeth and bone crystals is of prime importance for the adsorption and the dissolution properties of the calcified tissues as a whole.

Apatites↗

Influence of zeolite, apatite and Fe-oxide on Cd and Pb uptake by crops.

Natural zeolite (clinoptilolite), hydroxyapatite and an iron-oxide waste by-product (Fe-rich, a trademark name of E.I. du Pont de Nemours) were added to an artificially contaminated Appling soil to immobilize and limit the uptake of metals by crops. A greenhouse pot study employed spiking the soil with Cd and Pb from metal flue dust. Maize (Zea mays) and barley (Hordeum vulgare) were planted in 7-kg potted soil to determine the effects of Cd and Pb on plant growth and uptake. Sequential extraction of soil indicates the substantial influence of soil pH and type of ameliorant on the chemical form and bioavailability of the metals. Data indicates that a dose of 50 g/kg of soil of iron-oxide appears to be very effective, based on the yields, metal contents of plant tissues and available forms of Cd and Pb in the soil. Lower doses of zeolite and apatite (15 g/kg and 4 g/kg soil, respectively) in most cases also reduced significantly the uptake of Cd and Pb by crops.

Apatites↗

Fabrication of Zn containing apatite cement and its initial evaluation using human osteoblastic cells.

Recently, the effects of Zn2+ on osteogenesis stimulation have become major topics in the research fields of bone formation and organism essential elements. Based on the fundamental finding of Zn2+ with respect to osteogenesis stimulation, Ito et al. have prepared Zn doped beta-tricalcium phosphate (ZnTCP) and have reported that ZnTCP enhances the proliferation of MC3T3-E1 cells. In this investigation, we studied the effects of ZnTCP added to apatite cement (AC) with respect to its setting reaction and proliferation of human osteoblastic cells as an initial evaluation for the feasibility of AC containing ZnTCP. Compositional analysis using powder X-ray diffractometer revealed that ZnTCP shows no reactivity with the setting reaction of AC. As a result, the mechanical strength of set AC decreased increasing amounts of added ZnTCP as if ZnTCP acts as a pore in AC. The setting time of AC was not affected by addition of ZnTCP up to 10%. When AC containing ZnTCP was immersed in alpha-MEM containing 10% bovine serum, Zn2+ was released from AC. Larger amounts of Zn2+ were released from AC containing larger amounts of ZnTCP. When human osteoblastic cells were incubated on the surface of AC discs, proliferation of human osteoblastic cells was significantly increased on the surface of AC that contained 5% ZnTCP when compared with that containing no ZnTCP. In contrast, proliferation of human osteoblastic cells decreased on the surface of AC that contained 10% ZnTCP when compared with that free from ZnTCP; indicating cytotoxicity. We concluded therefore, that addition of ZnTCP to AC is useful to enhance the osteoconductivity of AC when release of Zn2+ can be carefully regulated.

Apatites↗

Mechanochemical-hydrothermal synthesis of carbonated apatite powders at room temperature.

Crystalline carbonate- and sodium-and-carbonate-substituted hydroxyapatite (CO3HAp and NaCO3HAp) powders were prepared at room temperature via a heterogeneous reaction between Ca(OH)2/CaCO3/Na2CO3 and (NH4)2HPO4 aqueous solution using the mechanochemical hydrothermal route. X-ray diffraction, infrared spectroscopy, thermogravimetry, and chemical analysis were performed. Room temperature products were phase-pure CO3HAp and NaCO3HAp containing 0.8-12 wt% of carbonate ions in the lattice. Dynamic light scattering revealed that the median agglomerate size of the room temperature CO3HAp and NaCO3HAp powders was in the range of 0.35-1.6 microm with a specific surface area between 82 and 121 m2/g. Scanning and transmission electron microscopy confirmed that the carbonated HAp powders consisted of mostly submicron aggregates of nanosized, approximately 20 nm crystals. The synthesized carbonated apatite powders exhibit chemical compositions and crystallinities similar to those of mineral constituents of hard tissues and therefore are promising for fabrication of bone-resembling implants.

Apatites↗

Initial tissue response to a titanium implant coated with apatite at room temperature using a blast coating method.

We previously reported a blast coating method (BC method) as a new coating method of titanium (Ti) with apatite (AP) at room temperature. The BC method gives much stronger AP coating on the Ti surface compared with those obtained by other room temperature coating methods. However, no in vivo study has been made, so far, to evaluate the stability or the tissue response to the implant. As an initial step to evaluate the feasibility of the BC method, we evaluated the tissue response and stability of AP coated Ti implant prepared with the BC method (AP-BC implant) using rats as experimental animals. The AP coating adhered tightly to the Ti surface even after the implant procedure and throughout the experimental period up to six weeks post operation. AP-BC implant caused no inflammatory response, showed strong bone response and much better osteoconductivity compared with the pure Ti implant. The new bone formed on the surface of AP-BC implants was thinner compared with that formed on the surface of Ti implant. Therefore, the AP-BC implant has a good potential as an osteoconductive implant material.

Animals↗

Transformation mechanism of different chemically precipitated apatitic precursors into beta-tricalcium phosphate upon calcination.

The Ca-deficient apatite (CDHA) was prepared from the precursors of (CH3COO)2Ca x xH2O, Ca(NO3)2 x 4H2O and H3PO4, (NH4)H2PO4 to investigate the transformation mechanism of beta-tricalcium phosphate (beta-TCP). X-ray diffraction analysis shows that the development of beta-TCP is not via direct reaction between Ca and P for all the different combinations between Ca and P precursors. The activation energy of beta-TCP formation with (NH4)H2PO4 as precursor was higher than that with H3PO4. Following the Johnson-Mehl-Avrami equation, the reaction kinetics of beta-TCP phase formation is found one-dimension growth with interface-controlled and diffusion controlled growth depending on the annealing temperature. There exists a transition between 750 degrees C and 825 degrees C, and the transition rate from interface-controlled to diffusion-controlled growth is precursor-dependent.

Apatites↗