Laurent Veron

A Novel Synthesis of Chitosan Nanoparticles in Reverse Emulsion

Physical hydrogels of chitosan in the colloidal domain were obtained in the absence of both cross-linker and toxic organic solvent. The approach was based on a reverse emulsion of a chitosan solution in a Miglyol/Span 80 mixture, generally regarded as safe. Temperature and surfactant concentration were optimized, and the impact of the degree of acetylation (DA) and the molar mass of chitosan was investigated. When chitosan had a DA above 30%, only macroscopic gels were obtained, because of the predominance of attractive Van der Waals forces. The lower the molar mass of chitosan, the better the control over particle size and size distribution, probably as a result of either a reduction in the viscosity of the internal aqueous phase or an increase in the disentanglement of the polymer chain during the process. After extraction and redispersion of the colloid in an ammonium acetate buffer, the composition of the particles was around 80% of pure chitosan corresponding to a recovery of 60% of the original input. These new and safe colloids offer wide perspectives of development in further applications.

Synthesis and structural characterization of chitosan nanogels.

Colloidal physical gels of pure chitosan were obtained via an ammonia-induced gelation in a reverse phase emulsion. The water weight fraction and the chitosan concentration in the water phase were optimized so as to yield nanogels with controlled particle size and size distribution. The spherical morphology of the nanogels was established by transmission electron microscopy with negative staining. Wide-angle X-ray scattering experiments showed that these gels were partially crystalline. The electrophoretic mobilities of the particles remained positive up to pH 7, above which the particles aggregated due to the charge neutralization. From the investigation on the colloidal stability of these nanogels in various conditions (pH, salt concentration, temperature), an electrosteric stabilization process of the particles was pointed out, related to the conformation of mobile chitosan chains at the gel-liquid interface. Therefore, the structure of the nanogels was deduced as being core-shell type, a gelified core of neutralized chitosan chains surrounded by partially protonated chains.

Rapid Bacterial Identification, Resistance, Virulence and Type Profiling using Selected Reaction Monitoring Mass Spectrometry

Mass spectrometry (MS) in Selected Reaction Monitoring (SRM) mode is proposed for in-depth characterisation of microorganisms in a multiplexed analysis. Within 60–80 minutes, the SRM method performs microbial identification (I), antibiotic-resistance detection (R), virulence assessment (V) and it provides epidemiological typing information (T). This SRM application is illustrated by the analysis of the human pathogen Staphylococcus aureus, demonstrating its promise for rapid characterisation of bacteria from positive blood cultures of sepsis patients.

Self‐Assemblies on Chitosan Nanohydrogels

Nanohydrogels of pure chitosan, containing neither potentially toxic solvent nor chemical cross-linker, were obtained by an ammonia-induced physical gelation of a reverse emulsion of a chitosan solution in a triglyceride mixture as an organic phase. The resulting colloids were obtained with a controlled size distribution and displayed a positive surface charge. Assemblies with various macromolecules were investigated as a first step toward new nano-carriers for bioactive molecules. Chondroitin sulfate formed polyelectrolyte complexes with the positively charged surface of the nanogels, leading to negative chitosan-based colloidal hydrogels with preservation of the original average size of the dispersion. The mode of assembly of HIV-1 p24 protein with these colloids relied on multiple interactions between the protein and the hydrogels, irrespective of their surface charges. Anyhow, the amounts of loaded protein remained limited, suggesting a surface association. The assembly of an immunoglobulin (IgG) was markedly different from p24. No association was detected with the positive colloidal hydrogels whereas a very high loading capacity could be obtained with the negative ones. So, this work reports that fully biodegradable submicrometric physical hydrogels could be obtained from naturally occurring polymers. These gels could cargo a variety of biomolecules making them versatile carriers with many potential applications in Life Sciences.