CIRIMAT

The incorporation of foreign ions, such as Mg 2+ , exhibiting a biological activity for bone regeneration is presently considered as a promising route for increasing the bioactivity of bone-engineering scaffolds. In this work, the morphology, structure, and surface hydration of biomimetic nanocrystalline apatites were investigated before and after surface exchange with such Mg 2+ ions, by combining chemical alterations (ion exchange, H 2 O-D 2 O exchanges) and physical examinations (Fourier transform infrared spectroscopy (FTIR) and highresolution transmission electron microscopy (HRTEM)). HRTEM data suggested that the Mg 2+ /Ca 2+ exchange process did not affect the morphology and surface topology of the apatite nanocrystals significantly, while a new phase, likely a hydrated calcium and/or magnesium phosphate, was formed in small amount for high Mg concentrations. Near-infrared (NIR) and medium-infrared (MIR) spectroscopies indicated that the samples enriched with Mg 2+ were found to retain more water at their surface than the Mg-free sample, both at the level of H 2 O coordinated to cations and adsorbed in the form of multilayers. Additionally, the H-bonding network in defective subsurface layers was also noticeably modified, indicating that the Mg 2+ /Ca 2+ exchange involved was not limited to the surface. This work is intended to widen the present knowledge on Mg-enriched calcium phosphate-based bioactive materials intended for bone repair applications.

In order to shed some light on DNA preservation over time in skeletal remains from a physicochemical viewpoint, adsorption and desorption of DNA on a well characterized synthetic apatite mimicking bone and dentin biominerals were studied. Batch adsorption experiments have been carried out to determine the effect of contact time (kinetics), DNA concentration (isotherms) and environmentally relevant factors such as temperature, ionic strength and pH on the adsorption behavior. The analogy of the nanocrystalline carbonated apatite used in this work with biological apatite was first demonstrated by XRD, FTIR, and chemical analyses. Then, DNA adsorption kinetics was fitted with the pseudo-first order, pseudo-second order, Elovich, Ritchie and double exponential models. The best results were achieved with the Elovich kinetic model. The adsorption isotherms of partially sheared calf thymus DNA conformed satisfactorily to Temkin's equation which is often used to describe heterogeneous adsorption behavior involving polyelectrolytes. For the first time, the irreversibility of DNA adsorption toward dilution and significant phosphate-promoted DNA desorption were evidenced, suggesting that a concomitant ion exchange process between phosphate anionic groups of DNA backbone and labile non-apatitic hydrogenphosphate ions potentially released from the hydrated layer of apatite crystals. This work should prove helpful for a better understanding of diagenetic processes related to DNA preservation in calcified tissues.

Carbonated apatites represent an important class of compounds encountered in many fields including anthropology, archeology, geology, medicine and biomaterials engineering. They constitute, in particular, the mineral part of bones and teeth, are found in sedimentary settings, and are used as biomimetic compounds for the development of bone tissue engineering scaffolds. Whether for assessing the degree of biomimetism of synthetic apatites or for better understanding diagenetic events, their thorough physico-chemical characterization is essential, and includes, in particular, the evaluation of their carbonate content. FTIR is especially well-suited for such a goal, as this spectroscopy technique requires only a low amount of specimen to analyze, and carbonate ions exhibit a clear vibrational signature. In this contribution, we critically discuss several FTIR-approaches that may be (or have been) considered in view of carbonation quantification. The best methodology appears to be based on the analysis of the n 3 (CO 3 ) and n 1 n 3 (PO 4 ) modes. The area ratio r c/p between these two contributions was found to be directly correlated to the carbonate content of the samples (R 2 ¼ 0.985), with the relation wt.% CO 3 ¼ 28.62*r c/ p þ 0.0843. The method was validated thanks to titrations by coulometry assays for various synthetic reference samples exhibiting carbonate contents between 3 and 7 wt.%. The FTIR carbonate quantification methodology that we propose here was also tested with success on three skeletal specimens (two bones/one tooth), after elimination of the collagen contribution. Comparative data analysis is also presented, showing that the use of other vibration bands, or only peak heights (instead of peak areas), leads to significantly lower correlation agreement. This FTIR data treatment methodology is recommended so as to limit errors on the evaluation of carbonate contents in apatite substrates.

Nanocrystalline apatites analogous to bone mineral are very promising materials for the preparation of highly bioactive ceramics due to their unique intrinsic physico-chemical characteristics. Their surface reactivity is indeed linked to the presence of a metastable hydrated layer on the surface of the nanocrystals. Yet the sintering of such apatites by conventional techniques, at high temperature, strongly alters their physico-chemical characteristics and biological properties, which points out the need for ''softer" sintering processes limiting such alterations. In the present work a non-conventional technique, spark plasma sintering, was used to consolidate such nanocrystalline apatites at non-conventional, very low temperatures (T < 300°C) so as to preserve the surface hydrated layer present on the nanocrystals. The bioceramics obtained were then thoroughly characterized by way of complementary techniques. In particular, microstructural, nanostructural and other major physico-chemical features were investigated and commented on. This work adds to the current international concern aiming at improving the capacities of present bioceramics, in view of elaborating a new generation of resorbable and highly bioactive ceramics for bone tissue engineering.

