Synthesis and dispersion of hydroxyapatite nanopowders

Spherical, rod and fibroid hydroxyapatite [HAp, Ca₁₀(PO₄)₆(OH)₂] nanoparticles were prepared and dispersed in aqueous media. Temperature and solution pH were the key factors to synthesis of different morphology and crystallinity. Processing conditions were selected from ternary diagram of pH, temperature and Ca:P ratio. High hydroxyl ion concentration (12.25 ≥ pH ≥ 10.5) and low temperature (298 K) favored isotropic non-confined spherical particles, intermediate concentration (9.5 ≥ pH ≥ 7.75) and low temperature (303 K) initiated the anisotropic growth of rod shaped particles but low concentration (7 ≥ pH ≥ 5.25) and high temperature (353 K) accelerated one-dimensional fibroid morphology. The dispersed HAp–citrate complex exhibited a constant zeta potential and size distribution for six months.     

Effects of pH and temperature on microstructure and morphology of hydroxyapatite/collagen composites synthesized in vitro

Hydroxyapatite/collagen composites (HAp/Col) were synthesized, in vitro, using the self-organization mechanism, by the co-precipitation of collagen, extracted from pork skin, and aqueous H₃PO₄ and Ca(OH)₂. The effects of pH and temperature, on the microstructure and morphology of HAp/Col composites, were extensively studied.It was shown that the yield of the composite is closely related to the pH value, during preparation. At high pH, the white precipitate of the composites was formed in a large quantity. In contrast, the yield was quite low, at low pH, owing to the dissolution of the composite precipitates. The electrophoretogram of the as-prepared composite shows that the α1(I) chain was dominant and that there was no protein residue in the supernatant of the reaction solution. Accordingly, it is reasonable to assume that the collagen molecules had completely reacted with hydroxyapatite.A closer look at the structural evolution of HAp/Col composites revealed that, at pH = 5, there was no sign of interfacial interaction between hydroxyapatite and collagen, during the first 36 h. In contrast, under alkaline conditions, the amount of α(I) chains was indeterminate, at the start of precipitation, while the concentration of β-chains decreased gradually. The chemical structure of HAp/Col, as determined by FTIR spectra, revealed that the bending vibration of phosphate contours can be attributed to the dissolution of HAp, at low pH. The crystalline of HAp was readily discerned, for all samples, as revealed by the XRD patterns. The morphology of the HAp/Col, prepared under neutral to alkaline conditions (7 ≤ pH ≤ 9), exhibited a compact, coral reef-like structure. In summary, HAp/Col composites can be synthesized, in vitro, under conditions native to animal physiology. The desirable conditions for synthesis are a higher pH (8 to 9) and a temperature of 40 °C.     

Nanocrystalline hydroxyapatite bioceramic using microwave radiation: Synthesis and characterization

In this work, we synthesized bioactive hydroxyapatite (Ca₁₀(PO₄)₆(OH)₂, HAp) ceramic powder in the lower-end of nano-regime using microwave radiation, which offers several advantages. The powder was synthesized using calcium nitrate tetrahydrate and sodium phosphate dibasic anhydrous as the starting materials. EDTA served as the complex reagent. The pH of the final suspension was adjusted to 9 by adding ammonium hydroxide. Applied microwave power of 600 W, pH of the suspension, mole ratio of Ca/P in the staring chemicals, and the chelating effect of EDTA served as the factors in the synthesis of nanocrystalline HAp powder. The synthesized powder was studied using various characterizing techniques viz., XRD, SEM, HR-TEM, EDS, TG/DTA and FT-IR to determine powder morphology, particle-size, crystallinity, phases, elemental composition and thermal behavior. Results confirmed highly crystalline nano-powder (5–30 nm) with elemental composition of Ca and P in HAp phase and possessed mixed (elliptical and rod-shape) morphology. Using the Scherrer formula, the average crystallite size was found to be 12 nm. The FT-IR confirmed that the powder is of typical apatite structure. Thermal analysis showed a remarkably lower initial dehydroxylation temperature, compared to micron sized HAp, as reported in literature.     

