In vitro measurement of the chemical changes occurring within ß-tricalcium phosphate bone graft substitutes.

Several mechanisms proposed to explain the osteoinductive potential of calcium phosphates involve surface mineralization ("bioactivity") and mention the occurrence of concentration gradients between the inner and the outer part of the implanted material. Determining the evolution of the local chemical environment occurring inside the pores of an implanted bone graft substitute (BGS) is therefore highly relevant. A quantitative and fast method was developed to measure the chemical changes occurring within the pores of ß-Tricalcium Phosphate (ß-TCP) granules incubated in a simulated body fluid. A factorial design of experiment was used to test the effect of particle size, specific surface area, microporosity, and purity of the ß-TCP granules. Large pH, calcium and phosphate concentration changes were observed inside the BGS and lasted for several days. The kinetics and magnitude of these changes (up to 2 pH units) largely depended on the processing and properties of the granules. Interestingly, processing parameters that increased the kinetics and magnitude of the local chemical changes are parameters considered to favor calcium phosphate osteoinduction, suggesting that the model might be useful to predict the osteoinductive potential of BGSs. STATEMENT OF SIGNIFICANCE: Recent results suggest that in situ mineralization of biomaterials (polymers, ceramics, metals) might be key in their ability to trigger ectopic bone formation. This is the reason why the effect on in situ mineralization of various synthesis parameters of ß-tricalcium phosphate granules was studied (size, microporosity, specific surface area, and Ca/P molar ratio). To the best of our knowledge, this is the first article devoted to the chemical changes occurring within the pores of a bone graft substitute. We believe that the manuscript will prove to be highly important in the design and mechanistic understanding of drug-free osteoinductive biomaterials.

Profex: a graphical user interface for the Rietveld refinement program BGMN.

Profex is a graphical user interface for the Rietveld refinement program BGMN. Its interface focuses on preserving BGMN's powerful and flexible scripting features by giving direct access to BGMN input files. Very efficient workflows for single or batch refinements are achieved by managing refinement control files and structure files, by providing dialogues and shortcuts for many operations, by performing operations in the background, and by providing import filters for CIF and XML crystal structure files. Refinement results can be easily exported for further processing. State-of-the-art graphical export of diffraction patterns to pixel and vector graphics formats allows the creation of publication-quality graphs with minimum effort. Profex reads and converts a variety of proprietary raw data formats and is thus largely instrument independent. Profex and BGMN are available under an open-source license for Windows, Linux and OS X operating systems.

Textured and hierarchically structured calcium phosphate ceramic blocks through hydrothermal treatment.

Synthetic calcium phosphate bone graft substitutes are widely recognized for their biocompatibility and resorption characteristics in the treatment of large bone defects. However, due to their inherent brittleness, applications in load-bearing situations always require reinforcement by additional metallic implants. Improved mechanical stability would eliminate the need for non-resorbable metallic implants. In this context a new approach to obtain calcium phosphate scaffolds with improved mechanical stability by texturing the material in specific crystal orientations was evaluated. Texture and reduction of crystal size was achieved by recrystallizing α-TCP blocks into calcium deficient hydroxyapatite (CDHA) under hydrothermal conditions. SEM and XRD analysis revealed the formation of fine CDHA needles (diameter ≈ 0.1-0.5 µm), aligned over several hundreds of micrometers. The obtained microstructures were remarkably similar to the microstructures of the prismatic layer of mollusk shells or enamel, also showing organization at 5 hierarchical structure levels. Brazilian disc tests were used to determine the diametral tensile strength, σdts, and the work-of-fracture, WOF, of the textured materials. Hydrothermal incubation significantly increased σdts and WOF of the ceramic blocks as compared to sintered blocks. These improvements were attributed to the fine and entangled crystal structure obtained after incubation, which reduces the size of strength-determining critical defects and also leads to tortuous crack propagation. Rupture surfaces revealed intergranular tortuous crack paths, which dissipate much more energy than transgranular cracks as observed in the sintered samples. Hence, the refined and textured microstructure achieved through the proposed processing route is an effective way to improve the strength and particularly the toughness of calcium phosphate-based ceramics.

Calorimetry investigations of milled α-tricalcium phosphate (α-TCP) powders to determine the formation enthalpies of α-TCP and X-ray amorphous tricalcium phosphate.

One α-tricalcium phosphate (α-TCP) powder was either calcined at 500°C to obtain fully crystalline α-TCP or milled for different durations to obtain α-TCP powders containing various amounts of X-ray amorphous tricalcium phosphate (ATCP). These powders containing between 0 and 71wt.% ATCP and up to 2.0±0.1wt.% ß-TCP as minor phase were then hydrated in 0.1M Na2HPO4 aqueous solution and the resulting heat flows were measured by isothermal calorimetry. Additionally, the evolution of the phase composition during hydration was determined by in situ XRD combined with the G-factor method, an external standard method which facilitates the indirect quantification of amorphous phases. Maximum ATCP hydration was reached after about 1h, while that of crystalline α-TCP hydration occurred between 4 and 11h, depending on the ATCP content. An enthalpy of formation of -4065±6kJ/mol (T=23°C) was calculated for ATCP (Ca3(PO4)2), while for crystalline α-TCP (α-Ca3(PO4)2) a value of -4113±6kJ/mol (T=23°C) was determined.

