Crystalline HA Research Articles

Fabrication of strontium ions substituted hydroxyapatite from the shells of the golden apple snail (Pomacea canaliculate L) with enhanced osteoconductive and improved biological properties.

The use of biogenic calcium ions for the source of hydroxyapatite (HAp or HA) are very common and have been being explored extensively. However, it usually results high crystalline HA, due to high reaction and decomposition temperatures. In this study, strontium (Sr2+) doped HA from the golden apple snail shells (Pomacea canaliculate L) was successfully synthesized. It was indicated that Sr ions completely replaced calcium (Ca) ions, increased the lattice constant, and consecutively reduced HA crystallinity. Smaller crystal size and β-type carbonate (CO32-) ions substitution with Ca/P close to 1.67 molar ratio that mimic bone crystals were observed in Sr-doped HA, with significant increased rate of MC3T3-E1 cells viability and higher IC50 values. It was proven that Sr ions substitution resolved challenges on the use of biogenic sources for HA fabrication. Further in vivo study is needed to continue to valorise the results into real biomedical and clinical applications.

Structural and biophysical peculiarities of Nd2O3 doped borate bioglass

Different glasses with a variable composition of x Nd2O3.(46-x) B2O3.24.5 Na2O.26.5 CaO.2.6 P 2O5 (where X = 0–2.0 mol% of Nd2O3) were prepared and studied from the viewpoint of enhancing their bioactivities. All of the as-prepared glasses showed amorphous structures evidenced by the X-ray diffraction (XRD) patterns. On the other hand, XRD patterns of glasses soaked in sodium phosphate buffer solution revealed the formation of a variety of crystalline structures relevant to the formation of carbonated hydroxyapatite derivatives. According to the NMR data, the concentration of BO3 units increases at the expense of BO4 with increasing Nd2O3 content. Increasing of (BO3/BO4) molar ratio with increasing Nd2O3 plays the main role of enhancing the bioactivity of borate glasses. The percentage of BO3, BO4, and consequently N4 fraction (N4 =BO4/BO3+BO4) can simply be determined from the deconvolution process of FTIR and NMR spectra. A scanning electron microscope (SEM) and energy-dispersive x-ray (EDX) evaluation of the microstructures support the existence of crystalline HA after immersion. Both EDX and mapping of the sample after prolonged time intervals point to the homogeneity in the HA distribution of the studied sample.

Curcumin-loaded hydroxyapatite nanocomposite as a novel biocompatible shield for male Wistar rats from γ-irradiation hazard

Curcumin (CUR) is well known for its extraordinary benefits as an anti-cancer, anti-inflammatory, and wound healing agent. However, nano-formulation could maintain and regulate its pharmacological effect. Herein, we report the preparation of CUR/hydroxyapatite nanocomposite (CUR/HA NC) and its application in the protection of male Wistar rats from γ-irradiation carcinogenic consequences. TEM images of the nanocrystalline HA nanoparticles (NPs) had a rod-like form with average dimensions of 40±5 nm in length and 10 ± 5 nm in width. XRD analysis illustrated the formation of a single phase of hexagonal crystalline HA NPs. The presence of the CUR fingerprint is visible in its FTIR spectra of the CUR/HA NC. Biochemical analysis and histological examinations revealed that CUR/HA NC injection does not significantly affect non-irradiation rats compared to the control. However, when injected pre-irradiation, it controls the pro-inflammatory cytokines (tumor necrosis factor-alpha, interleukin-6) GSH level, kidney, and liver functions as proved by biochemical histopathological and immunohistochemical findings. This research introduces a novel effective protection modality for the γ-irradiation hazard via biocompatible CUR/HA NC injection.

