| Centre
| | | | | | | webmail : intra-extra| Accès VPN| Accès IST
Univ. Paris-Saclay

Nina Královič-Kanjaková, Ali Asi Shirazi, Lukáš Hubčík, Mária Klacsová, Atoosa Keshavarzi, Juan Carlos Martínez, Sophie Combet, José Teixeira, and Daniela Uhríková

The use of an exogenous pulmonary surfactant (EPS) to deliver other relevant drugs to the lungs is a promising strategy for combined therapy. We evaluated the interaction of polymyxin B (PxB) with a clinically used EPS, the poractant alfa Curosurf (PSUR). The effect of PxB on the protein-free model system (MS) composed of four phospholipids (diC16:0PC/16:0–18:1PC/16:0–18:2PC/16:0–18:1PG) was examined in parallel to distinguish the specificity of the composition of PSUR. We used several experimental techniques (differential scanning calorimetry, small- and wide-angle X-ray scattering, small-angle neutron scattering, fluorescence spectroscopy, and electrophoretic light scattering) to characterize the binding of PxB to both EPS. Electrostatic interactions PxB–EPS are dominant. The results obtained support the concept of cationic PxB molecules lying on the surface of the PSUR bilayer, strengthening the multilamellar structure of PSUR as derived from SAXS and SANS. A protein-free MS mimics a natural EPS well but was found to be less resistant to penetration of PxB into the lipid bilayer. PxB does not affect the gel-to-fluid phase transition temperature, Tm, of PSUR, while Tm increased by ∼+ 2 °C in MS. The decrease of the thickness of the lipid bilayer (dL) of PSUR upon PxB binding is negligible. The hydrophobic tail of the PxB molecule does not penetrate the bilayer as derived from SANS data analysis and changes in lateral pressure monitored by excimer fluorescence at two depths of the hydrophobic region of the bilayer. Changes in dL of protein-free MS show a biphasic dependence on the adsorbed amount of PxB with a minimum close to the point of electroneutrality of the mixture. Our results do not discourage the concept of a combined treatment with PxB-enriched Curosurf. However, the amount of PxB must be carefully assessed (less than 5 wt % relative to the mass of the surfactant) to avoid inversion of the surface charge of the membrane


On De Gennes narrowing of fluids confined at the molecular scale in nanoporous materials
Wanda Kellouai, Jean-Louis Barrat, Patrick Judeinstein, Marie Plazanet, Benoit Coasne, J. Chem. Phys. 160, 024113 (2024).

Beyond well-documented confinement and surface effects arising from the large internal surface and severely confining porosity of nanoporous hosts, the transport of nanoconfined fluids remains puzzling in many aspects. With striking examples such as memory, i.e., non-viscous effects, intermittent dynamics, and surface barriers, the dynamics of fluids in nanoconfinement challenge classical formalisms (e.g., random walk, viscous/advective transport)—especially for molecular pore sizes. In this context, while molecular frameworks such as intermittent Brownian motion, free volume theory, and surface diffusion are available to describe the self-diffusion of a molecularly confined fluid, a microscopic theory for collective diffusion (i.e., permeability), which characterizes the flow induced by a thermodynamic gradient, is lacking. Here, to fill this knowledge gap, we invoke the concept of “De Gennes narrowing,” which relates the wavevector-dependent collective diffusivity D0(q) to the fluid structure factor S(q). First, using molecular simulation for a simple yet representative fluid confined in a prototypical solid (zeolite), we unravel an essential coupling between the wavevector-dependent collective diffusivity and the structural ordering imposed on the fluid by the crystalline nanoporous host. Second, despite this complex interplay with marked Bragg peaks in the fluid structure, the fluid collective dynamics is shown to be accurately described through De Gennes narrowing. Moreover, in contrast to the bulk fluid, the departure from De Gennes narrowing for the confined fluid in the macroscopic limit remains small as the fluid/solid interactions in severe confinement screen collective effects and, hence, weaken the wavevector dependence of collective transport.

Simeon Minic, Burkhard Annighofer, Arnaud Helary, Djemel Hamdane, Gaston Hui Bon Hoa, Camille Loupiac, Annie Brulet, and Sophie Combet

To probe intermediate states during unfolding and oligomerization of proteins remains a major challenge. High pressure (HP) is a powerful tool for studying these problems, revealing subtle structural changes in proteins not accessible by other means of denaturation. Bovine b-lactoglobulin (BLG), the main whey protein, has a strong propensity to bind various bioactive molecules such as retinol and resveratrol, two ligands with different affinity and binding sites. By combining in situ HPsmall- angle neutron scattering (SANS) and HP-ultraviolet/visible absorption spectroscopy, we report the specific effects of these ligands on three-dimensional conformational and local changes in BLG induced by HP. Depending on BLG concentration, two different unfolding mechanisms are observed in situ under pressures up to 300 MPa: either a complete protein unfolding, from native dimers to Gaussian chains, or a partial unfolding with oligomerization in tetramers mediated by disulfide bridges. Retinol, which has a high affinity for the BLG hydrophobic cavity, significantly stabilizes BLG both in three-dimensional and local environments by shifting the onset of protein unfolding by 100 MPa. Increasing temperature from 30 to 37C enhances the hydrophobic stabilization effects of retinol. In contrast, resveratrol, which has a low binding affinity for site(s) on the surface of the BLG, does not induce any significant effect on the structural changes of BLG due to pressure. HP treatment back and forth up to 300 MPa causes irreversible covalent oligomerization of BLG. Ab initio modeling of SANS shows that the oligomers formed from the BLG-retinol complex are smaller and more elongated compared to BLG without ligand or in the presence of resveratrol. By combining HP-SANS and HP-ultraviolet/visible absorption spectroscopy, our strategy highlights the crucial role of BLG hydrophobic cavity and opens up new possibilities for the structural determination of HP-induced protein folding intermediates and irreversible oligomerization.


