A novel synthetic approach of cerium oxide nanoparticles with improved biomedical activity

Cerium oxide nanoparticles (CNPs) are novel synthetic antioxidant agents proposed for treating oxidative stress-related diseases. The synthesis of high-quality CNPs for biomedical applications remains a challenging task. A major concern for a safe use of CNPs as pharmacological agents is their tendency to agglomerate. Herein we present a simple direct precipitation approach, exploiting ethylene glycol as synthesis co-factor, to synthesize at room temperature nanocrystalline sub-10 nm CNPs, followed by a surface silanization approach to improve nanoparticle dispersibility in biological fluids. CNPs were characterized using transmission electron microscopy (TEM) observations, X-ray diffraction (XRD) analysis, thermogravimetric analysis (TGA), Fourier-transform infrared (FT-IR) spectroscopy, proton nuclear magnetic resonance (H-1-NMR) spectroscopy, dynamic light scattering (DLS) and zeta potential measurements. CNP redox activity was studied in abiotic systems using electron spin resonance (ESR) measurements, and in vitro on human cell models. In-situ silanization improved CNP colloidal stability, in comparison with non-functionalized particles, and allowed at the same time improving their original biological activity, yielding thus functionalized CNPs suitable for biomedical applications.

Synthesis and Characterization of Amphiphilic Gold Nanoparticles

Urszula Beata Cendrowska, Matteo Gasbarri, Zekiye Pelin Güven, Paulo Henrique Jacob Silva, Samuel Thomas Jones, Zhi Luo, Francesco Stellacci

Gold nanoparticles covered with a mixture of 1-octanethiol (OT) and 11-mercapto-1-undecane sulfonic acid (MUS) have been extensively studied because of their interactions with cell membranes, lipid bilayers, and viruses. The hydrophilic ligands make these particles colloidally stable in aqueous solutions and the combination with hydrophobic ligands creates an amphiphilic particle that can be loaded with hydrophobic drugs, fuse with the lipid membranes, and resist nonspecific protein adsorption. Many of these properties depend on nanoparticle size and the composition of the ligand shell. It is, therefore, crucial to have a reproducible synthetic method and reliable characterization techniques that allow the determination of nanoparticle properties and the ligand shell composition. Here, a one-phase chemical reduction, followed by a thorough purification to synthesize these nanoparticles with diameters below 5 nm, is presented. The ratio between the two ligands on the surface of the nanoparticle can be tuned through their stoichiometric ratio used during synthesis. We demonstrate how various routine techniques, such as transmission electron microscopy (TEM), nuclear magnetic resonance (NMR), thermogravimetric analysis (TGA), and ultraviolet-visible (UV-Vis) spectrometry, are combined to comprehensively characterize the physicochemical parameters of the nanoparticles.

JOURNAL OF VISUALIZED EXPERIMENTS2019

Etude par microscopie électronique de particules nanostructurées d'oxalate de cobalt précipitées en milieu aqueux

