Elaboration of nano-organized assemblies of "lipids/polysaccharide particles" for biomedical applications

Une nouvelle génération d’assemblages nano-organisés, constitués de couches lipidiques enveloppant des cœurs particulaires sphériques, a été développée ces dernières années. Un des nombreux intérêts de cette génération d’assemblages, à structure bio-inspirée, est de constituer in fine des vecteurs ou réservoirs pour des applications biomédicales de délivrance contrôlée de principes actifs. Ce type d’architecture originale est obtenu par réorganisation de membranes lipidiques à la surface de nanoparticules de polymère. Au cours de ce travail, la sélection du support colloïdal s’est portée sur des nanoparticules de chitosane, un biopolymère aux propriétés physico-chimiques et biologiques remarquables. Ces nanoparticules ont été élaborées par une méthode dite de « gélification ionique » à l’aide de tripolyphosphate de sodium. L’optimisation de ce procédé de synthèse a permis l’obtention de nanoparticules cationiques et sphériques, de taille reproductible d’une centaine de nanomètres, et de distribution en taille étroite. Aucune modification de ces caractéristiques de taille n'a été montrée durant au moins 4 mois à 23°C dans l'eau. Quant aux membranes lipidiques, il a été choisi d’élaborer des vésicules ou des nano-disques modèles et anioniques de façon à présenter une charge électrostatique opposée à celle des surfaces particulaires. La caractérisation de ces entités en termes de taille, distribution en taille, charge de surface et morphologie a été effectuée par diffusion quasi-élastique de la lumière, microscopie électronique à transmission, diffusion de neutrons aux petits angles et zêtamétrie. L’influence de différents paramètres physico-chimiques impliqués dans la formation des assemblages (i.e., force ionique et pH de la phase continue, rapport lipides/nanoparticules) a été examinée, permettant d’une part, une meilleure compréhension des interactions s’établissant entre les chaînes de chitosane et les membranes lipidiques, et d’autre part, l’obtention d’assemblages de taille et de distribution en taille satisfaisantes. Une étude de l’incorporation de différentes molécules thérapeutiques (isoniazide, ibuprofène et rose bengale) dans le support particulaire a ensuite été réalisée afin d’évaluer le potentiel des nanoparticules de chitosane, puis des assemblages lipidiques en tant que vecteurs de principes actifs. Des études préliminaires ont ainsi pu mettre en évidence que ce recouvrement surfacique lipidique apportait une modification satisfaisante du profil de relargage du principe actif.A novel generation of nano-organized assemblies, composed of lipid layers surrounding spherical nanoparticle cores, has been recently developed for drug delivery applications. This original architecture results of the lipid membrane reorganization onto colloidal polymer surfaces. In the present study, the colloidal supports are constituted of chitosan nano-hydrogels, a biopolymer with exceptional physicochemical and biological properties. These colloidal supports have been elaborated via an ionic gelation process using sodium tripolyphosphate as gelation agent. The optimization of all the parameters involved in this process has led to cationic spherical nanoparticles, with reproducible sizes in nanometer range, and narrow size distributions. No alteration of these characteristics has been observed for at least 4 months in water at 23 °C. Regarding the lipid membranes, two different morphologies (i.e., vesicle and nanodisc) have been prepared. Lipid formulation has been designed to obtain objects with a negative surface charge, opposed to chitosan nanoparticle one. These entities were characterized in terms of size, size distribution, surface charge and morphology using dynamic light scattering, transmission electron microscopy as well as small angle neutron scattering. The assembly of colloidal supports and lipid membranes has been investigated by studying the influence of the different physicochemical parameters involved in the synthesis. During this study, phenomena involved in the adsorption process were pointed out, and experimental conditions were optimized in order to control the assembly elaboration process. Finally, a study on drug incorporation (isoniazid, ibuprofen and rose bengale) into chitosan nanoparticles has been carried out. Preliminary experiments on drug release from nanoparticles and assemblies revealed a promising effect of the surface modification of assemblies on their drug release profile

Interests of chitosan nanoparticles conically cross-linked with tripolyphosphate for biomedical applications

