Marguerite Rinaudo

Recent studies on semi-dilute Calf-Thymus DNA (CT-DNA) solutions have revealed the presence of birefringence and small-scale textures influenced by shear and ionic strength. In this study, we investigate these phenomena on the same solutions to elucidate the underlying shear-induced supramolecular organization and relaxation dynamics using rheo-birefringence, rheology, and rheo-SAXS (small-angle X-ray scattering). Static SAXS confirmed concentration-dependent inter-chain correlations in the 15–25 nm range, while rheology revealed a slipping yield-stress behavior. Oscillatory strain sweep and steady state rheo-birefringence experiments correlated the appearance of textures with the onset of flow and a stress plateau observed over a shear rate range from approximately 1 to 1000 s⁻¹. Transient rheo-birefringence and rheo-SAXS revealed two distinct relaxation mechanisms on well-separated time scales: a fast process lasting a few seconds, inversely proportional to the shear rate, consistent with the orientational relaxation of DNA segments on a ~20 nm scale; and a slower relaxation over tens of seconds, independent of the shear rate, associated with the disappearance of textures, and attributed to a diffusive process. These findings provide significant insights into the mechanisms governing DNA organization and dynamics in semi-dilute solutions under flow and highlight the need for temporally resolved start-up rheo-SAXS and rheo-birefringence measurements, as well as theoretical models describing these processes across various spatial and temporal scales.

Agarose forms a homogeneous thermoreversible gel in an aqueous solvent above a critical polymer concentration. Contrary to the prevailing consensus, recent confirmations indicate that agarose gels are also stable in non-solvents like acetone and ethanol. A previous study compared gel characterisations and behaviours in water and ethanol, discussing the gelation mechanism. In the current work, the ethanol gel is exchanged with water to explore the potential reversibility of the displacement of water in agarose. Initially, the structure is characterised using ¹H NMR in DMSO-d₆ and D₂O solvents. Subsequently, a very low yield (0.04) of methyl substitution per agarobiose unit is determined. The different gels after stabilisation are characterised using rheology, and their physical properties are compared based on the solvent used. The bound water molecules, acting as plasticizers in aqueous medium, are likely removed during the exchange process with ethanol, resulting in a stronger and more fragile gel. Next, the gel obtained after the second exchange from ethanol back to water is compared with the initial gel prepared in water. This is the first time where such gel has been characterised without undergoing a phase transition when switching from a good solvent to a non-solvent, and vice versa, thereby testing the reversibility of the solvent exchange. Reversibility of this behaviour is demonstrated through swelling and rheology experiments. This study extends the application of agarose in chromatography and electrophoresis.

Agarose is known to form a homogeneous thermoreversible gel in an aqueous medium over a critical polymer concentration. The solid-liquid phase transitions are thermoreversible but depend on the molecular structure of the agarose sample tested. The literature has mentioned that agarose gels could remain stable in non-solvents such as acetone or ethanol. However, there has been no characterization of their behavior nor a comparison with the gels formed in a good solvent such as water. In the first step of this article, the structure was characterized using ¹H and ¹³C NMR in both D₂O and DMSO-d6 solvents. DMSO is a solvent that dissolves agarose regardless of the temperature. First, we have determined a low yield of methyl substitution on the D-galactose unit. Then, the evolution of the ¹H NMR spectrum was monitored as a function of temperature during both increasing and decreasing temperature processes, ranging from 25 to 80 °C. A large thermal hysteresis was obtained and discussed, which aided in the interpretation of rheological behavior. The hysteresis of NMR signals is related to the mobility of the agarose chains, which follows the sol/gel transition depending on the chains’ association with H-bonds between water and the -OH groups of agarose for tightly bound water and agarose/agarose in chain packing. In the second step of the study, the water in the agarose gel was exchanged with ethanol, which is a non-solvent for agarose. The resulting gel was stable, and its properties were characterized using rheology and compared to its behavior in aqueous media. The bound water molecules that act as plasticizers were likely removed during the exchange process, resulting in a stronger and more brittle gel in ethanol, with higher thermal stability compared to the aqueous gel. It is the first time that such gel is characterized without phase transition when passing from a good solvent to a non-solvent. This extends the domains of application of agarose.

