Calcium Deficient Carbonated Apatite Research Articles

Fabrication of Hydroxyapatite/Tantalum Composites by Pressureless Sintering in Different Atmosphere.

The effect of sintering atmosphere (air and Ar) and temperature (1100, 1200, 1300 °C) on the microstructure, mechanical properties, and bioactivity of hydroxyapatite/tantalum (HA/Ta) composites were systematically investigated by pressureless sintering of the mixture of HA and Ta powders. It shows that the sintering atmosphere greatly impacts the phase composition and microstructure of the HA/Ta composites. The higher diffusion of atoms promotes shrinkage and causes deeper reaction fusion between the HA matrix and Ta, which improved the interfacial binding of the HA/Ta composites. The refined grain structure and improved interfacial binding obtained within the Ar atmosphere compared to the air atmosphere benefit the mechanical properties. The maximum bending strength and shrinkage observed for the composites sintered at 1300 °C in the Ar atmosphere are 27.24 MPa and 6.65%, respectively. The cell counting kit-8 (CCK-8) method was used to investigate the in vitro cytocompatibility of HA/Ta composites. The results revealed that the HA/Ta composites sintered with different conditions have no cytotoxicity. The simulated body fluid (SBF) soaking results showed that all of the studied composites possess desirable bioactivity, as demonstrated by their ability to form calcium-deficient carbonate apatite layer on the surfaces. For composites sintered at 1300 °C, the surface apatite layer coverage of the composites obtained in the Ar atmosphere was increased by 139.7% than the ones obtained in air, which confirmed an enhanced bioactive mineralization ability. The results indicated that the HA/Ta composites sintered at 1300 °C in Ar possess desirable mechanical properties and bioactivity. This work opens up the new possibility for preparing HA-based composites and is of great value in biomedical applications.

Low-temperature synthesis of nanometric apatite from biogenic sources

Nanosized calcium deficient carbonated apatite was synthesized from three different natural resources, namely chicken eggshells (calcite), cuttlefish bones (aragonite) and mussel shells (aragonite/calcite). The calcium precursors were ball-milled in (NH4)2HPO4 or H3PO4 – containing aqueous medium for different times and then dried at temperatures ranging from 20 °C to 150 °C. The formation of hydroxyapatite (HA) is shown to be strongly affected by such treatment, the amount of synthesized HA increasing with temperature. Aragonite from cuttlebones and the aragonitic fraction of mussels is found to be transformed more easily with respect to the calcite from eggshells. The reaction is also favored by an acidic milling media with respect to basic one. The feasibility of the synthesis process at nearly room temperature described in the present work has interesting potentialities in the retention of the organic portions of the original biological resource in the produced nanometric inorganic compound.

Biomimetic deposition of carbonate apatite and role of carbonate substitution on mechanical properties at nanoscale

This work reports on the role of carbonate substitution on the mechanical properties of calcium deficient carbonate apatite which resembles closely the structure of bone apatite. The carbonate content determined using vibrational spectroscopy suggests two stage substitutions. The residual stress is evaluated using non-destructive X-ray diffraction procedure for these polycrystalline aggregates. The crystallinity increases with deposition time and increased carbonate substitution does not impede the crystal growth and no phase change is observed. A carbonate content of 6.34wt. % corresponds to a minimum residual stress without compromising the crystallinity of the films. The carbonate substitution might be associated with the calcium vacancy which is considered as the thermodynamically feasible mechanism for B-type carbonate apatite.