International audienceThe process of ionic gelation is one of the easiest ways to develop chitosan nanoparticles reported so far in the literature. Its success is mainly due to its one-shot synthesis, and to the mild environment required to produce the nanoparticles. The nanoparticle formation all along this process has been therefore thoroughly studied to lead to particles with a nanometric size, a narrow size distribution, and a spherical shape that are ideal for biomedical uses. The purpose of this review is to compile the biomedical applications that have been considered in the literature for these chitosan nanoparticles prepared by ionic gelation using tripolyphosphate as ionic agent. Their intrinsic biological properties such as non-toxicity, antimicrobial activity, mucoadhesivity and haemocompatibility are firstly discussed and compared to those of chitosan solutions. Then, the different bioactive species (drugs and biomacromolecules) incorporated in these chitosan nanoparticles, their maximal incorporation efficiency, their loading capacity, and their principal associated biomedical applications are presented

A close collaboration of chitosan with lipid colloidal carriers for drug delivery applications

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Elaboration of chitosan nanoparticles: Favorable impact of a mild thermal treatment to obtain finely divided, spherical, and colloidally stable objects

International audienceThe elaboration of chitosan (CS) nanoparticles via an ionic gelation process using sodium tripolyphosphate (TPP) as cross-linking agent was thoroughly studied in order to develop colloidally stable, spherical CS nanoparticles with a reproducible sub-micrometer size, and a narrow size distribution. To this end, the most relevant parameters involved in the synthesis such as CS chains concentration and molecular weight as well as ionic strength and pH of CS initial solutions were investigated and optimized. The physicochemical characterization of resulting CS nanoparticles was carried out in terms of size, size distribution, and surface charge by quasi-elastic light scattering (QELS), nanoparticle tracking analysis (NTA), and zeta potential measurements. Morphological characterization using classical and cryogenic transmission electron microscopy (TEM and cryo-TEM) was then performed revealing a high number of aggregates mixed to individualized nanoparticles with drop-like and irregular shapes. These limitations were overcome by applying a specific and optimized thermal treatment which appeared to significantly reduce the aggregate number, and to restructure the CS nanoparticles into highly spherical objects. Based on all these findings, reproducible, cationic (zeta potential = +37 +/- 5 mV), stable (at least 4 months at 20 degrees C), spherical, and mainly individualized CS nanoparticles of 100 +/- 30 nm (determined from cryo-TEM images) were obtained

Tunable morphology of lipid/chitosan particle assemblies

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Chitosan Films for Microfluidic Studies of Single Bacteria and Perspectives for Antibiotic Susceptibility Testing

International audienceSingle-cell microfluidics is a powerful method to study bacteria and determine their susceptibility to antibiotic treatment. Glass treatment by adhesive molecules is a potential solution to immobilize bacterial cells and perform microscopy, but traditional cationic polymers such as polylysine deeply affect bacterial physiology. In this work, we chemically characterized a class of chitosan polymers for their biocompatibility when adsorbed to glass. Chitosan chains of known length and composition allowed growth of Escherichia coli cells without any deleterious effects on cell physiology. Combined with a machine learning approach, this method could measure the antibiotic susceptibility of a diversity of clinical strains in less than 1 h and with higher accuracy than current methods. Finally, chitosan polymers also supported growth of Klebsiella pneumoniae, another bacterial pathogen of clinical significance. IMPORTANCE Current microfluidic techniques are powerful to study bacteria and determine their response to antibiotic treatment, but they are currently limited by their complex manipulation. Chitosan films are fully biocompatible and could thus be a viable replacement for existing commercial devices that currently use polyly-sine. Thus, the low cost of chitosan slides and their simple implementation make them highly versatile for research as well as clinical use

Small anticancer drug release by light: Photochemical internalization of porphyrin-β-cyclodextrin nanoparticles

International audienceAiming to engineer simple, neutral, strongly amphiphilic photoactive nanoparticles (NPs) to specifically target cancer cell lysosomes for drug transport and light-controlled release, new conjugates of β-cyclodextrin with highly hydrophobic triphenylporphyrin bearing different alkyl chains, were synthesized. Although differently sized, all conjugates self-assemble into ~60 nm NPs in water and display similar photoactivity. The NPs target selectively the lysosomes of breast adenocarcinoma MCF-7 cells, embedding in vesicular membranes, as experiments with model liposomes indicate. Either empty or drug-loaded, the NPs lack dark toxicity for 48 h. They bind with differently structured anticancer drugs tamoxifen and gemcitabine as its N-adamantyl derivative. Red light irradiation of cells incubated with drug-loaded NPs results in major reduction of viability (>85 %) for 48 h displaying significant synergy of photo-chemotoxicity, as opposed to empty NPs, and to loaded non-irradiated NPs, in manifestation of photochemical internalization (PCI). Our approach expands the field of PCI into different small molecule chemotherapeutics