In silico study on probing atomistic insights into structural stability and tensile properties of Fe-doped hydroxyapatite single crystals

Hydroxyapatite (HA, Ca10PO4(OH)2) is a widely explored material in the experimental domain of biomaterials science, because of its resemblance with natural bone minerals. Specifically, in the bioceramic community, HA doped with multivalent cations (e.g., Mg2+, Fe2+, Sr2+, etc.) has been extensively investigated in the last few decades. Experimental research largely established the critical role of dopant content on mechanical and biocompatibility properties. The plethora of experimental measurements of mechanical response on doped HA is based on compression or indentation testing of polycrystalline materials. Such measurements, and more importantly the computational predictions of mechanical properties of single crystalline (doped) HA are scarce. On that premise, the present study aims to build atomistic models of Fe2+-doped HA with varying Fe content (10, 20, 30, and 40 mol%) and to explore their uniaxial tensile response, by means of molecular dynamics (MD) simulation. In the equilibrated unit cell structures, Ca(1) sites were found to be energetically favourable for Fe2+ substitution. The local distribution of Fe2+ ions significantly affects the atomic partial charge distribution and chemical symmetry surrounding the functional groups, and such signatures are found in the MD analyzed IR spectra. The significant decrease in the intensity of the IR bands found in the Fe-doped HA together with band splitting, because of the symmetry changes in the crystal structure. Another important objective of this work is to computationally predict the mechanical response of doped HA in their single crystal format. An interesting observation is that the elastic anisotropy of undoped HA was not compromised with Fe-doping. Tensile strength (TS) is systematically reduced in doped HA with Fe2+ dopant content and a decrease in TS with temperature can be attributed to the increased thermal agitation of atoms at elevated temperatures. The physics of the tensile response was rationalized in terms of the strain dependent changes in covalent/ionic bond framework (Ca–P distance, P–O bond strain, O–P–O angular strain, O–H bond distance). Further, the dynamic changes in covalent bond network were energetically analyzed by calculating the changes in O–H and P–O bond vibrational energy. Summarizing, the current work establishes our foundational understanding of the atomistic phenomena involved in the structural stability and tensile response of Fe-doped HA single crystals.

Porous ceramic coating formed on 316L by laser cladding combined plasma electrolytic oxidation for biomedical application

In order to improve the bioactivity of 316L stainless steel, a titanium layer was prepared on the surface of 316L by laser cladding (LC), followed by plasma electrolytic oxidation (PEO) to form a porous ceramic coating on titanium layer. The morphologies, microstructure and compositions of the coated samples were characterized by 3D surface profiler, SEM, EDS, XRD and XPS. The corrosion resistance and bioactivity of the coatings were evaluated by potentiodynamic polarization and immersion test in simulated body fluid (SBF), respectively. The results showed that the porous ceramic coating mainly consisted of anatase and rutile, and highly crystalline HA was also detected. The main elements of the PEO coating are Ca, P, Ti and O. The LC+PEO composite bio-coating has more excellent corrosion resistance than the 316L substrate in simulated body fluid. Furthermore, the composite coating could effectively improve the bioactivity of 316L stainless steel.

Electrochemical and In Vitro Biological Evaluation of Bio-Active Coatings Deposited by Magnetron Sputtering onto Biocompatible Mg-0.8Ca Alloy.

The use of resorbable magnesium alloys in the design of implants represents a new direction in the healthcare domain. Two main research avenues are currently explored for developing or improving metallic biomaterials: (i) increase of their corrosion resistance by designed compositional and structural modifications, and (ii) functionalization of their surfaces by coating with ceramic or polymeric layers. The main objective of this work was to comparatively assess bio-functional coatings (i.e., highly-crystallized hydroxyapatite and silica-rich glass) deposited by radio-frequency magnetron sputtering (RF-MS) on a biodegradable Mg-0.8Ca alloy (0.8 wt.% of Ca). After probing their morphology (by scanning electron microscopy) and structure (by Fourier transform infrared spectroscopy and grazing incidence X-ray diffraction), the corrosion resistance of the RF-MS coated Mg-0.8Ca substrates was electrochemically tested (in synthetic biological media with different degrees of biomimicry), and their cytocompatibility was assessed in osteoblast and fibroblast cell cultures. By collective assessment, the most promising performances, in terms of mass loss (~7% after 12 days), hydrogen release rate (~6 mL/cm2 after 12 days), electrochemical corrosion parameters and cytocompatibility, were obtained for the crystalline HA coating.