Ribbon-like and rod-like hydroxyapatite crystals deposited on titanium surface with electrochemical method

Homogeneous coatings were attained by electrochemical method in electrolytes containing Ca²⁺ and PO₄³⁻ ions with Ca/P ratio being 1.67. SEM observation showed that the hydroxyapatite (HAp,Ca₁₀(PO₄)₆(OH)₂) crystals prepared with higher concentration electrolyte (4 × 10^− 2 M Ca²⁺) are ribbon-like with thickness of nanometer size, a morphology seldom reported previously. In an electrolyte of lower concentration (6 × 10^− 4 M Ca²⁺), the HAp crystals formed are rod-like with a hexagonal cross section and diameter of about 70–80 nm. XRD patterns and IR spectra confirmed that the coatings consist of HAp crystals. TEM micrographs and SAD indicated that the longitude direction for both ribbon-like and rod-like crystal is [002], and the flat surface of the ribbon is (110). HRTEM showed that the ribbon-like crystal is a mixture of HAp and octacalcium phosphate (OCP, Ca₈H₂(PO₄)₆.5H₂O).     

Bone cell–material interactions on metal-ion doped polarized hydroxyapatite

The objective of this work is to study the influence of Mg²⁺ and Sr²⁺ dopants on in vitro bone cell–material interactions of electrically polarized hydroxyapatite [HAp, Ca₁₀(PO₄)₆(OH)₂] ceramics with an aim to achieve additional advantage of matching bone chemistry along with the original benefits of electrical polarization treatment relevant to biomedical applications. To achieve our research objective, commercial phase pure HAp has been doped with MgO, and SrO in single, and binary compositions. All samples have been sintered at 1200 °C for 2 h and subsequently polarized using an external d.c. field (2.0 kV/cm) at 400 °C for 1 h. Combined addition of 1 wt.% MgO/1 wt.% SrO in HAp has been most beneficial in enhancing the polarizability in which stored charge was 4.19 μC/cm² compared to pure HAp of 2.23 μC/cm². Bone cell–material interaction has been studied by culturing with human fetal osteoblast cells (hFOB) for a maximum of 7 days. Scanning electron microscope (SEM) images of cell morphology reveal that favorable surface properties and dopant chemistry lead to good cellular adherence and spreading on negatively charged surfaces of both Sr²⁺ and Mg²⁺ doped HAp samples over undoped HAp. MTT assay results at 7 days show the highest viable cell densities on the negatively charged surfaces of binary doped HAp samples, while positive charged doped HAp surfaces exhibit limited cellular growth in comparison to neutral surfaces.     

The impact of Cu(II) ions doping in nanostructured hydroxyapatite powder: A finite element modelling study for physico-mechanical and biological property evaluation

The synthesis of doped nanostructured materials with multifunctional properties and improved biocompatibility have immense potential for biomedical applications. In this present study, a facile wet chemical precipitation method was employed to synthesize hydroxyapatite (HAp) and different concentrations copper doped HAp, and Cu_x-HAp (x = 1, 2, and 4 mol%) nano materials. Sophisticated analytical and spectroscopic techniques were employed to confirm its physico-chemical properties, and morphological features. The synthesized HAp, Cu_x-HAp were further studied as a drug nanocarrier using doxorubicin hydrochloride (DOX) as a model drug, which results a maximum drug release of ～34.3% (at pH 4.5) for 1 mol% of Cu-HAp. The nanostructured materials were further used to fabricate scaffolds by employing gel-casting method. The finite element modeling theoretical approach was adopted, to correlate the force distribution over the developed scaffold during mechanical characterization. The in vitro study confirmed the nontoxic behavior of the HAp and Cu_x-HAp scaffolds using MG-63 cell line. The developed scaffold effectively facilitates and simulates the new cell attachment, growth, and proliferation on its surface with adequate (～7.87 MPa) compressive strength properties. The enhanced biocompatibility with improved mechanical stability of Cu_x-HAp nanomaterials could address some of the critical challenges in biomedical applications.     

