The preparation and characterization of HA/β-TCP biphasic ceramics from fish bones

The aim of this research is to observe the physicochemical characterization and evaluate the biocompatibility of the HA/β-TCP biphasic calcium phosphate ceramics (BCP) produced from fish bones. In addition, the mechanism of the formation of BCP after calcination of fish bones was discussed. Three kinds of fish bones (Salmo salar, Anoplopoma fimbria and Sardine) were prepared and calcined for one hour at different temperatures ranging from 600 °C to 1100 °C in a muffle furnace. The calcined bones were analyzed by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (GTA), inductively coupled plasma optical atom emissions spectroscopy (ICP-OES), X-ray fluorescence (XRF) and scanning electron microscopy (SEM). The in vitro cytotoxicity assessment was used to evaluate the biocompatibility of the biphasic ceramics. BCP materials were produced from all kinds of fish bones by calcination above 700 °C, the carbonated hydroxyapatite and multiple trace element were also found in the calcined bones. With the increase of temperature, the ratio of HA/β-TCP varied and the major organic components were progressively removed. The carbonated hydroxyapatite disappeared when temperature rises above 900 °C. Rising temperature also caused crystal growth that eventually gave rise to the increase of the BCP grain size and influenced the mesoporous structure. The BCP materials were confirmed to have no obvious cytotoxicity to mesenchymal stem cells (MSC) in the in vitro cytotoxicity assessment. Calcium-deficient hydroxyapatite(CDHA) may make up the major inorganic constituent of fish bones that could decompose to HA and β-TCP when calcined above 700 °C. 800–900 °C is considered to be the optimal temperature to fabricate BCP materials which contain more β-TCP, carbonated hydroxyapatite and retain distinct mesoporous structure while has good biocompatibility. With the unique composition and structure, these three kinds of fish-bone-derived BCP materials can be further applied to fabricate bioceramic scaffolds for biomedical applications.     

Reaction sintering of β-tricalcium phosphates and their mechanical properties

Various kinds of calcium oxides, carbonates and phosphates were used as the raw materials, and β-TCP ceramics was fabricated by reaction sintering at 1100 °C, and the sinterability, the reaction sintering behavior and mechanical properties of reaction-sintered β-TCP were investigated. Reaction-sintered bodies using CaHPO₄ + HAp consisted of single β-TCP phase, and bulk density and bending strength increased with extending sintering time. On the contrary, normal-sintered β-TCP synthesized using CaHPO₄ + HAp did not change in bulk density and bending strength with extending sintering time. Reaction-sintered body using CaHPO₄ + HAp as the raw materials showed higher bulk density and bending strength than normal sintered β-TCP.     

Sintering behavior and mechanical properties of hydroxyapatite ceramics prepared from Nile Tilapia (Oreochromis niloticus) bone and commercial powder for biomedical applications

In this work, Nile tilapia (Oreochromis niloticus) bone was calcined at 800 °C for 5 h in an air atmosphere to obtain hydroxyapatite powder (FB powder). The elemental composition, phase structure, and morphology of the FB powder were investigated and compared with commercial hydroxyapatite powder (SM powder). The FB-powder exhibited 1.01 at. % of Mg while the SM-powder showed Mg in ppm-level. Carbonate groups were detected in the two powders. Both HAp and β-tricalcium phosphate (β-TCP) structures were found in the FB powder, but the SM powder exhibit only the HAp phase. Irregular-shaped particles were observed in the FB powder. After the two HAp powders were sintered at 1200 °C and 1250 °C for 2 h (FB-1200, FB-1250, SM-1200, and SM-1250), the β-TCP intensity peaks of the FB-ceramic samples significantly increased with increasing sintering temperature. The highest relative density, well-packed grains, and β-TCP stabilization by Mg at the Ca5 site of the FB-1250 structure were the dominant factors governing the highest mechanical properties. Although high density was observed in the SM-1200 sample, Vickers hardness of the SM-1200 sample is lower than the FB-1250 sample. This may be attributed to the partial decomposition of HAp into β-TCP, α-tricalcium phosphate (α-TCP), and Ca₁₀(PO₄)₆O phases. In addition, the increase of grain size was the main factor that governs the increasing compressive strength and Young's modulus instead of density and phase decomposition of the SM-ceramic samples.     

