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The adsorption of lipids onto spherical polymer colloids led to original assemblies presenting structural characteristics adjustable with the lipid formulation. The model system selected for this work involved sulfate-charged poly(styrene) submicrometer particles and zwitterionic/cationic lipid mixtures composed of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-dipalmitoyl-3-trimethylammonium-propane (DPTAP). According to the theoretical packing parameter calculations and whatever the DPPC/DPTAP ratio, the two lipids self-assembled in aqueous media to spontaneously form vesicles. A phase transition investigation of these DPPC/DPTAP vesicles using differential scanning calorimetry revealed particular thermotropic behaviors, especially for the equimolar formulation where very strong interactions occurred between DPPC and DPTAP. Furthermore, the coating of the lipids around particles was monitored versus DPPC/DPTAP ratio by means of numerous appropriate techniques. First, a thermogravimetric analysis, providing decomposition profiles of lipid/polymer particle assemblies with temperature, was atypically carried out for such nanostructures. Then, ¹H NMR spectroscopy enabled the exact DPPC/DPTAP molar ratios adsorbed on particles to be determined by differentiating both lipids. Subsequently, it also pointed out the major role of electrostatic interactions as driving forces in the assembly elaboration process. In addition to these findings, quantitative information has been collected and correlated with chemical lipid assays and permitted the statement of a lipid bilayer coverage for the assemblies prepared in water, in agreement with quasi-elastic light scattering data.

Abstract: Using the stepwise heterocoagulation concept, fluorescent and highly magnetic submicronic core-shell polymer particles were prepared. For this purpose a negatively charged oil-in-water magnetic emulsion was first modified by adsorbing the poly(ethyleneimine) (PEI). Secondly, low glass transition temperature (T g=10°C) anionic film-forming nanoparticles were adsorbed onto the cationic magnetic droplets. Finally the encapsulation was induced by heating the heterocoagulates above the T g of the film-forming nanoparticles. To produce labeled magnetic particles, fluorescent nanoparticles and film-forming nanoparticles were simultaneously adsorbed. PEI adsorption was investigated. Also investigated was the influence of the amount of film-forming nanoparticles and fluorescent nanoparticles on the encapsulation efficiency.

This work investigated the elaboration of biocompatible nanoparticles from the pH-induced self-complexation of the amphoteric polysaccharide N-sulfated chitosan. The acidification of aqueous solutions of chitosan having a degree of acetylation of 24% and a degree of sulfation of 34% or 56% was followed stepwise by turbidimetry, dynamic light scattering, and electrophoresis. With the highest sulfated chitosan, no turbidity was recorded between pH = 7.8 and 2.0, traducing a high apparent solubility of the polymer chains in this domain of pH. With the lowest sulfated chitosan, a steady increase in turbidity was monitored from pH = 6.90 to 6.15 followed by the flocculation of the polymer at pH ~ 6.0. In this range of pH, the polymer phase separated to yield particles having hydrodynamic diameters decreasing from 350 to 260 nm and an almost constant negative charge. These particles were assembled by electrostatic interactions between the protonated amino residues and the sulfate functions and stabilized by an excess of surface sulfate groups. The particles could be separated from the reaction medium and concentrated by centrifugation−redispersion cycles without alteration of their structure.

An amphiphilic block copolymer poly(tert-butylacrylamide-b-(N-acryloylmorpholine-N-acryloxysuccinimide)) (poly(TBAm-b-(NAM/NAS)) and a random copolymer poly(NAM/NAS), synthesized by the reversible addition−fragmentation chain transfer (RAFT) polymerization process, have been used as support for oligonucleotide (ODN) synthesis, to elaborate polymer−oligonucleotide conjugates. In a first step, starters of ODN solid-phase synthesis were coupled to activated ester functions of polymers, and second, resulting functionalized polymers were covalently grafted onto hydroxylated controlled pore glass (CPG) support to further accomplish ODN synthesis. An efficient capping of residual hydroxyl functions of CPG was performed before synthesis, with both acetic anhydride and diethoxy-N,N-diisopropyl-phosphoramidite reagents, to suppress parasite-free ODN population present in conjugate crude material and resulting from syntheses directly initiated on silica beads. After purification, conjugates were evaluated in a DNA hybridization assay on a microarray, as macromolecules being able to favor capture of the target. Conjugate coating conditions were studied on the dT25/dA25 model. The role of the hydrophobic part (poly(TBAm)) of the conjugate synthesized with the block copolymer in the orientation of the conjugate after coating was revealed by spotting experiments achieved in a mixed solvent (DMF/H2O). The use of block copolymer−dT25 conjugate afforded a significant sensitivity improvement of the hybridization assay.