Marine Le Goas, Tom Roussel, Maria Kalbazova, David Carrière, Elodie Barruet, Valerie Geertsen, Giulia C. Fadda, Fabienne Testard, Geraldine Carrot and Jean-Philippe Renault

Nanomedicines are considered as promising therapeutics for cancer treatment. However, clinical translation is still scarce, partly because their biological behavior is not well understood. Extracting general guidelines from the great variety of nanoparticles and conditions studied is indeed difficult, and relevant techniques are lacking to obtain in situ information. Here, both issues are solved by combining versatile model nanoparticles with in situ tools based on small-angle scattering techniques (SAS). The strategy was to develop a library of nanoparticles and perform systematic study of their interactions with biological systems. Considering the promising properties of gold nanoparticles as cancer therapeutics, polymethacrylate-grafted gold nanoparticles were chosen as models. Modulation of polymer chemistry was shown to change the surface properties while keeping the same structure for all nanoparticles. This unity allowed reliable comparison to extract general principles, while the synthesis versatility enabled to fine-tune the nanoparticles surface properties, especially through copolymerization, and thus to optimize their biological behavior. Two specific aspects were particularly examined: colloidal stability and cell uptake. Positive charges and hydrophobicity were identified as key parameters influencing toxicity and internalization. In situ SAS gave valuable information about nanoparticles evolution in biologically relevant environments. Good colloidal stability was thereby shown in cell culture media, while intracellular transformation and quantity of nanoparticles were monitored, highlighting the potential of these techniques for nanomedicines studies.



We show by X-ray and neutron small-angle scattering that gold nanoparticles with controlled sizes and morphologies can be obtained by the metallic reduction of AuCl4 ions trapped in 3D organic molds by X-ray radiolysis. The molds are spherical frozen micelles of polystyrene-b-poly(dimethylaminoethyl methacrylate) (PS-b-PDMAEMA) block copolymer in acidic aqueous solution with a PS spherical core surrounded by a corona of PDMAEMA chains in good solvent. The behavior of micelles is controlled by the [AuCl4]/[DMAEMA] ratio RAuCl4/DMAEMA. At low gold concentration, AuCl4 ions condense on the positively charged DMAEMA moieties without changing the behavior of the PDMAEMA chains. At intermediate gold concentration, the ions induce a progressive contraction of the corona’s chains and dehydration of micelles. At large gold concentration, the corona becomes a fully dry phase loaded with gold ions, which induces micelle aggregation. Radiolysis of the solution by an intense X-ray beam produces different types of gold nanoparticles with respect to RAuCl4/DMAEMA and irradiation time. At RAuCl4/DMAEMA = 0.033, irradiation produces in the first step gold clusters in the micelle corona which in the second step merge to form nanoparticles of a similar size to that of the micelle. Conversely, at RAuCl4/DMAEMA = 0.33, micelles do not operate as templates but only as nucleation zones and large nanoparticles grow outside the micelles.


Andrew M. Jimenez, Dan Zhao, Kyle Misquitta, Jacques Jestin and Sanat K. Kumar

Understanding the structure and dynamics of the bound polymer layer (BL) that forms on favorably interacting nanoparticles (NPs) is critical to revealing the mechanisms responsible for material property enhancements in polymer nanocomposites (PNCs). Here we use small angle neutron scattering to probe the temporal persistence of this BL in the canonical case of poly(2-vinylpyridine) (P2VP) mixed with silica NPs at two representative temperatures. We have observed almost no long-term reorganization at 150 °C (∼Tg,P2VP + 50 °C), but a notable reduction in the BL thickness at 175 °C. We believe that this apparently strong temperature dependence arises from the polyvalency of the binding of a single P2VP chain to a NP. Thus, while the adsorption–desorption process of a single segment is an activated process that occurs over a broad temperature range, the cooperative nature of requiring multiple segments to desorb converts this into a process that occurs over a seemingly narrow temperature range.



Les matériaux composites, dont on peut faire judicieusement varier la composition, permettent de combiner les propriétés de ses constituants. Ainsi, l’ajout de matériaux inorganiques (ou charge) dans une matrice polymère permet d’améliorer les propriétés d’usage de ces matériaux, telles que leurs propriétés mécaniques, électriques ou optiques, et aussi d'alléger le matériau ou d'en diminuer le coût.