In vitro biocompatibility and antimicrobial activity of wet chemically prepared Ca10−xAgx(PO4)6(OH)2 (0.0 ≤ x ≤ 0.5) hydroxyapatites

Herein, we report the effect of silver ions on the physical, antimicrobial and cytocompatibility properties of wet chemically synthesized silver doped Ca_10?xAg_x(PO₄)₆(OH)₂ (0.0 ≤ x ≤ 0.5) hydroxyapatites (HAp). Silver ions containing HAp exhibit the comparable density, hardness and enhanced antimicrobial properties, in comparison to parent HAp. The optical absorption measurements confirm the presence of silver ions in the doped compositions, which are responsible for as increased antimicrobial property of doped HAp materials for x > 0.3. The cytotoxicity behavior of the doped HAp was evaluated using mouse fibroblast (L929) cell line. The important result has been that doped HAp (x > 0.3) exhibit statistically (significant) lower cell viability in comparison to undoped HAp. However, no difference in cellular functionality on doped HAp surfaces, in terms of cell adhesion and proliferation could be qualitatively observed in reference to undoped HAp. In order to explain the observed antimicrobial and cell viability properties, the in vitro release of Ag⁺ ions has been quantified using Inductively Coupled Plasma-Atomic Emission Spectroscopy (ICP-AES) and solubility was measured by weight loss in acetate buffer solution.     

Structural and microstructural characterizations of nanocrystalline hydroxyapatite synthesized by mechanical alloying

Single phase nanocrystalline hydroxyapatite (HAp) powder has been synthesized by mechanical alloying the stoichiometric mixture of CaCO₃ and CaHPO₄ powders in open air at room temperature, for the first time, within 2 h of milling. Nanocrystalline hexagonal single crystals are obtained by sintering of 2 h milled sample at 500 °C. Structural and microstructural properties of as-milled and sintered powders are revealed from both the X-ray line profile analysis and transmission electron microscopy. Shape and lattice strain of nanocrystalline HAp particles are found to be anisotropic in nature. Particle size of HAp powder remains almost invariant up to 10 h of milling and there is no significant growth of nanocrystalline HAp particles after sintering at 500 °C for 3 h. Changes in lattice volume and some primary bond lengths of as-milled and sintered are critically measured, which indicate that lattice imperfections introduced into the HAp lattice during ball milling have been reduced partially after sintering the powder at elevated temperatures. We could achieve ~ 96.7% of theoretical density of HAp within 3 h by sintering the pellet of nanocrystalline powder at a lower temperature of 1000 °C. Vickers microhardness (VHN) of the uni-axially pressed (6.86 MPa) pellet of nanocrystalline HAp is 4.5 GPa at 100 gm load which is close to the VHN of bulk HAp sintered at higher temperature. The strain-hardening index (n) of the sintered pellet is found to be > 2, indicating a further increase in microhardness value at higher load.     

Effect of urea on formation of hydroxyapatite through double-step hydrothermal processing

The effect of urea on the formation of hydroxyapatite (HAp) was studied by employing the double-step hydrothermal processing of a powder mixture of beta-tricalcium phosphate (β-TCP) and dicalcium phosphate dihydrate (DCPD). Co-existence of urea was found to sustain morphology of HAp crystals in the compacts under an initial concentration of 2 mol dm^-3 and less. Homogenous morphology of needle-like crystals was observed on the compacts carbonated owing to decomposition of urea. Carbonate ions (CO₃^2-) was found to be substituted in both the phosphate and hydroxide sites of HAp lattice. The synthesized HAp was calcium deficient, as it had a Ca/P atomic ratio of 1.62 and the phase was identified as calcium deficient hydroxyapatite (CDHA). The release of CO₃^2- ions from urea during the hydrothermal treatment determined the morphology of the CDHA in the compacts. The usage of urea in the morphological control of carbonate-substituted HAp (CHAp) employing the double-step hydrothermal method is established.     