A model study was investigated to develop colloidal supramolecular assemblies consisting of particles coated with lipid layers. The interactions between monodisperse sulfate-charged poly(styrene) submicrometer particles and zwitterionic/cationic lipid vesicles composed of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine and 1,2-dipalmitoyl-3-trimethylammonium propane were considered. The influence of relevant experimental parameters on the final associations was examined by quasi-elastic light scattering to point out some new phenomena occurring in these colloidal systems. The major role of electrostatic interactions as driving forces to control the organization between cationic lipids and oppositely charged poly(styrene) particles was clearly evident, whereas this influence was less pronounced when considering the zwitterionic lipids. The characterization of these original complex assemblies was completed by a thorough study of the surface modification. The combination of zeta potential measurements, X-ray photoelectron spectroscopy analyses, and microscopy observations proved that the envisioned model can really correspond to polymer particles surrounded by lipids.

The formation of colloids based on polyelectrolyte complexes (PECs) of biopolymers was investigated through the complexation between two charged polysaccharides, chitosan as polycation, and dextran sulfate as polyanion. The slow dropwise addition of components, generally used for the formation of PECs, allowed to elaborate both cationic or anionic particles with an excess of chitosan or dextran sulfate, respectively. The PEC particles featured a core/shell structure, the hydrophobic core resulting from the segregation of complexed segments whereas excess component in the outer shell ensured the colloidal stabilization against further coagulation. Considering the host/guest concept for the formation of PECs, the influence of the molecular weight of components on particles sizes could be well explained by the chain length ratios of the two polymers. As an irreversible flocculation occurred with a dropwise approach for both cationic and anionic PEC particles when the mixing ratio was close to unity, a more versatile, and simpler to setup, method was designed:  the one-shot addition of one solution to the other. Because process of addition is faster than the flocculation, cationic or anionic particles could be elaborated irrespective of the order of addition of the reactant. Characterization of these particles by quasielastic light scattering, electrophoresis, and scanning electron microscopy revealed very similar properties to those obtained by a slow dropwise approach. Critical coagulation concentrations of 0.12 and 0.09 M (with sodium chloride) for cationic and anionic particles evidenced a mostly electrostatic stabilization.

Formation of colloids based on polyelectrolyte complexes (PECs) was mainly studied with synthetic polyelectrolytes. In this study, we describe the elaboration of positively charged PEC particles at a submicrometer level obtained by the complexation between two charged polysaccharides, chitosan as polycation and dextran sulfate (DS) as polyanion. The complexes were elaborated by dropwise addition of default amounts of DS to excess chitosan. Quasi-elastic light scattering was used to investigate in detail the influence of the characteristics of components (chain length, degree of acetylation) and parameters linked to the reaction of complexation (molar mixing ratio, ionic strength, concentration in polymer) on the sizes and polydispersity of colloids. Chain length of chitosan is the major parameter affecting the dimensions of the complexes, high molar mass chitosans leading to the largest particles. Variations of hydrodynamic diameters of PECs with the molar mass of chitosan are consistent with a mechanism of particle formation through the segregation of the neutral and then hydrophobic blocks of the polyelectrolyte complexed segments. Resulting particles display probably a structure constituted by a neutral core surrounded by a chitosan shell ensuring the colloidal stabilization. Such a structure was evidenced by measurements of electrophoretic mobilities revealing that the positive charge of particles was decreasing with pH, in relation with the neutralization of excess glucosamine hydrochloride moieties.

