One-Pot Synthesis of Bioinspired Peptide-Decorated Apatite Nanoparticles for Nanomedicine.

Hybrid organic-inorganic bio-inspired apatite nanoparticles (NPs) are attractive for biomedical applications and especially in nanomedicine. Unfortunately, their applications in nanomedicine are limited by their broad particle size distributions and uncontrolled drug loading due to their multistep synthesis process.  Besides, very few attempts at exposing bioactive peptides on apatite NPs are made. In this work, an original one-pot synthesis of well-defined bioactive hybrid NPs composed of a mineral core of bioinspired apatite surrounded by an organic corona of bioactive peptides is reported. Dual stabilizing-bioactive agents, phosphonated polyethylene glycol-peptide conjugates, are prepared and directly used during apatite precipitation i) to form the organic corona during apatite precipitation, driving the size and shape of resulting hybrid NPs with colloidal stabilization and ii) to expose peptide moieties (RGD or YIGSR sequences) at the NPs periphery in view of conferring additional surface properties to enhance their interaction with cells. Here, the success of this approach is demonstrated, the functionalized NPs are fully characterized by Fourier-transform infrared, Raman, X-ray diffraction, solid and liquid state NMR, transmission electron microscopy, and dynamic light scattering, and their interaction with fibroblast cells is followed, unveiling a synergistic proliferative effect.

Impact of Polyethylenimine Conjugation Mode on the Cell Transfection Efficiency of Silica Nanovectors.

The conjugation of polyethylenimine (PEI) to silica nanoparticles has emerged as a useful strategy in gene delivery. Here we investigate the influence of the PEI conjugation mode on the transfection ability of plain silica nanoparticles. Surface functionalization with sulfonate- and chloride-bearing silanes modulates the amount and conformation of PEI and therefore the particles' affinity for the plasmid, without impacting on cytotoxicity. However, transfection efficiency in both immortalized and primary cells is more directly correlated to the nature and strength of the particle-PEI interactions. It suggests that PEI detachment from the particle surface at the stage of endosomal escape is a key event in the plasmid delivery process. These data should provide fruitful guidelines for the fine tuning of colloidal surfaces intended for intracellular delivery of bioactive molecules.

Water-mediated structuring of bone apatite.

It is well known that organic molecules from the vertebrate extracellular matrix of calcifying tissues are essential in structuring the apatite mineral. Here, we show that water also plays a structuring role. By using solid-state nuclear magnetic resonance, wide-angle X-ray scattering and cryogenic transmission electron microscopy to characterize the structure and organization of crystalline and biomimetic apatite nanoparticles as well as intact bone samples, we demonstrate that water orients apatite crystals through an amorphous calcium phosphate-like layer that coats the crystalline core of bone apatite. This disordered layer is reminiscent of those found around the crystalline core of calcified biominerals in various natural composite materials in vivo. This work provides an extended local model of bone biomineralization.

High dynamics and precision optical measurement using a position sensitive detector (PSD) in reflection-mode: application to 2D object tracking over a Smart Surface.

When related to a single and good contrast object or a laser spot, position sensing, or sensitive, detectors (PSDs) have a series of advantages over the classical camera sensors, including a good positioning accuracy for a fast response time and very simple signal conditioning circuits. To test the performance of this kind of sensor for microrobotics, we have made a comparative analysis between a precise but slow video camera and a custom-made fast PSD system applied to the tracking of a diffuse-reflectivity object transported by a pneumatic microconveyor called Smart-Surface. Until now, the fast system dynamics prevented the full control of the smart surface by visual servoing, unless using a very expensive high frame rate camera. We have built and tested a custom and low cost PSD-based embedded circuit, optically connected with a camera to a single objective by means of a beam splitter. A stroboscopic light source enhanced the resolution. The obtained results showed a good linearity and a fast (over 500 frames per second) response time which will enable future closed-loop control by using PSD.

Investigation of the interface in silica-encapsulated liposomes by combining solid state NMR and first principles calculations.