In vitro assessment of zinc apatite coatings on titanium surfaces

In this paper, coatings of hydroxyapatite partially substituted with zinc (ZnHA) were produced on titanium substrates by a two-step hydrothermal process using a precursor solution rich in calcium, phosphate and zinc. Activation of titanium surfaces was performed by oxidation with an acidic HF/HNO3 solution. The coated substrates were then converted into HA by immersion in an alkali 0.1M NaOH solution. The ZnHA samples were characterized by several techniques and their in vitro behavior was studied in comparison to hydroxyapatite (HA) and titanium (Ti-control) samples. A uniform and homogeneous calcium-deficient carbonate apatite coating was obtained for all samples, both doped and undoped with zinc. The percentage of zinc incorporated in the coatings is 7at%, and the Ca/P ratio is 1.61(±0.01) for both types of samples, suggesting that Zn is incorporated substitutionally, replacing Ca atoms into the HA structure. The incorporation of Zn in the HA structure changed the crystals morphology, reduced crystals sizes and decreased the deposition rate showing that zinc is an inhibitor of the growth of HA crystal. X-ray diffraction showed that HA is the single crystalline phase present after alkali treatment. The coating adhesion strength was evaluated in terms of the critical load (Lc) obtained from scratch tests and no significant difference was found between the two tested groups, indicating the good adhesion of ZnHA to Ti substrates. The in vitro response of human osteoblasts (HOB) exposed to the surfaces of HA and ZnHA coatings was evaluated. The results of Live/Dead tests showed cell viability for all samples surfaces, but the adhesion and proliferation tests showed that ZnHA samples presented better adhered and spread cells compared with HA. ZnHA coatings presented cells with elongated or polygonal shapes and clearly more spread than HA. Quantitative analysis showed that there was a significantly higher number of cells adhered to ZnHA coatings compared to HA, indicating the zinc incorporation stimulates osteoblast proliferation.

Polyelectrolyte multilayer-calcium phosphate composite coatings for metal implants.

The preparation of organic-inorganic composite coatings with the purpose to increase the bioactivity of bioinert metal implants was investigated. As substrates, glass plates and rough titanium surfaces (Ti-SLA) were employed. The method comprises the deposition of polyelectrolyte multilayers (PEMLs) followed by immersion of the coated substrate into a calcifying solution of low supersaturation (MCS). Single or mixed PEMLs were constructed from poly-L-lysine (PLL) alternating with poly-L-glutamate, (PGA), poly-L-aspartate (PAA), and/or chondroitin sulfate (CS). ATR-FTIR spectra reveal that (PLL/PGA)10 multilayers and mixed multilayers with a (PLL/PGA)5 base contain intermolecular β-sheet structures, which are absent in pure (PLL/PAA)10 and (PLL/CS)10 assemblies. All PEML coatings had a grainy topography with aggregate sizes and size distributions increasing in the order: (PLL/PGA)n < (PLL/PAA)n < (PLL/CS)n. In mixed multilayers with a (PLL/PGA)n base and a (PLL/PAA)n or (PLL/CS)n top, the aggregate sizes were greatly reduced. The PEMLs promoted calcium phosphate nucleation and early crystal growth, the intensity of the effect depending on the composition of the terminal layer(s) of the polymer. In contrast, crystal morphology and structure depended on the supersaturation, pH, and ionic strength of the MCS, rather than on the composition of the organic matrix. Crystals grown on both uncoated and coated substrates were mostly platelets of calcium deficient carbonate apatite, with the Ca/P ratio depending on the precipitation conditions.

Hydroxyapatite/sericin composite film prepared through mineralization of flexible ethanol-treated sericin film with simulated body fluids

Abstract A flexible ethanol-treated sericin film was prepared and used to form a hydroxyapatite/sericin composite film by mineralizing it with simulated body fluids solution. The hydroxyapatite mineral deposited on the surface of the ethanol-treated sericin film was characterized by Scanning Electron Microscopy, Fourier Transform Infrared Spectroscopy, Energy Dispersive X-Ray Spectroscopy, and X-ray diffraction analysis. Results indicated that the quantity of deposited minerals increased with increasing mineralization time and showed a three-dimensional structure. This structure was determined to be calcium-deficient carbonate apatite with poor crystallinity and growth in the direction of the c -axis similar to natural bone mineral. The mechanical properties of the hydroxyapatite/sericin composite film decreased with increasing mineralization time. XTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) tests showed that the hydroxyapatite/sericin composite film promoted cell viability, which is beneficial for its application in bone tissue engineering.