The adsorption of a rhodamine X labeled oligonucleotide composed of 25-mers of thymine (dT25-ROX) onto the thermosensitive shell of PMMA/PNIPAM core−shell latex particles was studied at 22 and 40 °C, below and above the TVPT (volume phase transition temperature) of the PNIPAM shell, respectively. The experimental binding isotherms were well fitted with the cooperative Hill model. The Hill coefficient is lower than 1 at both temperatures showing that the adsorption is anticooperative. The polarity of the shell was probed by both the lifetimes and solvatochromic shifts of the zwitterionic form of rhodamine X. For temperatures below the shell TVPT has a polarity similar to that of water, while for temperatures above the transition the polarity is equivalent to that of a water/dioxane mixture with 30% (v/v) water.

A matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI−TOF MS) study of N-acryloylmorpholine (NAM) homopolymers, as obtained by reversible addition−fragmentation chain transfer (RAFT) polymerization technique, is reported. Polymerization of NAM was performed in dioxane using azobis(isobutyronitrile) (AIBN) as initiator and tert-butyl dithiobenzoate as RAFT chain transfer agent. Polymer samples of low molecular weights (<10 000 g mol^-1) and low polydispersity indices (PDI < 1.1) were obtained, which are essential requirements for such MALDI−TOF MS analysis. First, analysis of poly(NAM) samples in linear mode led to %@mt;sys@%%@ob@%%@ital@%M%@rsf@%%@sb@%n%@sbx@%%@obx@%%@mx@% values very close to both size exclusion chromatography/light scattering (SEC/LS) and ¹H NMR values as well as to the theoretical ones. Then, an accurate examination of chain end groups was performed using the reflectron mode. Two main chain populations were identified:  (i) dormant chains (i.e., initiated by a tert-butyl and terminated by a dithiobenzoate group) together with sulfine and thioester-ended chains probably resulting from oxidation of dithiobenzoate chain ends during storage; (ii) proton-terminated chains mainly produced by fragmentation of the former chains in the spectrometer. In addition, some chains which could correspond to termination reactions onto the intermediate radicals involved in the RAFT equilibrium were suspected. Finally, comparison of polymer samples before and after aminolysis indicated that dithioester-ended chains constituted the majority of chains initially present in these samples. This study confirms that it is indeed possible to use MALDI−TOF MS to investigate the structure of polymer chains synthesized by the RAFT technique.

The neutralization of a homogeneous series of chitosans with degrees of acetylation (DA) varying between 1% and 71% was followed stepwise by static light scattering. The determination of the gyration radii (RG,z) and the second virial coefficients (A2) allowed us a better understanding of the chitosan behavior during the neutralization in aqueous solutions. On increasing pH, a critical value was evidenced both by a drop of RG,z and a fall of A2. The values of the critical pH were also shown as highly dependent on the polymer concentration and the ionic strength of the media. On increasing DA, the range of pH where chitosan was apparently soluble enlarged although the second virial coefficient decreased with both the increase of DA and the degree of neutralization. Working at the critical pH, just before precipitation occurred, transmission electronic microscopy allowed us to evidence a sol/gel transition responsible for the formation of a stable colloidal dispersion. The average particle sizes increased with the degree of acetylation:  from 100 nm for DA = 1% to 300 nm for DA = 51%. These aggregates were probably assembled both by hydrophobic interactions and hydrogen bonding due to the neutralization of the ammonium groups into −NH2.

Nano size-ranged poly(d,l-lactic acid) (PLA) particles were obtained from 2 and 10 w% PLA solution in water-saturated ethyl acetate by the emulsification-diffusion method. An increase in the PLA solution concentration resulted in an increase of the particle mean size from ca 260 nm to 530 nm with a broadening of the distribution. After fractionation by centrifugation, the particle mean size was 980 nm. The Pluronic F68 used in the formulations at concentration ranging from 0.5 to 5% w/v, was shown to adsorb at the interface at 0.5 mg/m 2 for both particle sizes. The molecular occupied area of 24 nm 2/molecule suggested an expanded conformation of the surfactant at the interface, though an incomplete coverage could not be excluded. Coagulation kinetics measurements revealed a critical coagulation concentration (CCC) of 1.5 M sodium chloride, indicating a steric stabilization of the colloids by the adsorbed triblock copolymer. Nonetheless, variations in zeta potential with increasing salt concentration were observed, suggesting that the surface carboxylate groups were still accessible for further modifications.