Massimo Gandolfi

Nanocrystals of magnesium and fluoride substituted hydroxyapatite

Hydroxyapatite nanocrystals synthetized in the presence of different concentrations of magnesium and fluoride ions in solutions--1, 5 and 10 at.% have been submitted to a structural and chemical characterization. The syntheses were carried out in the presence of low molecular weight polyacrylic acid, which has been verified to inhibit hydroxyapatite crystallization. The polyelectrolyte is adsorbed into the crystals during the synthesis and provokes a reduction of the mean crystal sizes. The reduction is greater along the direction orthogonal to the c-axis, suggesting a preferential adsorption of the polyelectrolyte on the crystalline faces parallel to the c-axis. Both magnesium and fluoride can be incorporated into the hydroxyapatite structure. On the basis of the values of the lattice constants and of the magnesium relative content of the solid phase, it can be suggested that probably just a part of magnesium is substituted for calcium, the remainder being adsorbed on the crystal surface. However, magnesium destabilizes the apatitic structure favouring its thermal conversion into beta-tricalcium phosphate, and displays an inhibiting effect on the crystallization of hydroxyapatite. This last effect is enhanced by the simultaneous presence of polyacrylic acid. Fluoride substitution for hydroxyl ions into hydroxyapatite structure induces a slight increase of the crystal sizes along the c-axis direction. The data indicate that the experimental approach can be successfully used to prepare nanoapatite with crystallinity, crystal dimensions, composition, structure and stability very close to those characteristics of biological apatites.

Inhibiting effect of zinc on hydroxylapatite crystallization

Abstract X-Ray diffraction and spectrophotometric analysis have been used to investigate the role of zinc on hydroxylapatite (HA) crystallization. The presence of zinc in solution strongly inhibits the crystallization of hydroxylapatite, which can be synthesized as a unique crystalline phase only up to zinc concentration of about 25 atom %. This phase exhibits a reduction of Ca/P molar ratio and crystal sizes with increasing zinc concentration. Although the Ca/Zn ratio in the solid phase is almost equivalent to that in solution, the values of the cell parameters of the apatitic phase indicate that zinc cannot appreciably substitute for calcium in HA structure. Therefore, zinc must be assumed to be adsorbed on the surface of apatite crystallites and/or in the amorphous phase. The extent of thermal conversion of HA into s-tricalcium phosphate (s-TCP) increases with increasing zinc concentration in the solid phase, either when it is obtained by means of synthesis in solution or after cyclic pH fluctuation. The decrease of the lattice constants of s-tricalcium phosphate with increasing zinc concentration in the solid phase indicates that zinc partially replaces calcium in this structure. The inhibiting effect of zinc on HA crystallization and its preference for s-TCP structure closely resembles the behavior previously observed for magnesium.

Isomorphous substitutions in β-tricalcium phosphate: The different effects of zinc and strontium

Abstract X-ray diffraction and infrared absorption analyses have been carried out on zinc-substituted and strontium-substituted β-tricalcium phosphate prepared by solid-state reaction. Zinc can substitute calcium up to 20 atom %, inducing a nonlinear variation of the lattice constants and an increase in degeneracy of the PO 4 3− infrared absorption bands. On the other hand, up to 80 atom % of strontium can enter into the crystal structure of β-tricalcium phosphate, causing a linear enlargement of the unit cell, in agreement with its greater ionic radius compared to that of calcium. Furthermore, strontium incorporation provokes the shift of the PO 4 3− absorption bands toward lower frequencies. On the basis of the data previously obtained on magnesium-substituted β-tricalcium phosphate, the different behaviours exhibited by zinc and strontium could be attributed to a different distribution into the cationic sites of the β-tricalcium phosphate structure. The results allow us to relate the effect of bivalent ions on the structure and relative stability of calcium phosphates with their ionic radius, and can be utilized to interpret the role of ionic composition on the properties of biological phosphates.

Crystal to crystal transformations and polymorphism in anionic hydrogen bonding networks stabilized by crown ether metal complexes.

15-crown[5] or 18-crown[6] complexes of alkali, transition metal and ammonium cations together with polyprotic inorganic and organic anions have been used to construct crystalline molecular salts based on hydrogen bonded anionic networks. This new class of organic-inorganic complexes displays a variety of crystal-to-crystal transformations, mainly associated to the loss/uptake of water molecules and/or to the ionic reorganization accompanying phase transitions on varying the temperature. The dehydration and phase transition processes have been investigated by DSC, TGA and variable temperature X-ray powder diffraction. Most of the complexes described herein have been prepared by solid state mixing of the solid reactants.

