Cyanocobalamin-Modified Colistin-Hyaluronan Conjugates: Synthesis and Bioactivity.

Polymeric drug delivery systems enhance the biopharmaceutical properties of antibiotics by increasing their bioavailability, providing programmable and controlled-release properties, and reducing toxicity. In addition, drug delivery systems are a promising strategy to improve the intestinal permeability of various antimicrobial agents, including colistin (CT). This study describes the modification of conjugates based on CT and hyaluronic acid (HA) with cyanocobalamin (vitamin B12). Vitamin B12 was chosen as a targeting ligand because it has its own absorption pathway in the small intestine. The resulting polysaccharide conjugates contained 95 µg/mg vitamin B12 and the CT content was 335 µg/mg; they consisted of particles of two sizes, 98 and 702 nm, with a ζ-potential of approximately -25 mV. An in vitro release test at pH 7.4 and pH 5.2 showed an ultra-slow release of colistin of approximately 1% after 10 h. The modified B12 conjugates retained their antimicrobial activity at the level of pure CT (minimum inhibitory concentration was 2 µg/mL). The resulting delivery systems also reduced the nephrotoxicity of CT by 30-40% (HEK 293 cell line). In addition, the modification of B12 improved the intestinal permeability of CT, and the apparent permeability coefficient of HA-CT-B12 conjugates was 3.5 × 10-6 cm/s, corresponding to an in vivo intestinal absorption of 50-100%. Thus, vitamin-B12-modified conjugates based on CT and HA may be promising oral delivery systems with improved biopharmaceutical properties.

Chitosan Composites with Bacterial Cellulose Nanofibers Doped with Nanosized Cerium Oxide: Characterization and Cytocompatibility Evaluation.

In this work, new composite films were prepared by incorporating the disintegrated bacterial cellulose (BCd) nanofibers and cerium oxide nanoparticles into chitosan (CS) matrices. The influence of the amount of nanofillers on the structure and properties of the polymer composites and the specific features of the intermolecular interactions in the materials were determined. An increase in film stiffness was observed as a result of reinforcing the CS matrix with BCd nanofibers: the Young's modulus increased from 4.55 to 6.3 GPa with the introduction of 5% BCd. A further increase in Young's modulus of 6.7 GPa and a significant increase in film strength (22% increase in yield stress compared to the CS film) were observed when the BCd concentration was increased to 20%. The amount of nanosized ceria affected the structure of the composite, followed by a change in the hydrophilic properties and texture of the composite films. Increasing the amount of nanoceria to 8% significantly improved the biocompatibility of the films and their adhesion to the culture of mesenchymal stem cells. The obtained nanocomposite films combine a number of favorable properties (good mechanical strength in dry and swollen states, improved biocompatibility in relation to the culture of mesenchymal stem cells), which allows us to recommend them for use as a matrix material for the culture of mesenchymal stem cells and wound dressings.

Association of κ-carrageenan subjected to deep alkaline hydrolysis.

κ-carrageenan is a linear sulfated anionic gelling polysaccharide obtained from red seaweed algae by an alkaline hydrolysis. We applied static and dynamic light scattering (DLS), capillary viscometry, FT-IR, and electrophoretic DLS to gain insight into the effect of deep alkaline hydrolysis (95 °C, pH = 10 during Th = 60, 75, and 90 min) on κ-carrageenan macromolecules in a coil conformation. As DLS of coil-like κ-carrageenans is usually complicated by spurious permanent aggregates, the alkaline hydrolysis of κ-carrageenans has not been studied by DLS. By applying a double-dialysis procedure (first against water, then against 0.1 M NaCl), we succeeded in obtaining dilute solutions of coil-like κ-carrageenans with unimodal DLS distributions. Association that started with increasing concentration was attributed to attractive dipole-dipole interactions between ion-pairs formed by condensed counter-ions in the vicinity of carboxyl groups of the reducing ends. The second type of stickers resulted from the compositional heterogeneity of κ-carrageenan, namely, from the traces of ι-carrageenan blocks which amount increased with hydrolysis. With increasing Th we observed both the progressive decrease of molecular mass and the growth of association. Lowering the temperature and increasing concentration during lyophilization strengthened the intermolecular junctions, leading to the formation of permanent aggregates.

Luminescence of Eu3+ ions in hybrid polymer-inorganic composites based on poly(methyl methacrylate) and zirconia nanoparticles.