Hydroxyapatite,tricalcium phosphate and biphasic materials prepared by a liquid mix technique

The Pechini based liquid-mix technique has been applied to prepare either single phases of hydroxyapatite –Ca₁₀(PO₄)₆OH₂– (OHAp), α and β-tricalcium phosphate –Ca₃(PO₄)₂–, (α-TCP, β-TCP) or biphasic calcium phosphates (BCP). Compositions with a Ca/P molar ratio between 1.5 and 1.667 were synthesized and subjected to a thermal treatment up to 1400 °C. α and β-TCP were both prepared from a Ca/P ratio of 1.5, but while β-TCP is isolated at 900 °C and remains stable up to 1100 °C, it is necessary to anneal at 1400 °C for 72 h to obtain pure α-TCP. OHAp is obtained as a single phase from a 1.667 Ca/P ratio after annealing at 1000 °C for 24 h and starts to decompose at 1400 °C. Between these two extremes a whole range of biphasic calcium phosphates can be prepared by using this technique with an accurate control of the starting reactants. These materials have been characterized by FTIR, XRF, BET, XRD and, based on this technique, a phase quantification determination (QXRD). The solubility of these products was tested in a buffered solution at 37 °C and pH=7.4.     

Enhanced in vitro inhibition of MCF-7 and magnetic properties of cobalt incorporated calcium phosphate (HAp and β-TCP) nanoparticles

The Co²⁺ (0.1 M) incorporated hydroxyapatite (HAp) and beta tricalcium phosphate (β-TCP) nanoparticles were synthesized by the microwave assisted technique and sintering of HAp respectively. The samples were thermally treated at temperatures ranging from 200 to 1000°C. The partial substitutions of Co²⁺ at the Ca²⁺ site of HAp were confirmed from the slight shift (～0.2°) in the (002) and (211) XRD peaks. The morphology of the nanoparticles was transformed from nanospheres to twinned particles on thermal treatment. In addition, the particle size of Co-600 was increased (from ～50 nm to ～100 nm) due to the recrystallization process. Further, the thermal treatment enhanced the crystallinity (41.15 to 90.16%), retentivity (M_r) and coercivity (H_c) of the nanoparticles. The cobalt incorporated HAp and β-TCP possessed paramagnetic property. The excellent bioactivity of β-TCP has been confirmed by the mineralization in simulated body fluid (SBF). Compared to HAp, β-TCP possessed better compatibility towards C2C12 cells on cobalt incorporation as evidenced by the in vitro cell viability. Moreover, both HAp and β-TCP have significantly inhibited the growth of MCF-7 on increasing the interaction time (72 h). Hence, the inhibition characteristics of Co²⁺ incorporated calcium phosphate (CaP) towards MCF-7 (without affecting the normal cells) demonstrate its competency as a potential material for cancer therapy over the already existing nanoparticles.     

Sintering behavior and growth mechanism of β-TCP in nanocrystalline hydroxyapatite synthesized by mechanical alloying

Nanocrystalline carbonated HAp powder has been synthesized successfully within 2 h by mechanical alloying the stoichiometric mixture of CaCO₃, CaHPO₄·2H₂O at room temperature under open air. To observe the sintering behavior of HAp the as-milled sample is sintered at different temperatures. The amorphous HAp phase (~14 vol%) in as-synthesized sample transforms completely to crystalline HAp after sintering at 700 °C and after sintering the sample at 800 °C, the crystalline HAp partially transforms to β-TCP phase. Presence of low content of β-TCP phase in HAp powder could be useful in artificial hard tissue applications. Increase in sintering temperature up to 1000 °C results in enhancement of decomposition rate of HAp into β-TCP phase. Microstructure characterization in terms of lattice imperfections and relative phase abundances in non-sintered and all sintered samples are made both by analyzing the respective XRD patterns using Rietveld's structure refinement method as well as TEM images. The growth mechanism of β-TCP from crystalline HAp phase has been proposed based on structure and microstructure characterizations of sintered samples.     

