Structure, the energy, sorption and biological properties of chiral salts of chitosan with l- and d-ascorbic acid.

The influence of l- and d-ascorbic acid diastereomers on the structure, supramolecular ordering, energy, sorption and biological properties of heterochiral (D-L) and homochiral (D-D) salt complexes of chitosan (d-glucan)-acid was studied. The thermal effect of dissolving chitosan in l-ascorbic acid and the protonation degree of (D-L)-salts were lower than those in the medium of the d-isomer. Homochiral (D-D) salts, in contrast to heterochiral (D-L) ones, are distinguished by a more developed system of intermolecular and intramolecular contacts, a more ordered and equilibrium supramolecular organization of macrochains, a higher crystallinity degree, and a smaller amount of crystallization water. The sorption isotherms of chiral salts were approximated by the thermal equation of sorption and the superposition of the Langmuir and Flory-Huggins isotherms. Significant differences were found in the limiting value and energy of sorption, the constant of adsorption equilibrium, the limiting sorption capacity of the localized mode of water, and the Gibbs mixing energy. Biotesting on non-vascular (Scenedesmus quadricauda) and vascular eukaryotes (Linum usitatissimum) revealed the growth-stimulating effect of the D-D salts. The obtained results confirm our hypothesis of the homochiral salt complexes d-glucan-d-ascorbic acid best corresponding to the principles of the functional organization of biological objects.

Structural and Morphological Features of Anisotropic Chitosan Hydrogels Obtained by Ion-Induced Neutralization in a Triethanolamine Medium.

For the first time, anisotropic hydrogel material with a highly oriented structure was obtained by the chemical reaction of polymer-analogous transformation of chitosan glycolate-chitosan base using triethanolamine (TEA) as a neutralizing reagent. Tangential bands or concentric rings, depending on the reaction conditions, represent the structural anisotropy of the hydrogel. The formation kinetics and the ratio of the positions of these periodic structures are described by the Liesegang regularities. Detailed information about the bands is given (formation time, coordinate, width, height, and formation rate). The supramolecular ordering anisotropy of the resulting material was evaluated both by the number of Liesegang bands (up to 16) and by the average values of the TEA diffusion coefficient ((15-153) × 10-10 and (4-33) × 10-10 m2/s), corresponding to the initial and final phase of the experiment, respectively. The minimum chitosan concentration required to form a spatial gel network and, accordingly, a layered anisotropic structure was estimated as 1.5 g/dL. Morphological features of the structural anisotropic ordering of chitosan Liesegang structures are visualized by scanning electron microscopy. The hemocompatibility of the material obtained was tested, and its high sorption-desorption properties were evaluated using the example of loading-release of cholecalciferol (loading degree ~35-45%, 100% desorption within 25-28 h), which was observed for a hydrophobic substance inside a chitosan-based material for the first time.

Thermosensitive Chitosan-Containing Hydrogels: Their Formation, Properties, Antibacterial Activity, and Veterinary Usage.

Mixtures of aqueous solutions of chitosan hydrochloride (CS·HCl, 1-4 wt.%) and Pluronic F-127 (Pl F-127, 25 wt.%) were studied using vibrational and rotational viscometry; the optimal aminopolysaccharide concentration (3 wt.%) and the CS·HCl:Pl F-127 ratio (30:70) to obtain a thermosensitive hydrogel were found. It was shown that at 4 °C, such mixed compositions were viscous liquids, while at 37 °C for 1-2 min, they undergo a thermally reversible transition to a shape-stable hydrogel with a developed level of structure formation, satisfactory viscosity and high mucoadhesive parameters (maximum pull-off force Fmax = 1.5 kN/m2; work of adhesion W = 66.6 × 10-3 J). Adding D-ascorbic acid to the hydrogel led to orientational ordering of the supramolecular structure of the mixed system and significantly improved mucoadhesion (Fmax = 4.1 kN/m2, W = 145.1 × 10-3 J). A microbiological study revealed the high antibacterial activity of the hydrogel against Gram-negative and Gram-positive bacterial strains. The treatment of mixed bacterial infection in cows demonstrated the possibility of the in situ formation of a viscoelastic gel and revealed its high therapeutic effect. It has been suggested that our thermosensitive mucoadhesive CS·HCl:Pl F-127 hydrogels could be considered as independent veterinary drugs and pharmaceuticals.