Effect of amorphous phases during the hydraulic conversion of α-TCP into calcium-deficient hydroxyapatite.

Powders of α-tricalcium phosphate (α-TCP), which readily react with water to form calcium-deficient hydroxyapatite (CDHA), are frequently used in bone cements. As, for clinical applications, it is important to adjust the setting reaction of the cements to a reasonable reaction time, exact knowledge of the hydration mechanism is essential. It is known that prolonged milling results in partial amorphization of α-TCP powders and that dissolution of the amorphous phase significantly accelerates the hydration, but it is not clear yet when the amorphous phase reacts in comparison to the crystalline α-TCP. Therefore the aim of this study was to investigate the development of quantitative phase content of α-TCP samples during hydration. For this purpose, three α-TCP powders, containing 0, 16 and 71wt.% of amorphous phase (ATCP), were mixed with either deionized water or a 0.1M Na2HPO4 aqueous solution. The crystalline evolution of the paste was assessed quantitatively during the first 48h of hydration at 23°C by G-factor quantification. The present investigations demonstrate that ATCP reacted earlier than crystalline α-TCP. The results also suggest the formation of an X-ray amorphous phase during the hydraulic conversion formation of α-TCP into CDHA.

Growth kinetics of hexagonal sub-micrometric ß-tricalcium phosphate particles in ethylene glycol.

Recently, uniform, non-agglomerated, hexagonal ß-tricalcium phosphate (ß-TCP) platelets (diameter≈400-1700nm, h≈100-200nm) were obtained at fairly moderate temperatures (90-170°C) by precipitation in ethylene glycol. Unfortunately, the platelet aspect ratios (diameter/thickness) obtained in the latter study were too small to optimize the strength of polymer-ß-TCP composites. Therefore, the aim of the present study was to investigate ß-TCP platelet crystallization kinetics, and based on this, to find ways to better control the ß-TCP aspect ratio. For that purpose, precipitations were performed at different temperatures (90-170°C) and precursor concentrations (4, 16 and 32mM). Solution aliquots were retrieved at regular intervals (10s-24h), and the size of the particles was measured on scanning electron microscopy images, hence allowing the determination of the particle growth rates. The ß-TCP platelets were observed to nucleate and grow very rapidly. For example, the first crystals were observed after 30s at 150°C, and crystallization was complete within 2min. The crystal growth curves could be well-fitted with both diffusion- and reaction-controlled equations, but the high activation energies (∼100kJmol(-1)) pointed towards a reaction-controlled mechanism. The results revealed that the best way to increase the diameter and aspect ratio of the platelets was to increase the precursor concentration. Aspect ratios as high as 14 were obtained, but the synthesis of such particles was always associated with the presence of large fractions of monetite impurities.

Control of the size, shape and composition of highly uniform, non-agglomerated, sub-micrometer ß-tricalcium phosphate and dicalcium phosphate platelets.

Calcium phosphates (CaPs) are widely used as bone graft substitutes but are inherently brittle, hence restricting their use to mechanically protected environments. Combining them with a tough polymer matrix could potentially lead to a composite with load-bearing properties. However, the highest mechanical properties can only be achieved if the CaP particles possess very precise features: they should be uniform in size and shape, non-agglomerated, elongated and thin. The aim of the present study therefore was to assess a novel method to produce such particles. This involved the precipitation of CaP particles in ethylene glycol at moderate temperatures (90-170 °C) and the variation of different reaction parameters (temperature, concentration, pH, etc) to study their influence on particle composition, size, shape and dispersion was studied. As a result, two main CaP phases were obtained as well-dispersed and highly uniform platelets in the form of: (i) ß-tricalcium phosphate (ß-TCP) hexagonal prisms and (ii) monetite (DCP) flat parallelepipeds. The size dispersion was the narrowest for ß-TCP (standard deviation/mean < 5%) whereas the aspect ratio was the highest for DCP (up to 25). In both cases, the thickness of the platelets was below 300 nm which should be ideal for the synthesis of strong CaP-based composites.

Printability of calcium phosphate powders for three-dimensional printing of tissue engineering scaffolds.

Three-dimensional printing (3DP) is a versatile method to produce scaffolds for tissue engineering. In 3DP the solid is created by the reaction of a liquid selectively sprayed onto a powder bed. Despite the importance of the powder properties, there has to date been a relatively poor understanding of the relation between the powder properties and the printing outcome. This article aims at improving this understanding by looking at the link between key powder parameters (particle size, flowability, roughness, wettability) and printing accuracy. These powder parameters are determined as key factors with a predictive value for the final 3DP outcome. Promising results can be expected for mean particle size in the range of 20-35 µm, compaction rate in the range of 1.3-1.4, flowability in the range of 5-7 and powder bed surface roughness of 10-25 µm. Finally, possible steps and strategies in pushing the physical limits concerning improved quality in 3DP are addressed and discussed.

A physical approach to modify the hydraulic reactivity of alpha-tricalcium phosphate powder.

A microsized alpha-tricalcium phosphate (alpha-TCP) powder was calcined at various temperatures (350 degrees C