Fabrication and characterization of porous biphasic β-tricalcium phosphate/carbonate apatite alginate coated scaffolds

In this research, biphasic β-tricalcium phosphate/carbonate apatite (β-TCP/CO₃Ap) scaffolds incorporated with alginate were fabricated. Sodium alginate was extracted from local brown seaweed, Sargassum polycystum via calcium alginate process. Biphasic β-TCP/CO₃Ap scaffolds were fabricated by polymer reticulate method. β-TCP slurry was infiltrated into the polyurethane foam (PU) foam, then sintered up to 1300?°C, soaked for 4?h and immediately quenched in still air to form biphasic β-TCP/α-TCP scaffold. Biphasic β-TCP/α-TCP scaffold was then transformed to biphasic β-TCP/CO₃Ap scaffold by dissolution-precipitation reaction with 1?M of NaHCO₃ at 170?°C for 1, 3 and 5 days. Biphasic β-TCP/CO₃Ap scaffold from 5 days dissolution-precipitation reaction was chosen to incorporate with 1%, 3% and 5% of sodium alginate, respectively, as it has the highest composition of CO₃Ap phase. FTIR and FESEM analysis confirmed the presence of characteristic functional groups of sodium alginate. Mechanical strength of biphasic β-TCP/CO₃Ap scaffold improved by increasing the concentration of sodium alginate. The highest mechanical strength achieved was 26.38 kPa for biphasic β-TCP/CO₃Ap scaffold with 5% sodium alginate coating and it was chosen to further study with the addition of 1%, 3% and 5% microspheres. FESEM analysis confirmed the attachment of microspheres on the surface of alginate/biphasic β-TCP/CO₃Ap scaffold was successful.     

Electrical polarization and ionic conduction properties of β-tricalcium phosphate bioceramics with controlled vacancies by sodium ion substitution

With the aim to understand electric polarization mechanisms of β-tricalcium phosphate as an advanced biomaterial, Na ion-substituted β-Ca₃(PO₄)₂ (Na-β-TCPs) ceramics with controlled lattice vacancies were synthesized and structural refinement was performed by the Rietveld method. The Rietveld analysis revealed that Ca and vacancies at Ca(4) sites in the β-TCP structure decreased with an increase in Na substitution. Electrical measurements by the complex impedance method revealed that the conductivity and the activation energy calculated from Cole-Cole plots rapidly decreased to a constant value with an increase in Na substitution and decrease in vacancies. The thermally stimulated depolarization current (TSDC) curve of the electrically polarized Na-β-TCP showed one large peak at 530–610 °C. However, the accumulated charge decreased with an increase in Na ions and decrease in vacancies up to 2.37 mol%, after which it became constant. These results are consistent with the presumed formation of a dipole moment between aligned Ca²⁺ ions and their vacancies along the direction of the external polarization field applied at high temperature. We conclude that the large amount of stored charge in β-TCP caused by electrical polarization is due to the low site occupancy of calcium ions and vacancies at Ca(4) sites in the β-TCP structure, which is not the case for hydroxyapatite (HAp), as previously reported.     

Synthesis and properties of Zn and Zn–Mg-doped tricalcium phosphates obtained by Spark Plasma Sintering

β-tricalcium phosphate (Ca₃(PO₄)₂ or TCP) are essential biomaterials because of the chemical composition, high biocompatibility and osseointegration. However, their limited mechanical properties restrict their use to areas where high mechanical performances are not required. Spark Plasma Sintering (SPS) was selected out of the unconventional sintering methods in order to obtain high-density doped-TCP bioceramic materials. The main advantages of SPS are a high heating rate, low sintering temperatures and short residence times, producing bioceramics with full density and fine-grain microstructure. The main purpose was to design, obtain by SPS and characterize undoped β-TCP, 1ZnO-doped β-TCP and 1ZnO-1MgO codoped β-TCP (wt. %) bioceramics. All the obtained samples were visually semitransparent and mainly β-TCP was detected by X-ray analysis. Densification behavior was determined by Archimedes' method and microstructural features of the sintered specimens were analyzed by Field Emission Scanning Electron Microscopy (FE-SEM-EDX). The undoped and doped β-TCP bioceramics were mechanically characterized, specifically the modulus of elasticity and Vickers microhardness. The results are compared with equivalent samples obtained by conventional solid-state sintering (CS) reaction. A first study of biological behavior was carried out, specifically direct cell adhesion of MG-63 human osteoblast-like cells on the polished surfaces of β-TCP, 1ZnO-β-TCP and 1ZnO–1MgO-β-TCP dense samples were determined. The present study concludes that the SPS process together with the doping effect enhanced sinterability, mechanical and biological properties of Zn-TCP and Zn–Mg-TCP based materials.     