Dans l’industrie du pneumatique, et en particulier dans la formulation des bandes de roulement, on utilise ainsi des matrices élastomères que l'on renforce par des particules de silice. L'étude réalisée au LLB, en collaboration avec Michelin, montrent qu'il est possible de jouer sur les effets d'entropie de mélange et la balance des contributions entropique/enthalpique des interactions, pour piloter la dispersion des particules dans la matrice et contrôler les propriétés macroscopiques des matériaux.


Des chercheurs de l’I2BC@Saclay et de l’UMR NIMBE, en collaboration avec le Laboratoire Léon Brillouin (LLB), ont analysé la structure de la couronne composée de deux protéines modèles adsorbées sur des nanoparticules de silice, en utilisant la technique de diffusion des neutrons aux petits angles. Ils montrent que les structures formées sont de véritables nanoassemblages contrôlés, dans lesquels les protéines conservent leur forme.


D. Russo, A. De Angelis, A. Paciaroni, B. Frick, N. de Sousa, F. R. Wurm, and J. Teixeira

We investigate the relaxation dynamics of proteinpolymer conjugates by neutron scattering spectroscopy to understand to which extent the coating of a protein by a polymer can replace water in promoting thermal structural fluctuations. For this purpose, we compare the dynamics of proteinpolymer mixtures to that of conjugates with a variable number of polymers covalently attached to the protein. Results show that the flexibility of the protein is larger in proteinpolymer mixtures than in native protein or in conjugates, even in the dry state. Upon hydration, both the native protein and the conjugate show equivalent dynamics, suggesting that the polymer grafted on the protein surface adsorbs all water molecules.


A. Theodoratou, L.-T. Lee, J. Oberdisse and A. Aubert-Pouëssel, Langmuir 35(20) (2019) 6620.

Abstract :

Nanofilms of about 2 nm thickness have been formed at the air–water interface using functionalized castor oil (ICO) with cross-linkable silylated groups. These hybrid films represent excellent candidates for replacing conventional polymeric materials in biomedical applications, but they need to be optimized in terms of biocompatibility, which is highly related to protein adsorption. Neutron reflectivity has been used to study the adsorption of two model proteins, bovine serum albumin and lysozyme, at the silylated oil (ICO)–water interface in the absence and presence of salt at physiologic ionic strength and pH and at different protein concentrations. These measurements are compared to adsorption at the air–water interface. While salt enhances adsorption by a similar degree at the air–water and oil–water interfaces, the impact of the oil film is significant with adsorption at the oil–water interface 3–4-fold higher compared to that at the air–water interface. Under these conditions, the concentration profiles of the adsorbed layers for both proteins indicate multilayer adsorption. The thickness of the outer layer (oil side) is close to the dimension of the minor axis of the protein molecule, ∼30 Å, suggesting a sideway orientation with the long axis parallel to the interface. The inner layer extends to 55–60 Å. Interestingly, in all cases, the composition of the oil film remains intact without significant protein penetration into the film. The optimal adsorption on these nanofilms, 1.7–2.0 mg·m-2, is comparable to the results obtained recently on thick solid cross-linked films using a quartz crystal microbalance and atomic force microscopy, showing in particular that adsorption at these ICO film interfaces under standard physiological conditions is nonspecific. These results furnish useful information toward the elaboration of vegetable oil-based nanofilms in direct nanoscale applications or as precursor films in the fabrication of thicker macroscopic films for biomedical applications.


Marguerite Léang, Didier Lairez, Fabrice Cousin, Frédérique Giorgiutti-Dauphiné, Ludovic Pauchard, Lay-Theng Lee

During evaporative drying of a colloidal dispersion, the structural behavior at the air-dispersion interface is of particular relevance to the understanding of the consolidation mechanism and the final structural and mechanical properties of the porous media. The drying-interface constitutes the region of initial drying-stress that, when accumulated over a critical thickness, leads to crack formation. This work presents an experimental study of top-down drying of colloidal silica dispersions with three different sizes (radius 5, 8 and 13 nm). Using specular neutron reflectivity, we focus on the structural evolution at the free drying-front of the dispersion with a macroscopic drying surface, and demonstrate the existence of a thick concentrated surface layer induced by heterogeneous evaporation. The reflectivity profile contains a strong structure peak due to scattering from particles in the interfacial region, from which the interparticle distance is deduced. A notable advantage of these measurements is the direct extraction of the corresponding dispersion concentration from the critical total reflection edge, providing a straightforward access to a structure-concentration relation during the drying process. The bulk reservoir of this experimental configuration renders it possible to verify the evaporation-diffusion balance to construct the surface layer and also to check reversibility of particle ordering. We follow the structural evolution of this surface layer from a sol to a soft wet-gel that is the precursor of a fragile skin, and the onset of significant particle aggregation that precedes formation of the wet-crust. Separate complementary measurements on the structural evolution in the bulk dispersion are also carried out by small angle neutron scattering, where the particle concentration is also extracted directly from the experimental curves. The two sets of data reveal similar structural evolution with concentration at the interface and in the bulk, and an increase in the degree of ordering with particle size.



Retour en haut