A potential trackable bone filler: Preparation and characterization

AbstractThis study reported the synthesis of fluorescent hydroxyapatite/alginate/carbon quantum dots (HA/Alg/CQDs) nanocomposites via the co‐precipitation technique. The N‐doped CQDs as a new class of fluorescent materials were prepared by the citric acid pyrolysis method, with an average size around 4 nm. Physical, chemical, and optical properties of the synthesized nanocomposites were investigated by X‐ray diffraction (XRD), Fourier‐transformed infrared spectroscopy (FTIR), atomic force microscopy (AFM), field‐emission scanning electron microscopy (FESEM), UV–visible spectroscopy, and photoluminescence (PL) spectroscopy, respectively. The PL spectroscopy data verified the favorable in vitro luminescent emission of the HA/Alg/CQDs nanocomposites in comparison with HA/Alg and HA samples. The XRD patterns of the prepared samples confirmed the formation of crystalline HA in all composites, possessing a Ca/P ratio around 1.5 as obtained by EDX elemental analysis. The FESEM analysis exhibited HA nanoplates that homogeneously distributed throughout the alginate matrix. Therefore, the synthesized nanocomposites could be regarded as potential trackable drug carriers for hard tissue engineering applications.

Probiotic bacteria cell surface-associated protein mineralized hydroxyapatite incorporated in porous scaffold: In vitro evaluation for bone cell growth and differentiation

There is a high demand for synthesis of biocompatible hydroxyapatite nanoparticle (HANP), which is a key component in bone tissue engineering scaffolds. The present study describes a facile route of HANP synthesis through mineralization of the cell surface-associated protein (CSP) from the human probiotic lactic acid bacteria (LAB) Lactobacillus rhamnosus GG. CSP extract from the LAB (consisting of ~66 kDa, ~47 kDa, ~40 kDa and ~25 kDa protein) was mineralized to yield spindle-shaped HANPs having an average particle length of 371 nm as evidenced in FETEM analysis. CSP-mineralized HANPs (CSP-HANPs) were characterized by FTIR and BET analysis, while XRD and SAED analysis indicated their crystalline nature. Mechanistic studies suggested the key role of ~25 kDa CSP (F4SP) in mineralization. In contrast to CSP-HANPs, F4SP-mineralized crystalline HA was plate-shaped having an average length of 1.68 μm and breadth of 0.95 μm. HANP mineralization at the whole-cell (WC) level resulted in clusters of aggregated HANPs (WC-HANPs) adhering onto L. rhamnosus GG cells as evident in FETEM, FESEM and AFM analysis. FETEM analysis revealed that the desorbed WC-HANPs recovered by cell lysis were needle-shaped, with a particle size distribution of 70–110 nm. Given that CSP-HANPs were non-toxic to cultured HEK 293 cells and osteoblast-like MG-63 cells, chitosan-gelatin (CG) scaffold incorporated with 15% w/v CSP-HANP (H-CG) was generated and tested for bone cell growth. H-CG exhibited a favorable pore size distribution (160–230 μm), overall porosity (~84%) and biodegradation profile. H-CG scaffold was conducive to osteogenesis and rendered enhanced proliferation, alkaline phosphatase (ALP) activity, calcium mineralization and heightened marker gene expression (ALP, Col I, Runx2 and OCN) in seeded MG-63 cells. CSP sourced from a safe probiotic LAB is thus a viable and effective mineralization template for synthesis of biocompatible HANPs that can be leveraged for bone tissue engineering applications.

Silver-Doped Hydroxyapatite Coatings Deposited by Suspension Plasma Spraying

Pure hydroxyapatite suspension was produced by wet chemical synthesis. Using a hybrid water-stabilized torch, a series of HA coatings were produced on SS304 and Ti6Al4V substrates and their properties were characterized by SEM, EDX and XRD techniques. After deposition, the amorphous phase content reached 6-10% and the coatings retained 75-82% of crystalline HA phase. Their thickness reached 145 μm. To understand the wear behavior of the coatings, pin-on-disc tribology evaluation was performed. Additionally, a set of HA coatings was prepared with pure metallic Ag content. This formed by in situ chemical decomposition of AgNO3 added into the HA suspension. The Ag was dispersed evenly within the coatings in the form of submicron-sized particles situated predominantly along the HA splats boundaries with a total Ag content of 8 wt.%. Given the antibacterial properties of Ag, such result presents a promising step forward in the hard tissue replacement research.