Influence of microstructure and phase composition on the nanoindentation characterization of bioceramic materials based on hydroxyapatite

The aim of the study was to investigate the influence of microstructure and phase composition on the mechanical behaviour of hydroxyapatite (HAp) and biphasic HAp/β-tricalcium phosphate (β-TCP) bioceramic materials using nanoindentation. The formation of β-TCP phase in the HAp ceramic had the predominant influence on the nanomechanical properties of compact ceramics. For investigated microstructures there appear to be a slight decrease in the elastic modulus with increasing load and a higher decrease in hardness, which are in agreement with upper bounds of the results reported in literature. Maximal value of reduced modulus and hardness is yielded with pure HAp, and is measured to be 133.76 GPa for average grain size of 3 μm and 12.18 GPa for average grain size of 140 nm, respectively. The average modulus and hardness results for HAp/β-TCP ceramics with higher (101.61 GPa, 6.76 GPa) and lower grain size (115.72 GPa, 8.76 GPa) show sufficient mechanical properties in order to serve as hard tissue replacement material.     

Formation of a new solid β tricalcium orthophosphate structure,from reaction between hydroxyapatite and ammonium sulfate

The reaction which occur during heating, from room temperature to 1100°C, of a mixture of hydroxyapatite, Ca₁₀(PO₄)₆(OH)₂ [HAP] and ammonium sulfate (NH₄)₂SO₄ [AS] are studied. The formation of Ca₂(NH₄)₂(SO₄)₃, Ca₂P₂O₇ and Ca(PO₃)₂ is observed between 200°C and 300°C; at 400°C CaSO₄ appears. From 500 to 700°C, Ca(PO₃)₂ reacts with Ca SO₄ and with HAP and gives β-Ca₂P₂O₇. Lastly, from 700°C to 1000°C, β-Ca₂P₂O₇ reacts with HAP and with CaSO₄ and gives β-Ca₃(PO₄)₂ [β-TCP]; From 1000 to 1100°C, β-TCP and CaSO₄ react and form a sulfate ion containing calcium phosphosulfate, the structure of which is β-TCP     

In vitro evaluation of natural hydroxyapatite from Osteoglossum bicirrhosum fish scales for biomedical application

This study reports the obtainment of bioactive hydroxyapatite (HAp) extracted from scales of arowana fish (FSHA) (Osteoglossum bicirrhosum) by alkaline treatment followed by calcination at 600 and 800°C. The cell viability and bioactivity of hydroxyapatite particles (FSHA) were investigated and compared with those of HAp synthesized (s.HA) by the precipitation method. The HAp particles from fish scales showed non-toxic behavior to dental pulp stem cells similar to HAp synthesized. Additionally, bioactivity assays show that the Hap from natural source forms the bone-like apatite layer faster than s.HA sample, after being incubated in McCoy medium for 3 days. The results illustrate that HAp obtained from Osteoglossum bicirrhosum fish scale bio-waste showed excellent biocompatibility. Besides, this study provides an effective method for converting low-cost bio-waste into a value-added and it can be a potential alternative biomaterial for various biomedical applications.     

Improvement of mechanical properties of macroporous β-tricalcium phosphate bioceramic scaffolds with uniform and interconnected pore structures

The biocompatible and degradable macroporous bioceramic scaffolds with high mechanical properties and interconnected porous structures play an important role in hard tissue regeneration and bone tissue engineering applications. In this study, the improvement of mechanical properties of macroporous β-tricalcium phosphate [β-Ca₃(PO₄)₂, β-TCP] bioceramic scaffolds with uniform macropore size and interconnected pores were fabricated by impregnation of the synthesized β-TCP nano-powder slurry into polymeric frames. The microstructures, mechanical properties and in vitro degradation of the fabricated samples were investigated. For a comparison, β-TCP scaffolds were also fabricated from commercial micro-size powders under the same conditions. The resultant scaffolds showed porosities ∼65% with uniform macropore size ranging from 400 to 550 μm and interconnected pore size ∼100 μm. The compressive strength of the samples fabricated from nano-size powders reached 10.87 MPa, which was almost twice as high as those fabricated from commercial micro-size powders, and was comparable to the high-end value (2–10 MPa) of human cancellous bone. Furthermore, the degradation of the β-TCP bioceramics fabricated from nano-size powders was apparently lower than those fabricated from commercial micro-size powders, suggesting the possible control of the degradation of the scaffolds by regulating initial powder size. Regarding the excellent mechanical properties and porous structures, the obtained macroporous β-TCP bioceramic scaffolds can be used in hard tissue regeneration and bone tissue engineering applications.     