Structural modifications of hydroxyapatite induced by lead substitution for calcium

Continuous series of solid solutions of lead and calcium hydroxyapatites have been prepared by solid-state reaction as well as from aqueous solutions and characterized by X-ray diffraction and IR spectroscopic analyses. While the lattice constant a of the solid solutions varies linearly with composition, the variation of the lattice constant c does not follow Vegards law but shows a discontinuity at about 50% lead atom content. A similar discontinuity has been observed in the variation of the IR frequencies with composition. Although lead incorporation seems to destabilize the apatite structure, the solid solutions prepared from aqueous media at a lead content around 50% exhibit a high thermal stability. The variation of c, thermal stability, as well as the frequencies and intensities of the IR bands of lead–calcium hydroxyapatite are interpreted on the basis of the inhomogeneous lead distribution in the two non-equivalent cation sites of the hydroxyapatite structure.

X-ray diffraction study of in vitro calcification of tendon collagen.

Decalcified samples of turkey leg tendon were submitted to in vitro calcification in the presence of metastable solutions of calcium phosphate at different concentrations. The structural relationship between apatitic deposits and collagen fibrils was examined by high- and small-angle X-ray diffraction using conventional and synchrotron radiation sources. At high supersaturation the apatitic crystallites were deposited on the collagen fibrils with their crystallographic c-axis preferentially oriented parallel to the fibril axis. At lower supersaturation, a fraction of the apatitic crystallites also grew with the c-axis preferentially oriented parallel to the collagen fibril axis, whereas other exhibited a preferential orientation perpendicular to the fibril axis. The analysis of the small-angle X-ray diffraction data indicates that the deposition of the apatitic phase in the sample stored in solution at lower supersaturation induced modifications of the collagen electron density distribution in the axial direction, which can be attributed to the deposition of the inorganic crystallites inside the gap region of the collagen structure.

Thin-layer chromatographic determination of indolic tryptophan metabolites in human urine using Sep-Pak C18 extraction

Tryptophan and some of its indole metabolites were separated by thin-layer chromatography, stained with the Van Urk--Salkowski reagent, and quantitated by scanning densitometry. The application of this technique for the detection of the indoles in urine samples, employing Sep-Pak C18 cartridges for extraction, was demonstrated. The proposed method is simple and accurate. The detection limits were 2 micrograms/ml 5-hydroxytryptophan, 1.75 micrograms/ml 5-hydroxyindolyl-3-acetic acid, 1.5 micrograms/ml tryptophan, 0.8 micrograms/ml indolyl-3-acetic acid, 0.9 micrograms/ml indolyl-3-butyric acid, 1.75 micrograms/ml serotonin, and 1.25 micrograms/ml tryptamine.

Spray-congealed solid lipid microparticles as a new tool for the controlled release of bisphosphonates from a calcium phosphate bone cement

Graphical abstract Figure. No Caption available. Abstract The aim of this work was to develop an innovative drug delivery system potentially useful for the local delivery of Bisphosphonates to bone tissue. We propose the use of Solid Lipid Microparticles (MPs), up to now mainly used for oral and topical drug delivery, as carrier for bisphosphonates due to the favourable biocompatibility and lower toxicity of the lipids compared with many polymers. The delivery platform consisted of a biomimetic &agr;‐tricalcium phosphate‐gelatin cement (CPC) enriched with alendronate loaded MPs (MPs‐AL) produced by the spray congealing technology. Alendronate direct addition to cement composition is limited since Alendronate is able to sequester calcium from calcium phosphates, thus preventing the setting of the cements. At variance, this approach permitted to load a relatively high amount of the drug on the CPC and allowed the controlled release of the highly water soluble alendronate. A Design of Experiment (DoE) was employed for the screening of the effects of the formulation variables related to the presence of unloaded microparticle (MPs) on the cement most important mechanical properties. Then, MPs loaded with 10% w/w of alendronate were produced using five different carriers (Stearic Acid, Stearilic Alcohol, Cutina HR, Tristearin and Precirol ATO5). All MPs‐AL exhibited a spherical shape, encapsulation efficiency higher than 90% and prevalent particle size ranging from 100 to 150 &mgr;m. Solid state characterization (DSC, HSM and X‐ray powder diffraction) demonstrated that encapsulation of alendronate into MPs did not alter its crystal structure. MPs‐AL addition to the cement provoked a modest lengthening of the setting times and of the hardening reaction leading to the complete transformation of &agr;‐tricalcium phosphate into calcium‐deficient hydroxyapatite, without significantly affect the cement mechanical properties. Moreover, the results of in vitro AL release study performed on cements enriched with MPs‐AL showed that the system allows a controlled release of the drug over time.