Intracellular drug delivery using colloidal biomimetic calcium phosphate apatites as nanocarriers is a seducing concept. However, the colloid preparation to an industrial scale requires the use of easily handled raw materials as well as the possibility to tailor the nanoparticles size. In this work, the stabilization of the colloids was investigated with various biocompatible agents. Most interestingly, nanoscale colloids were obtained without the need for toxic and/or hazardous raw materials. Physico-chemical characteristics were investigated by chemical analyses, dynamic light scattering, FTIR/Raman spectroscopies, XRD, and electron microscopy. A particularly promising colloidal system associates biomimetic apatite stabilized with a natural phospholipid moiety (AEP r , 2-aminoethylphosphoric acid).

Luminescent colloidal nanosystems based on europium-doped biomimetic apatite were prepared and investigated. The colloids were synthesized by soft chemistry in the presence of a phospholipid moiety, 2-aminoethylphosphoric acid (AEP), with varying europium doping rates. Physicochemical features, including compositional, structural, morphological, and luminescence properties, were examined. Experimental evidence showed that suspensions prepared from an initial Eu/(Eu + Ca) molar ratio up to 2% consisted of singlephased biomimetic apatite nanocrystals covered with AEP molecules. The mean particle size was found to depend closely on the AEP content, enabling the production of apatite colloids with a controlled size down to ca. 30 nm. The colloids showed luminescence properties typical of europium-doped systems with narrow emission bands and long luminescence lifetimes of the order to the millisecond, and the data suggested the location of Eu 3+ ions in a common crystallographic environment for all the colloids. These systems, stable over time and capable of being excited in close-to-visible or visible light domains, may raise interest in the future in the field of medical imaging. * To whom correspondence should be addressed. Phone: +33 (0)5 34 32 34 50 (A.A.), +33 (0)5 34 32 34 11 (C.D.). Fax: +33 (0)5 34 32 33 99 (A.A.), +33 (0)5 34 32 33 99 (C.D.).

The modification of the composition of apatite materials can be made by several processes corresponding to ion exchange reactions which can conveniently be adapted to current coatings and ceramics and are an alternative to setting up of new synthesis methods. In addition to high temperature thermal treatments, which can partly or almost totally replace the monovalent OH − anion of stoichiometric hydroxyapatite by any halogen ion or carbonate, aqueous processes corresponding to dissolution-reprecipitation reactions have also been proposed and used. However, the most interesting possibilities are provided by aqueous ion exchange reactions involving nanocrystalline apatites. These apatites are characterised by the existence on the crystal surface of a hydrated layer of loosely bound mineral ions which can be easily exchanged in solution. This layer offers a possibility to trap mineral ions and possibly active molecules which can modify the apatite properties. Such processes are involved in mineralised tissues and could be used in biomaterials for the release of active mineral species. C 2005 Springer Science + Business Media, Inc.

Nanocrystalline apatitic calcium phosphates play a crucial role in calcified tissues and biomaterials. One of the most interesting characteristics of biomimetic apatite nanocrystals is the existence of a surface hydrated layer essentially related to their formation process in solution. This hydrated layer shows specific spectroscopic characteristics. It seems to exist in its nascent state only in wet samples and is altered on drying. This surface-hydrated layer progressively disappears as the stable apatite domains develop. The surface ions can be rapidly and reversibly exchanged in solution, mainly with selected bivalent species. The exchange reactions clearly reveal the existence of two domains: the relatively inert apatite core and the very reactive surface-hydrated domains. The structure of the hydrated layer has been shown to be reversibly affected by the constituting ions. Such a surface layer in bone apatite nanocrystals could participate actively in homeostasis and probably other regulation processes. The specificity of biomimetic apatite nano-crystals also opens interesting possibilities in materials science. The mobility of the mineral ions on the crystal surface, for example, allows strong bonding and interactions either with other crystals or with different substrates. Inter-crystalline interactions have been described as a "crystal fusion" process in vivo and they could be involved in the setting reaction of biomimetic calcium phosphate cements. Ceramic-like materials using the surface interaction capabilities of the nanocrystals can be produced at very low temperature (below 200 C). The surface-hydrated layer could also be involved in interactions with macromolecules and polymeric materials or in the coating of implants. The ion exchange and adsorption capabilities of the nanocrystals could probably be used for drug release, offering a range of possible behaviours.

Biological mineralisations based on calcium phosphate. The mineral fractions of bones and teeth are non-stoichiometric apatite crystals. Their morphology, dimensions, composition and reactivity are adapted to their biological function. Bone apatites are made of very reactive nanocrystals possessing, on their surface, a structured hydrated layer responsible for their biological properties. This unstable layer becomes progressively and unavoidably transformed into a

Nonstoichiometric nickel-copper spinel manganites were found to be highly active for the reduction of nitric oxide by carbon monoxide at low temperature (300 • C). The activity increases with the level of nonstoichiometry of the oxides and almost linearly with their specific surface area. Moreover, this activity was found to depend strongly on the copper content, nickel manganites being almost inactive. The influence of the surface state of the oxides was also investigated. Cuprous cations are thought to play the determining role in the catalytic cycle.