In situ formation of biphasic calcium phosphates and their biological performance in vivo

The co-precipitation technique has been applied to synthesize biphasic calcium phosphate (BCP). After annealing at 900 °C for 24 h, hydroxyapatite (HAp) and β-tricalcium phosphate (β-TCP) were obtained as a single phase at 1.67 and 1.5 Ca/P ratios, respectively. Between these two extremes, a whole range of BCP preparations could be synthesized by using this technique with an accurate control of starting reactants. The biological performance of BCP granulates with a specific content of 62% HAp and 38% β-TCP was investigated. After immersion in Hanks’ balanced salt solution (HBSS) for 1 week, a precipitation started to be formed with individual small granules on the specimen surface. An MTT assay indicated that BCP granulates have no cytotoxic effects on MG-63 cells, and that they have good biocompatibility. An implantation experiment in mouse skulls revealed that BCP granulate provides a strong positive effect on bone formation in vivo in mice.     

Phase and melting relationships of β, α and α′-Ca3(PO4)2 polymorphs in the Ca3(PO4)2-Zn3(PO4)2 system

In order to provide an exact knowledge of the phase transitions and melting relationships of Ca₃(PO₄)₂ (TCP) in the presence of zinc, a revisited version of the rich-Ca₃(PO₄)₂ region of the phase diagram of the system Ca₃(PO₄)₂-Zn₃(PO₄)₂ has been established in the present work. Experimental determination of this diagram was carried out by solid-state reactions of samples prepared from pure NH₄H₂PO₄, CaCO₃ and ZnO raw materials. X-ray Diffraction, Differential Thermal Analyses and Field Emission Scanning Electron Microscopy studies allowed to revise the α, β, α + β-TCP phase stability fields, delimitating for the first time the biphasic α + α′-TCP field and the melting relationships in the high temperature region of the system. The results allowed to determine two peritectic invariant points, at ≈1400 °C for 95 mol% Ca₃(PO₄)₂ and at ≈1490 °C for ≈99.5 mol% Ca₃(PO₄)₂.     

Mg2+, Sr2+, Ag+, and Cu2+ co-doped β-tricalcium phosphate: Improved thermal stability and mechanical and biological properties

β-tricalcium phosphate (β-TCP, β-Ca₃(PO₄)₂) is an attractive biomaterial for bone repair applications. However, its sintering and mechanical properties are limited by a problematic phase transition to α-TCP. Cationic doping of β-TCP is able to postpone the formation of α-TCP allowing higher sintering temperatures and better mechanical properties. The co-doping of β-TCP with Mg²⁺ and Sr²⁺ has already been studied in detail, but the addition of antibacterial cations (Ag⁺ and Cu²⁺) on the Mg–Sr β-TCP co-doped composition remains unexplored. Thus, two co-doped β-TCP compositions were realized by aqueous precipitation technique without any secondary phase and compared with undoped β-TCP: Mg–Sr (2.0–2.0 mol%) and Mg–Sr–Ag–Cu (2.0–2.0–0.1–0.1 mol%). Differential thermal analysis and dilatometry analyses showed a slight decrease of the β-TCP → α-TCP phase transition temperature for the Mg–Sr–Ag–Cu (2.0–2.0–0.1–0.1% mol) composition as compared to the Mg–Sr (2.0–2.0 mol%). However, both exhibited much higher transition temperatures than undoped β-TCP. The addition of Ag⁺ and Cu²⁺ slightly reduces the grain size after sintering compared to the Mg–Sr (2.0–2.0 mol%) and the undoped compositions. The co-doped compositions also exhibited improved mechanical properties, specifically a higher Vickers hardness and elastic modulus. Finally, cell proliferation assays showed that the presence of dopants, even Ag⁺ and Cu²⁺, does not affect the survival and proliferation of cells. Thus, the use of Mg²⁺, Sr²⁺, Ag⁺, and Cu²⁺ co-doped β-TCP could be very promising for biomedical applications due to the improvements of these dopants on the thermal stability and mechanical and biological properties.     

