Biomimetic apatite-based biomaterials: on the critical impact of synthesis and post-synthesis parameters

Nanocrystalline apatites are major constituents of hard tissues, and attempts are made worldwide to prepare synthetic analogs. However the impact of synthesis/ postsynthesis parameters is often disregarded. Based on an updated knowledge on such compounds, we inspected the effects of synthesis parameters (maturation time, temperature,pH, nature of counter-ions) and post-treatments (re-immersion in aqueous media, thermal treatment) on physicochemical characteristics. Great modifications were noticed during the 3 first days of maturation, where a progressive evolution of the apatite phase (localized in the core of the nanocrystals) toward stoichiometry was observed at the expense of the non-apatitic surface layer which progressively disappears. Similar trends were also evidenced for maturation run under increasing temperatures, studied here in the range 20–100 °C. pH impacted more specifically the chemical composition. The nature of the counter-ion in the starting phosphate salt influenced composition and nonstoichiometry, depending on its (in)ability to be incorporated in the lattice. Freeze-drying allowed to preserve a high surface reactivity, although further evolutions were noticed after re-immersion. Effects of a thermal treatment of dried samples were unveiled, suggesting a denaturation of the hydrated layer on the nanocrystals. This work underlines the necessity of a good control of synthesis/postsynthesis parameters for the production of biomimetic apatites

Peroxide-doped apatites: Preparation and effect of synthesis parameters

This contribution focused on the preparation of peroxide-doped calciumphosphate apatites—in viewof potential uses as bioactive bioceramics with antimicrobial functions, and on their main physico-chemical characteristics. Two synthesis routeswere investigated. First, the hydrolysis of β-TCP in the presence ofH2O2 was followed. However, only elevated concentrations inH2O2 in themediumor temperatures around 150 °C allowed us to reach the complete β-TCP-to-apatite hydrolysis process, and the obtained samples exhibited a high crystallinity state with no non-apatitic chemical environments. The second protocol tested consisted in the direct apatite precipitation in the presence of H2O2 in themedium (at room temperature). This protocol led to single-phased nanocrystalline apatites, and our data indicate that part of the apatitic OH− ions were substituted by oxygenated species, and typically by peroxide ions (quantified). Physico-chemical modifications in the form of an improvement in crystallinity state, an increase in unit cell volume, and the presence of additional Raman bands were noticed and discussed

Enzyme-functionalized biomimetic apatites: concept and perspectives in view of innovative medical approaches

Biomimetic nanocrystalline calcium-deficient apatite compounds are particularly attractive for the setup of bioactive bone-repair scaffolds due to their high similarity to bone mineral in terms of chemical composition, structural and substructural features. As such, along with the increasingly appealing development of moderate temperature engineered routes for sample processing, they have widened the armamentarium of orthopedic and maxillofacial surgeons in the field of bone tissue engineering. This was made possible by exploiting the exceptional surface reactivity of biomimetic apatite nanocrystals, capable of easily exchanging ions or adsorbing (bio)molecules, thus leading to highly-versatile drug delivery systems. In this contribution we focus on the preparation of hybrid materials combining biomimetic nanocrystalline apatites and enzymes (lysozyme and subtilisin). This paper reports physico-chemical data as well as cytotoxicity evaluations towards Cal-72 osteoblast-like cells and finally antimicrobial assessments towards selected strains of interest in bone surgery. Biomimetic apatite/enzyme hybrids could be prepared in varying buffers. They were found to be non-cytotoxic toward osteoblastic cells and the enzymes retained their biological activity (e.g. bond cleavage or antibacterial properties) despite the immobilization and drying processes. Release properties were also examined. Beyond these illustrative examples, the concept of biomimetic apatites functionalized with enzymes is thus shown to be useable in practice, e.g. for antimicrobial purposes, thus widening possible therapeutic perspectives

