Synthetic carbonated apatite ceramics are considered as promising alternative to auto- and allograft materials for bone substitute. The aim of this study was to investigate the thermal stability of an AB-type carbonated apatite in the wide temperature range. The data on the thermal stability have to allow the conditions of the sintering of the ceramics to be controlled. Initial carbonated apatite powders were prepared by interaction between calcium oxide and ammonium hydrogen phosphate with addition of ammonium carbonate. Decomposition process was monitored by infra red spectroscopy, weight loss and X-ray diffraction of solid, and by infra red analysis of condensed gas phase resulted from the thermal decomposition of the sample in equilibrium conditions. Features of carbon monoxide and carbon dioxide release were revealed. The synthesized AB-type carbonated apatite is started to decompose at about 400 degrees Celsius releasing mainly carbon dioxide, but retained some carbonate groups and apatite structure at the temperature 1100 degrees Celsius useful to prepare porous carbonate-apatite ceramics intended for bone tissue engineering scaffolds.

A blue calcium phosphate cement with optimal self-hardening properties was synthesized by doping whitlockite (β-TCP) with copper ions. The mechanism and the kinetics of the cement solidification process were studied using energy dispersive X-ray diffraction and it was found out that hardening was accompanied by the phase transition from TCP to brushite. Reduced lattice parameters in all crystallographic directions resulting from the rather low (1:180) substitution rate of copper for calcium was consistent with the higher ionic radius of the latter. The lower cationic hydration resulting from the partial Ca→Cu substitution facilitated the release of constitutive hydroxyls and lowered the energy of formation of TCP from the apatite precursor at elevated temperatures. Addition of copper thus effectively inhibited the formation of apatite as the secondary phase. The copper-doped cement exhibited an antibacterial effect, though exclusively against gram-negative bacteria, including E. coli, P. aeruginosa and S. enteritidis. This antibacterial effect was due to copper ions, as demonstrated by an almost negligible antibacterial effect of the pure, copper-free cement. Also, the antibacterial activity of the copper-containing cement was significantly higher than that of its precursor powder. Since there was no significant difference between the kinetics of the release of copper from the precursor TCP powder and from the final, brushite phase of the hardened cement, this has suggested that the antibacterial effect was not solely due to copper ions, but due to the synergy between cationic copper and a particular phase and aggregation state of calcium phosphate. Though inhibitory to bacteria, the copper-doped cement increased the viability of human glial E297 cells, murine osteoblastic K7M2 cells and especially human primary lung fibroblasts. That this effect was also due to copper ions was evidenced by the null effect on viability increase exhibited by the copper-free cements. The difference in the mechanism of protection of dehydratases in prokaryotes and eukaryotes was used as a rationale for explaining the hereby evidenced selectivity in biological response. It presents the basis for the consideration of copper as a dually effective ion when synergized with calcium phosphates: toxic for bacteria and beneficial for the healthy cells.

Succeeding in the substitution of pharmaceutical compounds with ions deliverable with the use of resorbable biomaterials could have far-reaching benefits for medicine and economy. Calcium phosphates are known as excellent accommodators of foreign ions. Manganese, the fifth most abundant metal on Earth was studied here as an ionic dopant in β-tricalcium phosphate (β-TCP) ceramics. β-TCP containing different amounts of Mn2+ ions per Mn x Ca3–x (PO4)2 formula (x = 0, 0.001, 0.01, and 0.1) was investigated for a range of physicochemical and biological properties. The results suggested the role of Mn2+ as a structure booster, not breaker. Mn2+ ions increased the size of coherent X-ray scattering regions averaged across all crystallographic directions and also lowered the temperature of transformation of the hydroxyapatite precursor to β-TCP. The particle size increased fivefold, from 20 to 100 nm, in the 650–750 °C region, indicating that the reaction of formation of β-TCP was accompanied by a considerable degree of grain growth. The splitting of the antisymmetric stretching mode of the phosphate tetrahedron occurred proportionally to the Mn2+ content in the material, while electron paramagnetic resonance spectra suggested that Mn2+ might substitute for three out of five possible calcium ion positions in the unit cell of β-TCP. The biological effects of Mn-free β-TCP and Mn-doped β-TCP were selective: moderately proliferative to mammalian cells, moderately inhibitory to bacteria, and insignificant to fungi. Unlike pure β-TCP, β-TCP doped with the highest concentration of Mn2+ ions significantly inhibited the growth of all bacterial species tested: Staphylococcus aureus, Salmonella typhi, Escherichia coli, Pseudomonas aeruginosa, and Enterococcus faecalis. The overall effect against the Gram-positive bacteria was more intense than against the Gram-negative microorganisms. Meanwhile, β-TCP alone had an augmentative effect of the viability of adipose-derived mesenchymal stem cells (ADMSCs) and the addition of Mn2+ tended to reduce the extent of this augmentative effect, but without imparting any toxicity. For all Mn-doped β-TCP concentrations except the highest, the cell viability after 72 h incubation was significantly higher than that of the negative control. Assays evaluating the effect of Mn2+-containing β-TCP formulations on the differentiation of ADMSCs into three different lineagesosteogenic, adipogenic, and chondrogenicdemonstrated no inhibitory or adverse effects compared to pure β-TCP and powder-free positive controls. Still, β-TCP delivering the lowest amount of Mn2+ seemed most effective in sustaining the differentiation process toward all three phenotypes, indicating that the dose of Mn2+ in β-TCP need not be excessive to be effective.

The mineral constituent of bone tissue is a carbonate-substituted apatite (CHA). The thermal stability of the CHA has been revealed to depend on the substitution type and degree, although relatively little is known about this behavior. The aim of this study was to investigate the carbonate loss from synthetic CHAs in equilibrium conditions in a wide temperature range. An approach based on FTIR spectroscopy of condensed gas phase was applied to evaluate the CO and CO2 release with increasing temperature. Four different CHAs were studied, which were prepared by either precipitation from solution or the solid-state interaction. The samples differ from each other by the substitution degree. In one of the samples calcium was partially substituted by magnesium. Decomposition was shown to start at surprisingly low temperature, about 400 degrees C, and the CO content increases monotonously with the increase of temperature. The CO2 content goes through a maximum due to its decomposition into carbon monoxide and oxygen, the temperature of this maximum being strongly dependent on the chemical synthesis route. Therefore, control of the sintering atmosphere with respect to the CO2/CO ratio is needed when preparing the carbonated apatite bioceramics.

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