Fabrication of doped β-tricalcium phosphate bioceramics by Direct Ink Writing for bone repair applications

β-tricalcium phosphate (β-TCP, β-Ca₃(PO₄)₂) is one of the most attractive biomaterials for bone regeneration and β-TCP macroporous scaffolds are very promising for both cell proliferation and mechanical support. The Additive Manufacturing (AM) process called Direct Ink Writing (DIW), based on the extrusion of a concentrated ceramic slurry, is particularly adapted to resolve the main drawbacks associated with conventional shaping of ceramic scaffolds. In this work, co-doped β-TCP powders were synthetized and used to print macroporous scaffolds by DIW. Doped β-TCP powders have been proved to exhibit higher thermal stability, densification and mechanical properties compared to undoped β-TCP. Two co-doped compositions were produced via the aqueous precipitation technique combining magnesium, strontium, silver and copper cations: Mg-Sr (2.0–2.0 mol%) and Mg-Sr-Ag-Cu (2.0–2.0–0.1–0.1 mol%). DIW slurries were optimized with undoped and co-doped β-TCP with the use of a dispersant and a carboxymethylcellulose and polyethyleneimine mixture to obtain aqueous slurries filled with 42 vol% of powder. Complete rheological characterizations were realized to assess the suitability of the β-TCP slurries for the DIW process (shear-thinning and thixotropic behaviour). The whole processing chain including printing, osmotic drying (PEG 10000) and sintering (1100 °C, 3 h) was optimized to successfully print co-doped β-TCP macroporous scaffolds. Characterizations after sintering showed a reduction of macropores and microcracks using co-doped β-TCP powders as well as improved compressive strengths and densities compared to undoped β-TCP. A significant enhancement of compressive strength values was obtained compared to literature data.     

医用双相磷酸钙陶瓷的制备工艺研究

医用双相磷酸钙（BCP）陶瓷是β-磷酸三钙（β-TCP）和羟基磷灰石（HA）的复合体,其成分与骨矿物组成类似。它具有良好的生物相容性,在生物医学领域具有非常广阔的应用前景。且在生理环境下能发生不同程度的降解,被组织吸收。通过化学沉淀法制备纳米羟基磷灰石,然后通过可溶性钙盐和磷酸盐反应工艺制得β-磷酸三钙,最后将二者进行机械复合而制得双相磷酸钙,将所得样品用X射线衍射仪（XRD）进行了表征。结果显示：所得的双相磷酸钙中掺杂有β-焦磷酸钙,但是它的结晶较好,并且可以改善双相磷酸钙陶瓷的力学性能。     

Rapid synthesis of cesium-doped hydroxyapatite nanorods: characterisation and microbial activity

Rapid synthesis of cesium-doped hydroxyapatite (Ca₁₀(PO₄)₆(OH)₂, (HAp)) by rapid microwave synthesis technique is reported. The crystal, chemical, and dielectric properties, along with the phase composition and morphology were investigated. SEM study proved successful synthesis of HAp nanorods. The dielectric properties were affected by the doping amount of Cs-ions in HAp. The values of crystallinity degree at different concentrations of Cs-ions were decreased on increasing the Cs content. The antimicrobial activities of Cs-doped HAp showed high effects of growth inhibition on Staphylococcus aureus, S. saprophyticus, Escherichia coli, Haemophilus influenza, Candida albicans, and C. tropicalis. The concentration of 0.32 Cs-doped HAp has the high inhibition effect against S. aureus with the percentage of inhibition to 88.89%. The microbial inhibition growth reflected on the MICs and MBCs and MFCs. This newly designed Cs-doped HAp can be applicable in wide scales biomedical applications such as bone cement engineering and antimicrobial agents.     

Counterions present in syntheses induce the precipitation of two different populations of Sr-containing hydroxyapatite crystals

The ions found in biological environment during biomineralization as well as the counterions present in syntheses, i.e., the reagents employed, play an important role in the precipitation and stabilization of apatites. In this work, precipitation of strontium-containing hydroxyapatites at different concentrations (0, 20, 40, 60, 80 and 100% Sr²⁺) was performed in the presence of two counterions typically present in biological environments: Na⁺ and Cl⁻. It was demonstrated that the presence of Na⁺/Cl⁻ in the synthesis environment led to the precipitation of a biphasic hydroxyapatite (HAp) system formed by two non-miscible phases: Ca-rich HAp and Sr-rich HAp. The biphasic system was observed for intermediate Sr²⁺ concentrations (20, 40 and 60%) and exhibited greater lattice microstrain compared with the single-phase systems of Ca HAp (0%) and Sr HAp (100%). Although Na⁺/Cl⁻ were inserted into both Ca- and Sr-rich HAp phases, Cl⁻ ions were preferentially accommodated into the enlarged Sr-rich HAp structure. The presence of Cl⁻ ions in the Sr-rich HAp phase decreased microstrain compared to the Ca-rich phase. After calcination, the biphasic system was transformed into a completely miscible Ca/Sr HAp phase without the formation of other phosphates or oxides.