Nanocrystalline apatites: The fundamental role of water

Bone is a natural nanocomposite. Its mineral component is nanocrystalline calcium phosphate apatite, whose synthetic biomimetic analogs can be prepared by wet chemistry. The initially formed crystals, whether biological or synthetic, exhibit very peculiar physicochemical features. In particular, they are nanocrystalline, nonstoichiometric, and hydrated. The surface of the nanocrystals is covered by a non-apatitic hydrated layer containing mobile ions, which may explain their exceptional surface reactivity. For their precipitation in vivo or in vitro, for their evolution in solution, for the 3D organization of the nanocrystals, and for their consolidation to obtain bulk ceramic materials, water appears to be a central component that has not received much attention. In this mini-review, we explore these key roles of water on the basis of physicochemical and thermodynamic data obtained by complementary tools including FTIR, XRD, ion titrations, oxide melt solution calorimetry, and cryo-FEG-SEM. We also report new data obtained by DSC, aiming to explore the types of water molecules associated with the nanocrystals. These data support the existence of two main types of water molecules associated with the nanocrystals, with different characteristics and probably different roles and functions. These findings improve our understanding of the behavior of bioinspired apatite-based systems for biomedi- cine and also of biomineralization processes taking place in vivo, at present and in the geologic past. This paper is thus intended to give an overview of the specificities of apatite nanocrystals and their close relationship with water

Elaboration and characterization of innovative osseous biomaterials based on doped calcium phosphate apatites

Les infections nosocomiales post-opératoires en sites osseux posent un problème majeur de santé publique. L'utilisation de biocéramiques bioactives et résorbables qui présenteraient des propriétés antibactériennes apparaît comme une des solutions les plus prometteuses pour lutter contre l'invasion de micro-organismes au niveau du site opératoire. Les apatites nanocristallines biomimétiques se révèlent être des candidats de choix pour ces applications en raison de leur similitude avec le minéral osseux et de leur forte réactivité de surface. Cependant, elles ne possèdent pas de propriétés antibactériennes intrinsèques, ce qui peut potentiellement être amené par un dopage ionique approprié. Dans ce contexte, ce travail traite de la synthèse et de la caractérisation physico-chimique d'apatites biomimétiques enrichies avec des cations zinc (Zn2+), cuivre (Cu2+) ou argent (Ag+) ou avec des anions oxygénés de type peroxydes qui présenteraient ces facultés antimicrobiennes ; puis sur l'évaluation préliminaire de leurs propriétés (micro)biologiques. Dans un premier temps, l'étude de systèmes apatitiques non dopés a indiqué qu'à l'instar du minéral osseux les apatites nanocristallines présentaient des cristaux de dimensions nanométriques dont la composition chimique s'éloignait de la stoechiométrie et exposant des environnements ioniques non-apatitiques hydratés en surface des nanocristaux. L'influence des paramètres de synthèse a été évaluée et révèle que le temps de maturation, la température, le pH et la nature des sels de phosphate impactent significativement les caractéristiques physico-chimiques de ces composés. Nous montrons également que les conditions de post-traitement (ré-immersion, traitement thermique, mise en forme) peuvent aussi modifier significativement les caractéristiques finales des biocéramiques. Dans un second temps, ce travail a révélé que l'enrichissement d'apatites nanocristallines avec des ions Zn2+, Cu2+, Ag+ ou des espèces oxygénées était possible – avec des taux de dopage limites qui ont été évalués – mais générait des modifications physico-chimiques notables en particulier en termes d'état de cristallinité et de teneur en environnements chimiques non-apatitiques. Le zinc et le cuivre engendrent des effets similaires et semblent agir en tant qu'inhibiteur de croissance cristalline. L'argent, bien que monovalent, ne modifie pas significativement les processus de formation et de croissance des nanocristaux d'apatites. En revanche, la présence de peroxyde d'hydrogène dans le milieu réactionnel conduit à la formation d'apatite dont l'état de cristallinité est augmenté. Le choix de paramètres de synthèse adéquats, influençant notablement les mécanismes d'incorporation des ions, s'est avéré déterminant pour l'obtention d'apatites nanocristallines monophasées et dopées. Des tests biologiques préliminaires ont été réalisés pour évaluer la cytotoxicité de ces composés et le comportement de cellules ostéogéniques (de types ostéoblastes et ostéoclastes). L'évaluation d'éventuelles propriétés antibactériennes a également