Mechanosynthesis of hydroxyapatite–ferrite composite nanopowder

In the present work, an investigation of the mechanosynthesis of calcium hydroxyapatite (HA, Ca10(PO4)6(OH)2) from a mixture of calcium oxide (СаО) and ammonium hydrophosphate ((NH4)2HPO4) and mechanotreatment of HA in a planetary mill with the use of steel drums and milling body has been performed. The obtained results have shown that the mechanosynthesis of crystalline nanodisperse HA proceeds through the stage of formation of an amorphous material. The temperature treatment of HA powders at 1000°C has enabled us to establish the influence of the treatment time on the phase composition of the powders and establish the following sequence of phase transformations: Ca10(PO4)6(OH)2→β-Ca3(PO4)2 (tmilling~2h), β-Ca3(PO4)2→α-Ca3(PO4)2 (tmilling~5h), β-,α-Ca3(PO4)2→Ca10(PO4)6(OH)2 (tmilling~7h).The mechanosynthesis and mechanotreatment of hydroxyapatite in steel drums with steel balls is accompanied by the contamination of hydroxyapatite by their wear debris (iron + manganese). A large part of oxidized iron forms superparamagnetic inclusions distributed in HA powder. A small part of Fe3+ and Mn2+ ions from the steel wear debris enters into the hydroxyapatite lattice, substituting Ca2+ ions. As a result, a nanocomposite powder consisting of hydroxyapatite, alloyed by Fe3+ and Mn2+ ions and ferrite inclusions forms. The phase composition of HA powders, the degree of their alloying by Fe3+ and Mn2+ ions, and the content of ferrite inclusions can be controlled by changing the time of mechanotreatment.

HA/BG composite films deposited by pulse laser under O2 atmosphere

HA/BG composite films have been deposited on Ti-6Al-4V by pulse laser under O2 atmosphere, and EPMA-EDS, SEM, XRD, FTIR measurements have been performed. Under O2 atmosphere, the crystallinity of the deposited films increases and Ca/P atomic ratio slightly decreases with increasing the substrate temperature. When the film was deposited at 600°C, the film was compact and crystalline HA was observed. Comparing with Ar atmosphere, O2 atmosphere is benefical for decreasing Ca/P atomic ratio of the films, and the in vivo test indicates that the film obtained under O2 atmosphere has excellent osteoinductivity. So O2 atmosphere is a promise alternative atmosphere to fabricate HA/BG composite films by pulsed laser deposition.

Hydroxyapatite coating with strong (002) crystallographic texture deposited by micro-plasma spraying

Micro-plasma spraying was used to fabricate HA coatings on Ti-6Al-4V substrate. The coating phase distribution significantly changed throughout the coating depth. Crystalline HA, ACP and ß-TCP were observed near the coating-substrate interface. However, HA content increased with increasing distance from interface, and it was ~90% at top surface of the coating (>40μm) according to micro-Raman analysis. Columnar structure was found in such region, which was related to a strong (002) crystallographic texture. In addition, dissolution behavior of the coating was evaluated by immersing the material in balanced salt solution for 14 days. The study revealed that HA coating exhibited superior chemical stability, and the columnar structure emerged on the surface of the coating during immersion experiment.

In situ synthesis of hydroxyapatite-grafted titanium nanotube composite

ABSTRACTThe present study is an investigation to demonstrate the effectiveness of in situ approach in the synthesis of hydroxyapatite-grafted titanium nanotube composite (HA-TNT). This method involves combining the process of HA sol–gel and rapid breakdown anodisation of titanium in a novel solution consisting of NaCl and N3PO4. This new synthesis approach produced a uniform dispersion of Anatase and Rutile phases of TiO2 nanotubes with minimal agglomeration in the matrix of crystalline HA. The characterisation of homogenised HA-TNT composite was investigated via field emission scanning electron microscopy (FESEM), energy dispersive spectroscopy (EDS), transmission electron microscope (TEM) and X-ray diffraction (XRD). FESEM and TEM images indicated the nanostructure of composite with TiO2 nanotube diameter of approximately 10 nm. XRD and EDS analyses confirmed the formation of HA crystalline with the Ca/P ratio of 1.58 and formation of Anatase and Rutile phase of TiO2 nanotubes.