Spherical nanoparticles of ZrO2 with 2 and 10 mol% EuO1.5 up to 20 nm size were prepared by the method of hydrothermal synthesis for luminescent functionalization of the polymer-inorganic nanocomposites based on poly(methyl methacrylate). Surface modification of oxide nanoparticles was carried out by 3-(trimethoxysilyl)propyl methacrylate, dimethoxymethylvinyl silane and 2-hydroxyethyl methacrylate to provide uniform distribution and to prevent agglomeration of nanosized filler in the polymer matrix. Polymer-inorganic composites were synthesized by in situ free radical polymerization in bulk. Structuring of ZrO2 -EuO1.5 nanoparticles in the poly(methyl methacrylate) was studied by very-small-angle neutron scattering. According to the results, the dependence of photoluminescent properties of ZrO2 -EuO1.5 nanoparticles on the content of lanthanide, the symmetry of the crystal field, surface treatment and the polymer matrix were established. A correlation was shown between Stark splitting in luminescence spectra of ZrO2 -EuO1.5 nanoparticles and their phase composition. Using MMT-assay it was shown that composites based on poly(methyl methacrylate) and ZrO2 -EuO1.5 nanoparticles do not have cytotoxic properties, which makes it possible to use them as prosthesis materials with contrasted and luminescent imaging properties.

Hyaluronan/B12-chitosan polyelectrolyte complex for oral colistin administration.

Polyelectrolyte complexes (PECs) based on polysaccharides, including hyaluronic acid (HA) and chitosan (CS), are promising delivery systems for antimicrobial agents, including oral administration of the peptide antibiotic colistin (CT). Modification of CS with different targeting ligands to improve intestinal permeability is a suitable way to improve the oral bioavailability of polyelectrolyte particles. This study describes the procedure for obtaining CT-containing PECs based on HA and CS modified with cyanocobalamin (vitamin B12). In this case, vitamin B12 is used as a targeting ligand because it is absorbed in the ileum via specific transporter proteins. The resulting PECs had a hydrodynamic size of about 284 nm and a positive ζ-potential of about 26 mV; the encapsulation efficiency was 88.2 % and the CT content was 42.2 µg/mg. The developed systems provided a two-phase drug release: about 50 % of the CT was released in 0.5-1 h, and about 60 % of the antibiotic was cumulatively released in 5 h. The antimicrobial activity of encapsulated CT was maintained at the same level as the pure drug for at least 24 h (minimum inhibitory concentration against Pseudomonas aeruginosa was 2 µg/mL for both). In addition, the apparent permeability coefficient of CT in the PEC formulation was 2.4 × 10-6 cm/s. Thus, the incorporation of CT into HA- and vitamin B12-modified CS-based PECs can be considered as a simple and convenient method to improve the oral delivery of CT.

Effect of Hydroxyl-Containing Fragments on the Structure and Properties of Membrane-Forming Polyamide-Imides.

The structural features and thermophysical and transport properties of dense nonporous membranes of the casting type from (co)polyamide-imides synthesized by the polycondensation of the diacid chloride of 2-(4-carboxyphenyl)-1,3-dioxoisoindoline-5-carboxylic acid and diamines 5,5'-methylene-bis (2-aminophenol) (DADHyDPhM) and 4,4'-methylenebis(benzeneamine) (DADPhM), taken in molar ratios of 7:3, 1:1, and 3:7, have been studied. The effect of hydroxyl-containing modifying fragments of dihydroxy diphenylmethane introduced in various amounts into the main polymer chain on the pervaporation properties of the formed films is discussed. It has been shown that the presence of the residual solvent N-methyl-2-pyrrolidone in the films not only has a plasticizing effect on the characteristics of film membranes but also promotes the preferential transmembrane transport of polar liquids, primarily methanol (permeation rate over 2 kg for a copolymer with a ratio of DADHyDPhM:DADPhM = 7:3). The removal of the residual solvent from the polymer film, both thermally (heating to 200 °C) and by displacement with another solvent as a result of sequential pervaporation, led to a significant decrease in the rate of transfer of polar liquids and a decrease in the selectivity of the membrane. However, the dehydrocyclization reaction resulted in more brittle films with low permeability to penetrants of different polarities. The results of our comprehensive study made it possible to assume the decisive influence of structural changes in membranes occurring in connection with the competitive formation of intra- and intermolecular hydrogen bonds.

Synergistic Effect of Metal Oxide and Carbon Nanoparticles on the Thermal and Mechanical Properties of Polyimide Composite Films.

In this paper, we report on novel polyimide (PI) nanocomposites filled with binary mixtures of metal oxide (either TiO2 or ZrO2) nanoparticles and nanocarbon (either carbon nanofibers (CNFs) or functionalized carbon nanotubes (CNTfs)). The structure and morphology of the materials obtained were comprehensively studied. An exhaustive investigation of their thermal and mechanical properties was performed. We revealed a synergistic effect of the nanoconstituents with regard to a number of functional characteristics of the PIs compared with single-filler nanocomposites, including thermal stability, stiffness (below and above glass transition temperature), yield point, and temperature of flowing. Moreover, the possibility of manipulating the properties of the materials by choosing a proper combination of the nanofillers was demonstrated. The results obtained can become a platform in the design of PI-based engineering materials with tailored characteristics capable of operating in extreme conditions.

Metal Oxide Nanoparticles: An Effective Tool to Modify the Functional Properties of Thermally Stable Polyimide Films.