Calcium Phosphate Cement (CPC): A Critical Development Path

The first generation of synthetic bone substitute materials (BSM) was initially investigated in the mid 1970s using hydroxyapatite (HA) as a biomaterial for remodeling of bone defects. The concepts established by CPC pioneers in the early 1980s were used as a platform to initiate a second generation of BSM for commercialization. Since then, advances have been made in composition, performance and manufacturing. A self-setting and injectable calcium phosphate cement (CPC) based on amorphous calcium phosphate (ACP) with calcium to phosphate (Ca/P) atomic ratio less than 1.5, combined with dicalcium phosphate dihydrate (DCPD or brushite, seeded with apatite), is proposed. Amorphization of raw material was observed following high energy mechano-chemical processing. Upon hydration, the cement hardened in less than 3 minutes at 37°C and reached a maximum compressive strength of about 50 MPa. The final product was a low crystalline calcium deficient carbonated apatite similar to the composition and structure of bone mineral. In vivo performance of this cement in mediating bone healing was compared to α-BSM® in a rabbit femoral defect model. Performance characteristics of some commercially available CPC products are compared. The concerns of CPC designers and the needs of product users (surgeons) are discussed.

Surface functional group dependent apatite formation on bacterial cellulose microfibrils network in a simulated body fluid

The apatite forming ability of biopolymer bacterial cellulose (BC) has been investigated by soaking different BC specimens in a simulated body fluid (1.5 SBF) under physiological conditions, at 37 degrees C and pH 7.4, mimicking the natural process of apatite formation. From ATR-FTIR spectra and ICP-AES analysis, the crystalline phase nucleated on the BC microfibrils surface was calcium deficient carbonated apatite through initial formation of octacalcium phosphate (OCP) or OCP like calcium phosphate phase regardless of the substrates. Morphology of the deposits from SEM, FE-SEM, and TEM observations revealed the fine structure of thin film plates uniting together to form apatite globules of various size (from <1 mum to 3 mum) with respect to the substrates. Surface modification by TEMPO (2,2,6,6-tetramethylpyperidine-1-oxyl)-mediated oxidation, which can readily form active carboxyl functional groups upon selective oxidation of primary hydroxyl groups on the surface of BC microfibrils, enhanced the rate of apatite nucleation. Ion exchanged treatment with calcium chloride solution after TEMPO-mediated oxidation was found to be remarkably different from other BC substrates with the highest deposit weight and the smallest apatite globules size. The role of BC substrates to induce mineralization rate differs according to the nature of the BC substrates, which strongly influences the growth behavior of the apatite crystals.

Properties of Calcium Phosphate Powder Prepared from Phosphoryl Oligosaccharides of Calcium

The phosphoryl oligosaccharides of calcium (POC), extracted from potato starch, are composed of phosphorus oligosaccharides and calcium ions. Ultrafine calcium phosphate particles, whose main phase was hydroxyapatite (Ca10(PO4)6(OH)2: HAp), could be prepared through the hydrothermal treatment of POC solution at a temperature between 110 and 130°C; X-ray diffraction indicated the crystallinity of HAp in the resulting powder to be poor and similar to that of living bone. The present HAp powder was regarded to be calcium deficient carbonate apatite with the OH- group being partly substituted by a carbonate (CO3 2-) group. The solubility of the resulting powder in dilute hydrochloric acid was higher compared to that of commercially available HAp, suggesting excellent bioabsorbability for the present powder.

Inherent Luminescence of Annealed Biomimetic Apatites

Biomimetic apatite coatings are widely used in orthopaedic applications to provide bioinert material surfaces with bioactive behaviour by means of initiating bone growth at the implant surface. In this study we manufactured biomimetic calcium phosphate coatings consisting of a calcium deficient carbonated apatite by immersing activated titanium platelets into simulated body fluid (SBF). The development of the crystal phases was monitored by X-ray diffractometry (XRD) in addition to Fourier-transform infrared (FT-IR) spectroscopy. After annealing in air up to 600 °C luminescence of the biomimetically derived apatite was observed. The photo-induced emission spectra were recorded in the range from 400-750 nm at excitation wavelengths ranging 238 to 450 nm. A blue (437 nm) and a green (556 nm) emission were found between 200 to 600 °C visually appearing white. The results are discussed in terms of chemical and crystallographic changes in the biomimetic calcium phosphate layer during heat treatment.