Infusion of Ag‐Ion from Aqueous Solution into Solid Calcium Phosphate of Hydroxyapatite (HA) Crystal Structure

In contrast to extensive literature concerning Ag incorporation in hydroxyapatite, HA, while the phosphate approximated to stoichiometry of Ca10(PO4)6(OH)2, with added Ag has been precipitating from an aqueous solution, the paper presents Ag incorporation through Ag ion infusion from AgNO3 solution into solid HA pressed in pellet and ignited at 800°C. After Ag ions infused into the HA‐solid (crossed the interfacial solution‐solid boundary), they diffused across the crystal structure to a depth of time‐dependent several mm. The path of Ag diffusion in the solid HA was recorded using SEM‐EDS point analyses of Ag, Ca, P, EDS‐linear analyses of those elements, and elemental mapping. Time‐dependent concentrations of Ag+, Ca2+, and PO43− in AgNO3 solutions were also analyzed. The appearance of Ag in the crystalline HA with simultaneous local depletion in Ca and phosphate recorded as P, observed by EDS with simultaneous appearance of Ca2+ and PO43− ions and a decrease in Ag+ concentration in AgNO3 solution led the authors to a conclusion that Ag+ for Ca2+ substitution supported by PO43− charge balancing in the crystalline HA was in process. The HA particles in the section of the pellet without Ag had a uniform shape and size approximated to 300–400 nm. SEM image of the HA solid section, where Ag ions appeared was characterized by irregular aggregates of smaller crystals with sporadically present large, shaped in prism blocks identified by the XRD as Ag3PO4.

Biomimetic composite scaffolds based on mineralization of hydroxyapatite on electrospun poly(ɛ-caprolactone)/nanocellulose fibers

A biomimetic nanocomposite scaffold with HA formation on the electrospun poly(ɛ-caprolactone) (PCL)/nanocellulose (NC) fibrous matrix was developed in this study. The electrospun PCL/NC fiber mat was built and then biomineralized by treatment in simulated body fluid (SBF). Using such a rapid and effective procedure, a continuous biomimetic crystalline HA layer could be successfully formed without the need of any additional chemical modification of the substrate surface. The results showed that the introduction of NC into composite fibers is an effective approach to induce the deposition of HA nucleus as well as to improve their distribution and growth of a crystalline HA layer on the fibrous scaffolds. The water contact angle (WCA) of the PCL/NC/HA scaffolds decreases with increasing NC content and mineralization time, resulting in the enhancement of their hydrophilicity. These results indicated that HA-mineralized on PCL/NC fiber can be prepared directly by simply using SBF immersion.

Biomimetic composite microspheres of collagen/chitosan/nano-hydroxyapatite: In-situ synthesis and characterization

The collagen/chitosan/hydroxyapatite (COL/CS/HA) composite microspheres with a good spherical form and a high dispersity were successfully obtained using an in-situ synthesis method. The FT-IR and XRD results revealed that the inorganic phase in the microspheres was crystalline HA containing carbonate ions. The morphology of the composite microspheres was dependent on the HA content, and a more desirable morphology was achieved when 20wt.% HA was contained. The composite microspheres exhibited a narrow particle distribution, most of which ranged from 5 to 10μm. In addition, the needle-like HA nano-particles were uniformly distributed in the composite microspheres, and their crystallinity and crystal size decreased with the HA content.

Crystalline ha coating on peek via chemical deposition

• Hydroxyapatite crystalline particles successfully chemically deposited on PEEK. • All process was done in room temperature and eliminates subsequent sintering. • Three dimensional porous treated layer were produced via this method. Polyether ether ketone (PEEK) has a similar elastic modulus to bone and can be a suitable alternative to metallic implants. However, PEEK is bioinert and does not integrate well with the surrounding tissues. The current commercial method for solving this problem is by coating PEEK substrates with calcium phosphates via plasma spraying. However, this method produces a low bonding strength between the substrate and the coating layer, as well as non-uniform density of the coating. In this study, chemical deposition was used to deposit HA crystalline particles on PEEK substrate without any subsequent crystallisation process therefore producing crystalline treated layer. EDX results confirmed the deposition of HA, and the XRD results confirmed that the treated layer was crystalline HA. FT-IR analysis confirmed the chemical bonding between HA and the substrate. Surface roughness increased from 24.27 nm to 34.08 nm for 3 min immersion time. The water contact angle showed an increase in wettability of the treated sample from 71.6 to 36.4 degrees, which in turn increased its bioactivity. The proposed method is a suitable alternative to other conventional methods as high temperature was not involved in the process which could damage the surface of the substrate.