The understanding of material self-assembling and self-organization mechanisms, at mesoscale, is crucial for nanotechnologies development. Such structures, hierarchically organized in superlattices or colloïdal crystals, are observed in inorganic or organo-metallic precipitates. The classical characterization, using electron microscopy and X-ray diffraction, of a synthesized powder is inadequate to describe the hierarchically organized polycrystalline structure of the final particles. Here, we have highlighted the successive steps that lead to the spontaneous formation of a cobalt oxalate dihydrate (COD) colloïdal crystals that precipitate in aqueous media. The characterization of the final particles, made by X-ray and electronic diffraction, has allowed us to determine the controversial crystalline phase of the precipitated COD. The β and γ COD phases has been discriminated by comparing experimental results with simulations of X-ray diffractogram and electron diffraction patterns for these both structures. The precipitated COD corresponds to the γ phase indicating that it comes from solid dehydration of cobalt oxalate tetrahydrate. The final powder characterization, performed by X-ray diffraction (XRD), atomic force microscopy (AFM), low voltage high resolution scanning electron microscopy (LVHRSEM) and transmission electron microscopy (TEM), gives some diverging results about monocrystalline or polycrystalline nature of the precipitates. XRD and AFM analysis show a polycrystalline structure of particles whereas LVHRSEM, TEM and electron diffraction observations indicate a monocrystalline structure. This apparent contradiction has led us to elaborate a new characterization method allowing to follow the growth and the structural evolution of the precipitate as a function of time. The cryo- preparation techniques are well adapted as they allow freezing and direct observation of solid state suspension by electron microscopy. The freeze/drying method has been modified and used for both SEM and TEM, and has allowed us to study the morphological and structural evolution of the COD precipitate from nanoscale to mesoscale. In addition, TEM analysis of samples prepared by classical techniques has been performed to complement cryo-electron microscopy observations. The combination of these two methods show that COD precipitation is a complex multi-step process leading to the formation of a core/shell heterostructure. The anisotropic core is porous and partially crystalline. It is formed by agglomeration of isolated primary particles and agregated ones (15-20 nm sized secondary particles). The crystalline shell corresponds to the final particles faces and is made up by the layer by layer self-assembling and alignment of 5-7 nm sized crystalline primary particles. These nanoparticles are aligned and perfectly ordered in strings of nanograins in the layer structure. The self-assembling occurs in the last step of growth where we have a lower ionic strength and supersaturation inducing slower kinetics of growth and aggregation. Moreover, the crystalline order in the primary and secondary particles increase with the time of reaction consistent with a continuous process of primary particle nucleation and growth. Our results show that self-assembling occurs layer by layer with terraces, kinks and steps that are present on the faces. This reminds the layer by layer crystalline growth models but in this case, the buiding blocks are colloïdal instead of atomic or molecular. Based on these different results, we propose an original model describing the COD precipitate growth. In addition, we have studied the poly(methacrylic acid-sodium salt) effect on the COD precipitation. This additive (PMMA) appeared to be a very good growth inhibitor. The PMMA effect on the final particles morphology is dramatic. Its concentration variation in solution slows down the nanoparticles aggregation rates along a [101] preferential direction. The crystal habit can then be controlled and we have synthesized platelets, cubes and rods with tailored length. The use of cryogenic electron microscopy has been shown to be a powerful tool for the understanding of time dependent aspects of particle growth. The use of such techniques for the study of other systems, where the final precipitate appearance may hide particle substructures, will help to elucidate growth mechanisms and allow finer control of nanostructured materials for many applications. The complex self-assembling and self-organization processes could then be highlighted and studied on a nanometer to micrometer scale.

EPFL2004

Synthesis of Pyridyl Disulfide-Functionalized Nanoparticles for Conjugating Thiol-Containing Small Molecules, Peptides, and Proteins

Urara Hasegawa, Jeffrey Alan Hubbell

Previously we reported emulsion polymerization of propylene sulfide with Pluronic F127 as an emulsifier, yielding nanoparticles (NPs) in the 25 nm size range. Immunologically functional NPs were prepared by adding an antigen Pluronic conjugate to the polymerization mixture (Reddy, S. T., et al. (2007) Nat. Biotechnol. 25, 1159). We sought a more flexible scheme for conjugation of antigens and other biomolecules to the NP surfaces that would allow for milder reaction conditions than achievable during the polymerization step. Here, we present the synthesis of such functionalizable NPs in the form of NPs that carry thiol-reactive groups, to which thiol-containing antigens (peptide or protein) or other biomolecules can be conjugated under mild conditions to yield immunofunctional NPs. The Pluronic-stabilized poly(propylene sulfide) (PPS) NPs with thiol-reactive pyridyl disulfide groups are prepared in two steps by (1) emulsion polymerization of propylene sulfide in the presence of a carboxylate-Pluronic and (2) reaction of the carboxylic acid groups on the NP surface with cysteaminc pyridyl disulfide and a water-soluble carbodiimide reagent. We choose pyridyl disulfide groups to have a reduction-sensitive disulfide bond linking the antigen to the NP surface, allowing efficient release of antigen inside the cell in response to the reductive conditions within the endosome. The functionalizable NPs are characterized by proton NMR, dynamic light scattering (DLS), UV/vis spectroscopy, and transmission electron microscopy (TEM). Conjugation of small molecules and protein to the NP surface is presented.

2010

Etude par microscopie électronique de particules nanostructurées d'oxalate de cobalt précipitées en milieu aqueux

EPFL2004