In the context of nanomedicine, liposils (liposomes and silica) have a strong potential for drug storage and release schemes: such materials combine the intrinsic properties of liposome (encapsulation) and silica (increased rigidity, protective coating, pH degradability). In this work, an original approach combining solid state NMR, molecular dynamics, first principles geometry optimization, and NMR parameters calculation allows the building of a precise representation of the organic/inorganic interface in liposils. {(1)H-(29)Si}(1)H and {(1)H-(31)P}(1)H Double Cross-Polarization (CP) MAS NMR experiments were implemented in order to explore the proton chemical environments around the silica and the phospholipids, respectively. Using VASP (Vienna Ab Initio Simulation Package), DFT calculations including molecular dynamics, and geometry optimization lead to the determination of energetically favorable configurations of a DPPC (dipalmitoylphosphatidylcholine) headgroup adsorbed onto a hydroxylated silica surface that corresponds to a realistic model of an amorphous silica slab. These data combined with first principles NMR parameters calculations by GIPAW (Gauge Included Projected Augmented Wave) show that the phosphate moieties are not directly interacting with silanols. The stabilization of the interface is achieved through the presence of water molecules located in-between the head groups of the phospholipids and the silica surface forming an interfacial H-bonded water layer. A detailed study of the (31)P chemical shift anisotropy (CSA) parameters allows us to interpret the local dynamics of DPPC in liposils. Finally, the VASP/solid state NMR/GIPAW combined approach can be extended to a large variety of organic-inorganic hybrid interfaces.

Ultrasound-assisted synthesis of mesoporous zirconia-hydroxyapatite nanocomposites and their dual surface affinity for Cr3+/Cr2O7(2-) ions.

Zirconia-hydroxyapatite nanocomposites were prepared by sol-gel deposition of zirconium oxide from a zirconium alkoxide in the presence of apatite colloidal suspension under ultrasonication. The material porosity evolves from mainly microporous zirconia to mesoporous hydroxyapatite, with decreasing surface area and increasing pore volume. XRD studies indicate that the apatite phase is well-preserved within the composite materials. The homogeneous dispersion of apatite colloids within the zirconia network was supported by TEM observations and nitrogen sorption measurements. (31)P solid-state NMR studies suggest that partial dissolution of apatite may have occurred during the preparation, leading to the adsorption of phosphate species on zirconia particles. This is confirmed by XRD studies of nanocomposites after thermal treatment that demonstrate the preferred formation of tetragonal over monoclinic ZrO(2) in the presence of hydroxyapatite. In order to investigate the surface properties of these novel materials, the adsorption of Pb(2+), Cr(3+), and Cr(2)O(7)(2-) was evaluated. Metal cations were preferentially adsorbed on apatite-rich composites, whereas Cr(2)O(7)(2-) shows a good affinity for the zirconia-rich phases. Zirconia-apatite materials showed the most promising performance in terms of recyclability. These nanocomposites that combine microporosity, mesoporosity and dual sorption properties for these species appear as interesting materials for metal ion remediation and may also find applications as biomaterials.

Baicalein-modified hydroxyapatite nanoparticles and coatings with antibacterial and antioxidant properties.

Aseptic loosening and bacterial infections are the two main causes of failure for metallic implants used for joint replacement. A coating that is both bioactive and possesses antimicrobial properties may address such shortcomings and improve the performance of the implant. We have sought to study the properties of combining hydroxyapatite-based nanoparticles or coatings with baicalein, a plant-extracted molecule with both antibacterial and antioxidant properties. (B-type) carbonated hydroxyapatite nanoparticles prepared by a chemical wet method could subsequently adsorbed by soaking in a baicalein solution. The amount of adsorbed baicalein was determined to be 63 mg.g-1 by thermogravimetric measurements. In a second approach, baicalein was adsorbed on a biomimetic calcium-deficient hydroxyapatite planar coating (12 µm thick) deposited on Ti6Al4V alloy from an aqueous solution of calcium, phosphate, sodium and magnesium salts. Soaking of the hydroxyapatite coated on titanium alloy in a baicalein solution induced partial dissolution/remodeling of the upper surface of the coating. However, the observed remodeling of the surface was much more pronounced in the presence of a baicalein solution, compared to pure water. The presence of adsorbed baicalein on the HAp layer, although it could not be precisely quantified, was assessed by XPS and fluorescence analysis. Planar coatings exhibited significant antibacterial properties against Staphylococcus epidermidis. Baicalein-modified nanoparticles exhibited significant antioxidant properties. These results illustrate the potential of hydroxyapatite used as a carrier for natural biologically-active molecules and also discuss the challenges associated with their applications as antibacterial agents.