Chemical and Thermal Stability of Multiple Ions Doped Non-Stoichiometric Nanoapatite Heat-Treated in CO<sub>2</sub> and Air Atmospheres

Nanostructured apatite has been widely used as a bone substitute material due to its close resemblance to human bone mineral. To further mimic biological apatites, multiple ions doped non-stoichiometric nanoapatite has been studied. A nanosized apatite (NAp-2) containing Mg (1.09 wt%), Na (0.15 wt%), K (0.008 wt%) and CO32- (5.18 wt%) was synthesized by a wet precipitation technique. The presence of these ions in NAp-2 was detected using ICP. Broad diffraction peaks of XRD results indicated the presence of nanocrystalline phase pure NAp-2. The primary particle size of the resulted powder was ~ 20 nm, typical of bone crystal size, estimated using Scherrers equation. Based on CHN results, the NAp-2 powders showed a total loss of 51 and 78% of carbonate ions when heat-treated at 900°C in both CO2 and air atmospheres, respectively. This indicates that the heat-treatment in CO2 flux has reduced the carbonate ions lost from the NAp-2. A highly crystalline HA phase was formed in the ionic doped NAp-2 without secondary phases, indicating a thermal stability of this powder at 900°C in CO2 and air atmospheres. Thus, this study demonstrated that a phase pure multiple ions doped nanoapatite was synthesized using a wet precipitation technique.

Characterization of Thermal Sprayed Hydroxyapatite Coatings on Some Biomedical Implant Materials

To characterize two thermal sprayed coatings of different particle-sized hydroxyapatite on some biomedical implant materials. Characterization of the as-sprayed coatings was performed using x-ray diffractometer (XRD), scanning electron microscopy/energy dispersive spectroscopy (SEM/EDS), and x-ray mapping techniques. Bond strength of the sprayed coatings measured as per ASTM-C633 standard and cell culture studies were performed to access the biocompatibility of the coatings. The results indicated that, HA coatings of 10 μm particle sized powder were completely amorphous, while that of 30 μm sized powder consisted of crystalline HA with minor tetracalcium phosphate (TTCP) and tricalcium phosphate (TCP) phases. This flame spray HA-A and HA-B coatings exhibited higher bond strength (67.8 and 49.3 MPa respectively) in comparison with other thermal spray HA coating techniques reported in literature. In-vitro biocompatibility studies revealed that HA-B coated specimens had good biocompatibility with a human osteosarcoma cell line KHOSNP (R-970-5). These observations show that particle size of HA powders have a significant effect on the phase composition, microstructure, roughness and biocompatibility of deposited coatings. HA-B coatings were found to be metallurgically better than the HA-A coating from the point of view of biomedical applications.

The influence of new wet synthesis route on the morphology, crystallinity and thermal stability of multiple ions doped nanoapatite

Abstract The synthesis of multiple ions doped nanoapatite powder was carried out by wet precipitation technique. A newly developed reaction route with self-controlled pH at high reaction temperatures, i.e. 37 & 85±2 °C was compared to the conventional synthesis route at 37±2 °C. The XRD peaks were very broad, indicating the presence of nanocrystalline apatite. The primary particle size of the powder was in the range of 20–30 nm whereas the fraction of crystallinity was between 0.20 and 0.63. TEM and SEM characterizations confirmed the nanosized primary particles of the apatite samples. The high temperature synthesis at 37 & 85±2 °C improved both crystallite size and crystallinity of the as-prepared samples. A highly crystalline HA phase was formed in the ions doped samples without secondary phases, indicating its thermal stability at 900 °C in both CO2 and air atmosphere. The in vitro cytocompatibility of the synthesised nanoapatite powders was confirmed by cell viability of human skin fibroblasts.