Preparation of high strength macroporous hydroxyapatite scaffold

Hydroxyapatite (HAp) powder was prepared from CaNO₃·4H₂O and (NH₄)₂HPO₄ by wet-chemical method and has phase stable up to 1250 °C. High strength macroporous HAp–naphthalene (HN) and HAp–naphthalene–benzene (HNB) scaffolds were fabricated by adapting sintering method. The resulting HAp scaffolds have porosity about 60 vol.% with compressive strength of ~ 11 MPa and average pore diameter in the range of ~ 125 μm. The incorporation of benzene in HN scaffold reduces the strength whereas enhanced both the porosity and pore size distribution. XRD, FTIR, SEM and mercury porosimeter techniques were used to study the phase purity, morphology, pore size and pore size distribution of scaffold. The study compared the effect of concentration of naphthalene on strength, porosity and pore size distribution on both HN and HNB scaffold. In-vitro bioactivity studies on HN and HNB scaffolds show the nucleation of spherical carbonated apatite particles on the surface in SBF solution.     

Fabrication of HAp–8YSZ composite layer on Ti/TiO2 nanoporous substrate by EPD/MAO method

Zirconia/Hydroxyapatite composites containing 20–50 wt.% 8YSZ were prepared on Ti/TiO₂ substrates by electrophoretic deposition (EPD)/micro-arc oxidation (MAO) process. Titania, as an inner layer, was grown on the Ti plates using MAO treatment in order to form a strong join between substrate and HAp. These composites were produced by EPD in ethanol containing ZrO₂/HAp particles at 50, 100 and 150 V in 1 min. As-prepared samples were sintered at 900, 1100 and 1300 °C. HAp, β-TCP, CaZrO₃ phases were identified using X-ray diffractometry analysis (XRD). Scanning electron microscopy (SEM) utilized to study the surface morphology indicated a crack free microstructure at 1300 °C.     

Microwave-assisted hydrothermal synthesis of zinc oxide particles starting from chloride precursor

Zinc oxide (ZnO) was synthesized using a microwave assisted hydrothermal (MAH) process based on chloride/urea/water solution and under 800 W irradiation for 5 min. In the bath, Zn²⁺ ions reacted with the complex carbonate and hydroxide ions to form zinc carbonate hydroxide hydrate (Zn₄CO₃(OH)₆·H₂O), and the conversion from Zn₄CO₃(OH)₆·H₂O to ZnO was synchronously achieved by a MAH process. The as-prepared ZnO has a sponge-like morphology. However, the initial sponge-like morphology of ZnO could change to a net-like structure after thermal treatment, and compact nano-scale ZnO particles were finally obtained when the period of thermal treatment increased to 30 min. Pure ZnO nanoparticles was obtained from calcination of loose sponge-like ZnO particles at 500 °C. The analysis of optical properties of these ZnO nanoparticles showed that the intensity of 393 nm emission increased with the calcination temperature because the defects were reduced and the crystallinity was improved.     

Biomimetic mineralization of calcium phosphate crystals in polyacrylamide hydrogel: Effect of concentrations of calcium and phosphate ions on crystalline phases and morphology

Calcium phosphate crystals were synthesized in polyacrylamide (PAAm) hydrogel, and the effects of the concentrations of calcium and phosphate ions on the crystalline phases and morphology were investigated. PAAm hydrogels containing diammonium hydrogen phosphate ((NH₄)₂HPO₄) were transformed into calcified materials by diffusion of calcium ions from calcium nitrate (Ca(NO₃)₂) aqueous solution into the gels. Several kinds of calcium phosphate crystals were precipitated at various Ca(NO₃)₂ concentrations (0.5–4.0 mol·dm^? 3), or (NH₄)₂HPO₄ contents (3.6–21.6 mmol) in the gels. The crystalline phases were mainly determined by the (NH₄)₂HPO₄ content in the gels. When the (NH₄)₂HPO₄ content was ≥ 10.8 mmol, hydroxyapatite (HAp) formed near the interfaces between Ca(NO₃)₂ solution and the gels, whereas octacalcium phosphate (OCP) formed in gels with ≤ 10.8 mmol (NH₄)₂HPO₄. HAp crystals were granular in form and about 200 nm in diameter, and OCP crystals were spherulitic with diameter 10–70 μm.     