A soft-chemistry approach to the synthesis of amorphous calcium ortho/pyrophosphate biomaterials of tunable composition.

The development of amorphous phosphate-based materials is of major interest in the field of biomaterials science, and especially for bone substitution applications. In this context, we herein report the synthesis of gel-derived hydrated amorphous calcium/sodium ortho/pyrophosphate materials at ambient temperature and in water. For the first time, such materials have been obtained in a large range of tunable orthophosphate/pyrophosphate molar ratios. Multi-scale characterization was carried out thanks to various techniques, including advanced multinuclear solid state NMR. It allowed the quantification of each ionic/molecular species leading to a general formula for these materials: [(Ca2+y Na+z H+3+x-2y-z)(PO43-)1-x(P2O74-)x](H2O)u. Beyond this formula, the analyses suggest that these amorphous solids are formed by the aggregation of colloids and that surface water and sodium could play a role in the cohesion of the whole material. Although the full comprehension of mechanisms of formation and structure is still to be investigated in detail, the straightforward synthesis of these new amorphous materials opens up many perspectives in the field of materials for bone substitution and regeneration. STATEMENT OF SIGNIFICANCE: The metastability of amorphous phosphate-based materials with various chain length often improves their (bio)chemical reactivity. However, the control of the ratio of the different phosphate entities has not been yet described especially for small ions (pyrophosphate/orthophosphate) and using soft chemistry, whereas it opens the way for the tuning of enzyme- and/or pH-driven degradation and biological properties. Our study focuses on elaboration of amorphous gel-derived hydrated calcium/sodium ortho/pyrophosphate solids at 70 °C with a large range of orthophosphate/pyrophosphate ratios. Multi-scale characterization was carried out using various techniques such as advanced multinuclear SSNMR (31P, 23Na, 1H, 43Ca). Analyses suggest that these solids are formed by colloids aggregation and that the location of mobile water and sodium could play a role in the material cohesion.

Role of carboxylate chelating agents on the chemical, structural and textural properties of hydroxyapatite.

Organically-modified hydroxyapatite materials were synthesized through the addition of oxalic, succinic, adipic and citric acids to a calcium hydroxide solution before neutralization by ammonium dihydrogenphosphate. All carboxylic acids have a significant influence on apatite crystallinity and nanoparticle size, as indicated by XRD and TEM. Chemical and thermogravimetric analyses as well as FTIR and {(1)H}-(13)C CP MAS NMR spectroscopies indicate that the additives are present in the final material. (1)H, {(1)H}-(31)P HPDec MAS, CP MAS and 2D {(1)H}-(31)P CP-HETCOR MAS NMR experiments suggest that carboxylic acids are localized on the apatite nanocrystallite surface, resulting in the formation of a disordered outer layer. Nitrogen sorption measurements indicate minor modifications of the specific surface area of the resulting mesoporous materials upon carboxylic acid addition but more significant variations in the average dimensions of the pores as well as in the chemical nature of the pore surface. Although these evolutions are mainly in good agreement with the ligand affinity for calcium ions in solution, an unexpected difference was observed between succinic and adipic acid, that may be attributed to steric constraints resulting from the interfacial nature of the calcium-ligand interactions. These data should provide useful guidelines to identify novel efficient additives to control apatite growth.