fait l'objet de ce travail, dans le cadre d'une collaboration internationale. Parmi les formulations présentant des propriétés antibactériennes mesurées, les apatites biomimétiques enrichies en argent apparaissent au vu de ce travail comme les candidats les plus prometteurs pour conférer l'effet antibactérien nécessaire aux applications visées.Hospital acquired infections in osseous sites are a major issue of public health. The use of bioactive and resorbable bioceramics that present antibacterial properties appears as one of the most promising solution against microorganisms invasion of the surgical site. Biomimetic nanocrystalline apatites belong to the choice candidates for those applications thanks to their similarities with the bone mineral and their high surface reactivity. Nevertheless, they don't possess intrinsic antibacterial capacity, property that can be reached by a suitable ionic enrichment. In this context, this work deals with the synthesis and the physico-chemical characterization of such biomimetic nanocrystalline apatites doped with zinc (Zn2+), copper (Cu2+) or silver (Ag+) cations or with oxygenated anions like peroxides that would present antibacterial activity; then their (micro-)biological properties have also been studied. As a first part, study of apatitic non-doped systems shows that, as well as bone mineral, nanocrystalline apatites exhibit nano-sized crystals which chemical composition depart from stoichiometry and possess hydrated non-apatitic environments on their surface. The influence of synthesis parameters has been studied and reveals that maturation time in solution, temperature, pH or the nature of starting phosphate salts impact significantly the physico-chemical characteristics of those compounds. We also point out that post-treatment conditions (re-immersion, thermal treatment, forming) can significantly modify final characteristics of those bioceramics. As a second part, this work reveals that nanocrystalline apatites enrichment with Zn2+, Cu2+, Ag+ or oxygenated species seem to be possible – with maximal doping rates that were evaluated – but generate significant physico-chemical modifications, especially in terms of crystallinity state or non-apatitic chemical environments content. Zinc and copper act on a similar way on apatite compound and exhibit a crystal growth inhibitory role. Silver, even with a single positive charge, don't modify significantly the formation and the growth mechanisms of apatites nanocrystals. On the opposite, hydrogen peroxide presence in the synthesis media generates the formation of apatites which crystallinity state is improved. All of those results suggest that synthesis parameters are determining to obtain doped nanocrystalline apatites and that they influence notably the incorporation mechanisms of ions. Finally, preliminary biological tests have been realized in order to evaluate the cytotoxicity and the behavior of osteogenic cells (osteoblast and osteoclast type) in contact with those compounds. The evaluation of potential antibacterial properties is also discussed in this work as part of an international collaboration. Among formulations that exhibit measured antibacterial activity, silver doped biomimetic apatites appear as the most promising candidates to confer antibacterialness necessary for the envisaged applications

Synthesis and post-treatments of biomimetic apatites: How working conditions may configure final physico-chemical features

National audienceNanocrystalline apatites constitute the mineral part of hard tissues, and can be reproduced synthetically. Nonetheless,the impact of synthesis/post-synthesis parameters is often disregarded. Based on actualized knowledge on their physico-chemical features, we investigated these aspects on a systematic experimental basis. The apatite maturation state has a direct effect on the surface and core of the nanocrystals. Drying and re-immersion aspects were also examined in view of applications as implantable biomaterials: an equilibration of the samples surface is proposed to avoid acidification phenomena after re-immersion of drie

Synthesis and post-treatments of biomimetic apatites: How working conditions may configure final physico-chemical features

Nanocrystalline apatites constitute the mineral part of hard tissues, and can be reproduced synthetically. Nonetheless, the impact of synthesis/post-synthesis parameters is often disregarded. Based on actualized knowledge on their physico-chemical features, we investigated these aspects on a systematic experimental basis. The apatite maturation state has a direct effect on the surface and core of the nanocrystals. Drying and re-immersion aspects were also examined in view of applications as implantable biomaterials: an equilibration of the samples surface is proposed to avoid acidification phenomena after re-immersion of dried samples