Hierarchical architecture of Bi12TiO20 via ethylene glycol-mediated synthesis route

Hierarchical architecture of cubic sillenite bismuth titanate (Bi₁₂TiO₂₀) is successfully synthesized using simple ethylene glycol-mediated self-assembly; followed by calcination under air at 600 °C for 30 min. The products are characterized using field emission scanning electron microscopy (FE-SEM), fourier transform infrared spectroscopy (FT-IR), thermogravimetric-differential thermal analysis (TG-DTA), and X-ray diffraction (XRD). The reaction time has an effect on the product morphology, which changed from a leaf- and circular plate-like structure to a completely formed hierarchical structure within 2 h. The hierarchical structure of Bi₁₂TiO₂₀, having a size around 3 μm, is composed of 2D twist nanoplates, and each twist nanoplate has a thickness around 10–30 nm.     

Influence of temperature, [Ca2+], Ca/P ratio and ultrasonic power on the crystallinity and morphology of hydroxyapatite nanoparticles prepared with a novel ultrasonic precipitation method

Nanocrystalline hydroxyapatite was prepared by a precipitation method with the aid of ultrasonic irradiation using Ca(NO₃)₂ and NH₄H₂PO₄ as source material and carbamide (NH₂CONH₂) as precipitator. The influence of Ca/P molar ratio, precipitation temperature, concentration of Ca²⁺ ([Ca²⁺]) and ultrasonic power on the crystallinity of the nanopowder were systematically investigated by XRD analysis. The size of the as-prepared particles was analyzed using TEM and XRD methods. The results revealed that the monophase hydroxyapatite could be obtained at the following technological conditions: [Ca²⁺] = 0.01–0.1 mol/L, ultrasonic power = 300 W, Ca/P (mol) = 1.2–2.5 and T = 313–353 K. In addition, the acicular and spherical particles could be prepared at different ultrasonic powers of 300 and 200 W, respectively.     

Strontium doped hydroxyapatite film formed by micro-arc oxidation

A porous strontium-doped hydroxyapatite (Sr-HA) film was prepared on titanium substrates by an electrochemical oxidation method, i.e. micro-arc oxidation (MAO). The reaction was processed using a pulse power supply with titanium substrate acted as the anode in electrolytic solution containing calcium acetate, strontium acetate and β-glycerol phosphate disodium salt pentahydrate (β-GP). The thickness, phase, composition and morphology of the coatings were investigated with X-ray diffraction (XRD), electron probe microanalysis (EPMA) and scanning electron microscopy (SEM) with energy dispersive X-ray spectrometer (EDS). The thickness of the film was about 20 µm with the porous and uneven surfaces. XRD showed that the film was mainly made up of hydroxyapatite doped with strontium and the (Sr/(Sr + Ca) ratios ranged from 0–100%, which could be expressed as the general formula of Ca_10 ? XSr_X(PO₄)₆(OH)₂, 0 ≤ X ≤ 10). Such films are expected to have significant medical applications as dental implants and artificial bone joints.     

Adsorption of gum Arabic on bioceramic nanoparticles

Surface modification agents can be used to tailor the surface chemistry and biological activity of bioceramic nanoparticles in very intriguing ways. However, the specific modes of interactions between macromolecules and nanoparticles can be difficult to characterize. The aim of this study was to investigate the adsorption of gum Arabic on hydroxyapatite (HAp) and magnetic nanoparticles (MNP) using the bicinchoninic acid (BCA) test. Gum Arabic (GA) is a natural gum that has been widely used as an emulsifying agent and shows promise for dispersing nanoparticles in aqueous solutions. The adsorption of GA onto HAp nanoparticles followed a Langmuir isotherm with an adsorption plateau occurring at 0.2 g GA/g HAp. The adsorption of GA onto MNP attained a maximum value of 0.6 g GA/g MNP, after which it decreased to approximately 0.2 g GA/g MNP. The maximum adsorption density of GA on both MNP and HAp is equivalent when normalized to the specific surface area (4 × 10^− 3 g GA/m²). Adsorbed GA molecules were displaced from the surface of HAp and MNP in the presence of phosphate ions.     

