M.T. Di Bari

Structure of self-organized multilayer nanoparticles for drug delivery.

The combined use of cryo-TEM, dynamic light scattering, and small-angle X-ray and neutron scattering techniques allows a detailed structural model of complex pharmaceutical preparations of soybean lecithin/chitosan nanoparticles used as drug vectors to be worked out. Charge-driven self-organization of the lipid(-)/polysaccharide(+) vesicles occurs during rapid injection, under mechanical stirring, of an ethanol solution of soybean lecithin into a chitosan aqueous solution. We conclude that beyond the charge inversion region of the phase diagram, i.e., entering the redissolution region, the initial stages of particle formation are likely to be affected by a re-entrant condensation effect at the nanoscale. This behavior resembles that at the mesoscale which is well-known for polyion/amphiphile systems. Close to the boundary of the charge inversion region, nanoparticle formation occurs under a maximum condensation condition at the nanoscale and the complexation-aggregation process is driven toward a maximum multilamellarity. Interestingly, the formulation that maximizes vesicle multilamellarity corresponds to that displaying the highest drug loading efficiency.

IN13 Backscattering Spectrometer at ILL: Looking for Motions in Biological Macromolecules and Organisms

In 1998, three partner groups (the French institutions Institut de Biologie Structurale and the Léon Brillouin Laboratory and the Italian Istituto Nazionale per la Fisica della Materia, now merged with the Consiglio Nazionale delle Ricerche, INFM-CNR) applied to operate the thermal backscattering spectrometer IN13, at the Institut Laue Langevin, as a French-Italian Collaborative Research Group (CRG). The plan was to have access to a dedicated spectrometer in order to explore how far neutron scattering could contribute to the understanding of dynamics in biological macromolecules: how “flexible” must be a biological object to perform its function?

Structure and organization of phospholipid/polysaccharide nanoparticles

In recent years nanoparticles and microparticles composed of polymeric or lipid material have been proposed as drug carriers for improving the efficacy of encapsulated drugs. For the production of these systems different materials have been proposed, among them phospholipids and polysaccharides due to their biocompatibility, biodegradability, low cost and safety. We report here a morphological and structural investigation, performed using cryo-TEM, static light scattering and small angle neutron and x-ray scattering, on phospholipid/saccharide nanoparticles loaded with a lipophilic positively charged drug (tamoxifen citrate) used in breast cancer therapy. The lipid component was soybean lecithin; the saccharide one was chitosan that usually acts as an outer coating increasing vesicle stability. The microscopy and scattering data indicate the presence of two distinct nanoparticle families: uni-lamellar vesicles with average radius 90 A and multi-lamellar vesicles with average radius 440 A. In both families the inner core is occupied by the solvent. The presence of tamoxifen gives rise to a multi-lamellar structure of the lipid outer shell. It also induces a positive surface charge into the vesicles, repelling the positively charged chitosan molecules which therefore do not take part in nanoparticle formation.

Elastic incoherent neutron scattering as a probe of high pressure induced changes in protein flexibility.

We report here the results of elastic incoherent neutron scattering experiments on three globular proteins (trypsin, lysozyme and beta-lactoglobulin) in different pressure intervals ranging from 1 bar to 5.5 kbar. A decrease of the mean square hydrogen fluctuations, u(2), has been observed upon increasing pressure. Trypsin and beta-lactoglobulin behave similarly while lysozyme shows much larger changes in u(2). This can be related to different steps in the denaturing processes and to the high propensity of lysozyme to form amyloids. Elastic incoherent neutron scattering has proven to be an effective microscopic technique for the investigation of pressure induced changes in protein flexibility.

Dynamics of hydrated starch saccharides

We report here elastic neutron scattering data on glucose and on two of its polymeric forms: amylose and amylopectin. We have covered the hydration range from the dry state to about 0.6 g water/g dry saccharide. The data indicate, in all the analysed systems, the presence of a dynamic glass-like transition similar to that observed in hydrated proteins. The fact that this feature is observed also in a relatively small molecule like glucose confirms the hypothesis already put forward by other authors, that this transition in biomolecular species is essentially triggered and driven by the interaction of the macromolecule with the network of fluctuating H-bond of the solvent.

Pressure dependence of protein dynamics investigated using elastic and quasielastic neutron scattering

We present here a study of the dynamics of two monomeric proteins, trypsin and lysozyme, by means of elastic and quasielastic neutron scattering under medium-to-high pressure conditions (1-1200 bar). The internal motions as probed from the average proton dynamics in the 100 ps timescale have a confined diffusive nature. On increasing the pressure up to 1200 bar the confinement volume is almost unaffected, while the fraction of protons involved is slightly decreased.

FINSLER GEOMETRIC LOCAL INDICATOR OF CHAOS FOR SINGLE ORBITS IN THE HENON-HEILES HAMILTONIAN

Translating the dynamics of the Henon--Heiles hamiltonian as a geodesic flow on a Finsler manifold, we obtain a local and synthetic Geometric Indicator of Chaos (GIC) for two degrees of freedom continuous dynamical systems. It represents a link between local quantities and asymptotic behaviour of orbits giving a strikingly evident, one-to-one, correspondence between geometry and instability.

Dynamics of trypsin under pressure

Abstract The pressure dependence of the dynamics of a monomeric protein has been studied by quasielastic neutron scattering. The EISF shows a reduction in the volume sampled by the protons in going from 1 to 900 atm. A slight decrease of the quasielastic broadening with pressure is also observed.

QENS investigation of the dynamics of starch saccharides

We present here quasi-elastic neutron scattering results on D20 hydrated samples of amylose, one of the main saccharide components of starch. Two different sample hydrations (h = 0.5 and 1.0 g D2O (g dry amylose)-1 have been investigated in the temperature range 170 to 350 K. Below 260 K only an elastic contribution is present in the spectra, while a quasi-elastic component shows up above this temperature. The elastic incoherent structure factor (EISF) associated with this component changes considerably with increasing temperature. For the sample with hydration h = 0.5 the confinement volume increases by a factor of four in going from 300 to 350 K, and the proportion of hydrogen involved in the confined diffusion motion increases as well from 30 to 55%. Similar effects are observed at the higher hydration investigated. The observed dynamics can be associated with the known plasticising role of water in polysaccharides.

Dynamics of hydrated saccharides of interest in food science

Abstract We have used IN13 at the ILL to investigate the dynamics of amylose and amylopectine performing elastic scans from 20 to 320 K at different hydration levels. In the dry samples, the atomic dynamics is harmonic in the whole temperature range explored. For the hydrated samples the mean square atomic displacement, 〈u2〉, is almost hydration independent and harmonic up to ∼230 K for amylose, and ∼250 K for amylopectine. A ‘glass-like’ kinetic transition then shows up with a marked increase in 〈u2〉: this behaviour is qualitatively similar to that observed in other biopolymers like globular proteins, but here the transition temperature is higher and it markedly varies with hydration.