Agarose is known to form a homogeneous thermoreversible gel in aqueous medium over a critical polymer concentration. The solid-liquid phase transitions are thermoreversible but depend on the molecular structure of the agarose sample tested. Then, in a first step, the structure was characterised by 1H and 13C NMR in D2O and in DMSO which is a solvent of agarose whatever the temperature. A low yield in methyl substituent on the D-galactose unit was determined. Then, evolution of the 1H NMR spectrum was followed as a function of temperature in increasing and decreasing temperature process from 25 to 80°C. A large thermal hysteresis is obtained and discussed. It helps to interpret rheological behaviour. In fact, NMR signals are related to proton relaxation and especially to proton involved in H-bonds between water and -OH agarose for tightly bound water and agarose/agarose in chain packing. In a second step, water was exchanged against ethanol which is a non solvent of agarose. A stable gel was demonstrated and characterised by rheology to be compared with aqueous behaviour. Bound water playing the role of plasticizer is probably removed and the gel is much stronger (and brittle) in ethanol with a larger thermal stability. It is the first time that such gel is characterised without phase transition when passing from a good-solvent to a non-solvent. This extends the domains of application of agarose.

In this work, purified pectins from Araçá fruits (Psidium cattleianum Sabine) were obtained and characterized after partial demethylation. On each prepared sample, the carboxylic yield was obtained by titration, the degree of methylation (DM) by ¹H-NMR, and the molecular weight distribution by steric exclusion chromatography (SEC). Then, the gelation ability in the presence of calcium counterions was investigated and related to DM (59–0%); the pectin concentration (2–10 g L⁻¹); and the CaCl₂ concentration (0.1–1 mol L⁻¹) used for dialysis. The critical pectin concentration for homogeneous gelation was above 2 g L⁻¹ when formed against 1 mol L⁻¹ CaCl₂. The elastic modulus (G′) increased with pectin concentration following the relationship G′~C^2.8 in agreement with rigid physical gel network predictions. The purified samples APP and APP-A with DM ≥ 40% in the same conditions released heterogeneous systems formed of large aggregates. Gels formed against lower concentrations of CaCl₂ down to 0.1 mol L⁻¹ had a higher degree of swelling, indicating electrostatic repulsions between charged chains, thus, counterbalancing the Ca²⁺ cross-linkage. Compression/traction experiments demonstrated that an irreversible change in the gel structure occurred during small compression with an enhancement of the G′ modulus.

Christian Enrique Garcia Garcia, Frédéric Bossard , Bernard Lardy and Marguerite Rinaudo This paper describes the development of our work on electrospinning of chitosan (CS) to get pure chitosan nanofiber mats proposed as support for chondrocyte culture.

Our objective in this work was to summarize the main results obtained in processing pure chitosan and chitosan/hyaluronan complex in view of biomedical applications, taking advantage of their original properties. In addition, an electrospinning technique was selected to prepare nanofiber mats well adapted for tissue engineering in relation to the large porosity of the materials, allowing an exchange with the environment. The optimum conditions for preparation of purified and stable nanofibers in aqueous solution and phosphate buffer pH = 7.4 are described. Their mechanical properties and degree of swelling are given. Then, the prepared biomaterials are investigated to test their advantage for chondrocyte development after comparison of nanofiber mats and uniform films. For that purpose, the adhesion of cells is studied by atomic force microscopy (AFM) using single-cell force spectroscopy, showing the good adhesion of chondrocytes on chitosan. At the end, adhesion and proliferation of chondrocytes in vitro are examined and clearly show the interest of chitosan nanofiber mats compared to chitosan film for potential application in tissue engineering.

In this work, optimized conditions for preparation of chitosan and hyaluronan polyelectrolyte complex are proposed. The objective was to produce new biomaterials being biocompatible and bioresorbable in the body as well as approaching the extracellular matrix (ECM) structure. These materials will be tested for chondrocyte development in tissue engineering and wound healing applications. Nanofibers made of the polyelectrolyte complex (PEC) were successfully manufactured by electrospinning, and casted films were used as a model for properties comparison. To our knowledge, it is the first time that stable chitosan/hyaluronan fibers are produced, which were observed to be long-lasting in buffer at pH~7.4. The role of thermal treatment at 120 °C for 4 h is examined to control the degree of swelling by crosslinking of the two polysaccharides by H-bonds and amide bonds formation. The properties of the materials are tested for different PEC compositions at different pH values, based on swelling and solubility degrees, diameters of nanofibers and mechanical performances. The influence of the solvent (acidic potential and composition) utilized to process biomaterials is also examined. Acid formic/water 50/50 v/v is observed to be the more appropriated solvent for the carried-out procedures.