Photoluminescence of annealed biomimetic apatites

Biomimetic apatite coatings are widely used in orthopaedic applications to provide bioinert material surfaces with bioactive behaviour by means of initiating bone growth at the implant surface. In this study we manufactured biomimetic calcium phosphate coatings consisting of a calcium deficient carbonated apatite by immersing activated titanium platelets into simulated body fluid. The development of the crystal phases was monitored by X-ray diffractometry in addition to Fourier-transform infrared spectroscopy. The microstructure of the biomimetic apatites and phase composition was analysed using scanning and transmission electron microscopy as well as attached energy dispersive X-ray spectrometry. The samples were annealed in air yielding in an inherent luminescence of the biomimetic apatite up to temperatures of 600 °C. The photo-induced emission spectra were recorded in the range from 400 to 750 nm at excitation wavelengths ranging 310–450 nm. A blue (437 nm) and a green (561 nm) emission were found between 200 and 600 °C visually appearing white. Photoluminescence of annealed biomimetic apatites might be of interest for histological probing and monitoring of bone re-modelling. The results are discussed in terms of chemical and crystallographic changes in the calcium phosphate layer during heat treatment.

Comparison between the in vitro surface transformations of AP40 and RKKP bioactive glasses

Two bioactive silica-phosphate glasses, AP40 and RKKP, were compared in their behaviour in simulated biological environment. Their chemical composition is practically identical, except that RKKP contains small amounts of amphoteric network-former oxides Ta2O5 and La2O3 (composition in wt% for AP40: beta-Ca3(PO4)2 24.50, SiO2 44.30, CaO 18.60, Na2O 4.60, K2O 0.19, MgO 2.82, CaF2 4.99; RKKP: beta-Ca3(PO4)2 24.23, SiO2 43.82, CaO 18.40, Na2O 4.55, K2O 0.19, MgO 2.79, CaF2 4.94, Ta2O5 0.99, La2O3 0.09). Previous investigations showed a better performance in osteopenic bone for RKKP. To gain more insight into these differences in biological behaviour, the in vitro bioactivity of the glasses was studied by treatment with a continuously replenished Hanks' Balanced Salt Solution (HBSS). The glasses were examined before and after HBSS treatment for 20 and 40 days by X-ray Diffraction (XRD), Scanning Electron Microscopy (SEM), X-ray Energy Dispersion (EDX), Raman and IR vibrational spectroscopies. Some slight but notable differences between the two glasses were observed after HBSS treatment. IR and EDX analyses showed that deposits formed on both glasses were composed of a calcium deficient carbonate-apatite; however, the layer formed on RKKP glass was found to be slightly more calcium deficient and thinner. EDX analysis evidenced the presence of a small percentage of F- ions only in the layers formed on the RKKP samples. The differences disclosed, although slight, can contribute to the understanding of the different biological behaviour previously observed.

Preparation of a bone-like apatite foam cement.

The preparation of a porous bone-like calcium deficient apatite implant material was investigated. A novel cement system composed of an equimolar mixture of Ca4(PO4)2O, Ca(HPO4)2 . H2O, and CaCO3 was used. At a liquid/powder ratio of 0.83 ml/g low density open framework foam cements were formed due to the rapid evolution of CO2. The initial product of the reactants was CaHPO4 . H2O which then reacted with Ca4(PO4)O, forming a calcium deficient carbonated apatite, upon soaking of the cement blocks in SBF. Foam-like cements were composed of a plate-like apatite due to epitaxial overgrowth and conversion of the brushite plate precursor. Cylinders of the foam cement were reinforced with an outer layer of a solid apatite cement to form a material suitable for application as a bone-section implant.