Formation of Liesegang Structures under the Conditions of the Spatiotemporal Reaction of Polymer-Analogous Transformation (Salt → Base) of Chitosan.

This paper presents the results of our study of the diffusion front progression of OH- ions in the bulk of a flat layer of the salt chitosan form during the course of the chemical reaction of the polymer-analogous salt → base transformation, accompanied by the formation of a water-insoluble polymeric phase as a gel film with its structure of concentric rings or tangential bands. Using scanning electron microscopy and polarization microscopy, structural and morphological features of these periodic formations were visualized. Physicochemical parameters and spatiotemporal mass transfer characteristics were obtained. The process under study is shown to obey the classical laws of ion-exchange reactions, and the formation kinetics and the ratio of the positions of periodic structures are described by the laws characteristic of the Liesegang phenomenon. The average diffusion coefficients of hydroxide ions in the periodic formations calculated, considering the protonation degree of amino groups, were found to be no more than a decimal order of magnitude lower than those in an aqueous solution. This confirms the fact of swelling of the formed chitosan base and the existence of a gel film, in whose liquid phase diffusion occurs.

Optical activity anisotropy of chitosan-based films.

The optical activity [α] of films of chitosan and chitosan acetate was studied. [α] depended on the orientation angle θ of the film sample around the light beam. The angular dependences [α] = f(θ) (indicatrices) were processed to extract the constant term [α]0and four harmonics ([α]i, Fourier's series) determined by structure elements with the corresponding symmetry: the amorphous (isotropic) phase of chitosan ([α]0), irregular-shaped structures ([α]1), rod-shaped ones in the film plane ([α]2), helical ones located perpendicular to the film surface ([α]3), and crystalline structures ([α]4). The contribution of individual supramolecular structures to the overall optical activity of chitosan was estimated: [α]2 and [α]4 predominated for the fresh chitosan acetate films, while [α]4 did for the chitosan ones. The results of the numerical decomposition of indicatrices are in satisfactory agreement with X-ray diffractometry data. The observed regularities predetermine prospects for obtaining stereoselective chiro-optic chitosan-based materials with novel functional properties.

Influence of Chitosan Ascorbate Chirality on the Gelation Kinetics and Properties of Silicon-Chitosan-Containing Glycerohydrogels.

The influence of the chirality of chitosan ascorbate on the gelation kinetics and the properties of hybrid silicon-chitosan-containing glycerohydrogels were studied with a deep estimation of the stereospecificity of chitosan polysalts with l- and d-ascorbic acid diastereomers and their biological effects. It has been established that l- and d-diastereomerically enriched chitosan ascorbates are characterized by a positive Cotton effect and differ in the wavelength of the maximum of the dichroic band (250 and 240 nm), as well as in the values of its specific ellipticity (21.8 × 105 and 39.2 × 105 deg·mL·dm-1·g-1), the sign of specific optical rotation (+ and -), the type of dispersion curves (anomalous and smooth), as well as the condensed phase morphology (anisodiametric particles with optical anisotropy and confocal domains of spherical shape, respectively). In the biomimetic sol-gel synthesis of silicon-chitosan-containing glycerohydrogels using silicon tetraglycerolate as a precursor, it was found that chitosan d-ascorbate retarded gelation. Thin congruent plates obtained from the corresponding glycerohydrogels based on chitosan d-ascorbate have higher mechanical strength and elasticity under uniaxial stretching and lower values of Young's modulus. It has been shown that the systems based on chitosan d-ascorbate show the greatest antibacterial activity against Staphylococcus aureus 209P and Escherichia coli 113-13 and significantly promote the viability of normal human dermal fibroblasts. The results of our assessment of the biological properties of chitosan polysalts are unexpected, since ascorbic acid exhibits biological activity as its l-isomer only.