In-vitro bioactivity,biocorrosion and antibacterial activity of silicon integrated hydroxyapatite/chitosan composite coating on 316 L stainless steel implants

A simple and effective ultrasonication method was applied for the preparation of 0, 0.4, 0.8, 1.0 and 1.6 wt% silicon substituted hydroxyapatite (HAp) (SH). The Ca/P ratio of the synthesised SH nanoparticles were in the range of 1.58–1.70. Morphological changes were noticed in HAp with respect to the amount of Si from 0 to 1.6 wt%. The morphology of the particles changed from spherical shape to rod-like morphology with respect to the amount of Si which was confirmed using transmission electron microscopy. X-ray diffraction studies confirm the formation of phase pure SH nanoparticles without any secondary phase. Chitosan (CTS) blended SH nanocomposites coating on surgical grade 316 L stainless steel (316 L SS) implant was made by spin coating technique. The surface of the coated implant was characterised using scanning electron microscopy which confirms the uniform coating without cracks and pores. The increased corrosion resistance of the 1.6 wt% of SH/CTS-coated SS implant in the simulated body fluid (SBF) indicates the long-term biostability of SH composite-coated ceramics in vitro than the 0 wt% SH/CTS. The testing of SH/CTS nanocomposites with gram-positive and gram-negative bacterial strains confirms that the antibacterial ability improves with the higher substitution of Si. In addition, formation of bone-like apatite layer on the SH/CTS-coated implant in SBF was studied through SEM analysis and it confirms the ability to increase the HAp formation on the surface of 1.0 wt% SH/CTS-coated 316 L SS implant.     

Antibacterial effects of silver-doped hydroxyapatite thin films sputter deposited on titanium

Since many orthopedic implants fail as a result of loosening, wear, and inflammation caused by repeated loading on the joints, coatings such as hydroxyapatite (HAp) on titanium with a unique topography have been shown to improve the interface between the implant and the natural tissue. Another serious problem with long-term or ideally permanent implants is infection. It is important to prevent initial bacterial colonization as existing colonies have the potential to become encased in an extracellular matrix polymer (biofilm) that is resistant to antibacterial agents. In this study, plasma-based ion implantation was used to examine the effects of pre-etching on plain titanium. Topographical changes to the titanium samples were examined and compared via scanning electron microscopy. Hydroxyapatite and silver-doped hydroxyapatite thin films were then sputter deposited on titanium substrates etched at ? 700 eV. For silver-doped films, two concentrations of silver (~ 0.5 wt.% and ~ 1.5 wt.%) were used. Silver concentrations in the film were determined using energy dispersive X-ray spectroscopy. Hydroxyapatite film thicknesses were determined by measuring the surface profile using contact profilometry. Staphylococcus epidermidis and Pseudomonas aeruginosa adhesion studies were performed on plain titanium, titanium coated with hydroxyapatite, titanium coated with ~ 0.5 wt.% silver-doped hydroxyapatite, and titanium coated with ~ 1.5 wt.% silver-doped hydroxyapatite. Results indicate that less bacteria adhered to surfaces containing hydroxyapatite and silver; further, as the hydroxyapatite films delaminated, silver ions were released which killed bacteria in suspension.     

Sodium niobate particles with controlled morphology synthesized by hydrothermal method and their use as templates in KNN fibers

Sodium niobate – NN (NaNbO₃) powders were synthesized by hydrothermal process to be used as template particles in the fabrication of textured lead free piezoelectric ceramics. Sodium hexaniobate–Na₈Nb₆O₁₉⋅13H₂O particles with rod-like morphology were synthesized at 120 °C. Particles with needle-like morphology and Na₂(Nb₂O₆)(H₂O) phase started to form at temperatures of 130 °C and above. Synthesis at 150 °C yields particles with totally needle-like morphology and consisting entirely of the Na₂(Nb₂O₆)(H₂O) phase. Sodium niobate–NaNbO₃ particles with cubic morphology were synthesized at temperatures of 160 °C and above. Rod-like and needle-like morphology was retained even after annealing at 400 °C for 1 h. A preliminary study was also done to integrate these anisometric template particles in the preparation of textured potassium sodium niobate (KNN) fibers.