A series of polyimide/metal oxide (either ZrO2 or TiO2) nanocomposite films were fabricated based on two polymer matrices. The prepared films were characterized by scanning electron microscopy (SEM), atomic force microscopy (AFM), and X-ray diffraction analysis (XRD), and their thermal and mechanical properties were investigated with the use of thermogravimetric (TGA), differential thermal analysis (DTA), and thermomechanical analysis (TMA). We have found out that functional properties of the obtained materials are determined by a number of factors, not only the type, size, surface functionality, and concentration of the nanofiller, but also the chemical structure of the matrix polyimide. We have demonstrated some trends in the thermal and mechanical behavior of the materials depending on these features. The data could be of great interest in the areas where new materials with improved functional characteristics are needed.

Effect of Phase Heterogeneity on the Properties of Poly(vinyl alcohol)-Based Composite Pervaporation Membranes.

The structure, thermophysical characteristics, and pervaporation properties of composite membranes based on poly(vinyl alcohol) (PVA) are studied in dependence of the film preparation conditions. It is shown that the nature of the supramolecular organization of the composite polymer film determines which of the components of the separated mixtures of toluene and heptane predominantly penetrate through the corresponding pervaporation membrane. The observed structural effects can become more pronounced if the second component of a polymer mixture is purposefully selected (in this case, poly(N,N-dimethylaminoethyl methacrylate) instead of poly(acrylic acid)) or a nano-sized filler that can be well dispersed in the polymer matrix is introduced. Multi-wall carbon nanotubes are introduced into binary PVA-containing polymer blends. The influence of these fillers on the structure and transport properties of the obtained membranes is studied.

New Membrane-Forming Aromatic Co-Poly(amide-imide)s: Influence of the Chemical Structure on the Morphological, Thermal and Transport Properties.

Polymer film membranes are used to solve specific separation problems that dictate structural requirements. Structural and morphological parameters of film membranes based on glassy polyheteroarylenes can be controlled in the process of preparation from solutions that opens up prospects for obtaining structured membranes required for targeted separation. In the case of aromatic poly(amide-imide)s, the possibility of controlling film formation and structure virtually has not been studied. In the present work, a series of homologous co-poly(amide-imide)s differing in the number of repeating units with carboxyl-substituted aromatic fragments was synthesized by polycondensation. Comparative analysis of the processes of formation of membranes with different morphologies based on these polymers under equal conditions was performed. New information was obtained about the influence of the amounts of carboxyl groups and the residual solvent on structural properties of asymmetric membranes. The influence of these factors on transport properties of dense membranes under pervaporation conditions was studied. It was demonstrated that in the case of carboxyl-containing poly(amide-imide)s, the domains formed during film preparation had a significant effect on membrane properties.

Animal Cellulose with Hierarchical Structure Isolated from Halocynthia aurantium Tunic as the Basis for High-Performance Pressure-Resistant Nanofiltration Membrane.

The structure and transport properties of the new Cellokon-AKH membrane based on animal cellulose obtained from tunic of ascidian Halocynthia aurantium were studied. The results of scanning electron microscopy (SEM), FTIR spectroscopy, and the X-ray diffraction data revealed significant differences in the structure and morphology of upper and lower surfaces of this layered film membrane based on animal cellulose. It was shown that the membrane surface is a network of intertwined cellulose fibers, with both denser and looser areas present on the lower surface compared to the completely uniform morphology of the main part of the upper surface. The hierarchical structure of tunicin-based outgrowths evenly distributed over the upper surface was determined and analyzed. The 3D visual representation of the surface structure was performed with the surface reconstruction technique using scanning electron microscope images. A surface model was calculated from the aligned images based on the photogrammetric approach. The transport properties of samples with different prehistory with respect to ethanol, water, and their mixtures of different compositions were studied depending on the pressure. Representing an alcohol-containing gel film in its original state, as solvents are removed, the membrane transforms into a low-permeability fibrillary organized selective film. The obtained results confirmed the possibility of using Cellokon-AKH (dried form) for the filtration of substances with a molecular weight of more than 600 Da in various media. Further study of this new material will allow to get closer to understanding the structure of the studied seabed inhabitants and to use these natural resources more efficiently.

Bacterial Cellulose Composites with Polysaccharides Filled with Nanosized Cerium Oxide: Characterization and Cytocompatibility Assessment.

A new biocompatible nanocomposite film material for cell engineering and other biomedical applications has been prepared. It is based on the composition of natural polysaccharides filled with cerium oxide nanoparticles (CeONPs). The preparative procedure consists of successive impregnations of pressed bacterial cellulose (BC) with a sodium alginate (ALG) solution containing nanoparticles of citrate-stabilized cerium oxide and a chitosan (CS) solution. The presence of CeONPs in the polysaccharide composite matrix and the interaction of the nanoparticles with the polymer, confirmed by IR spectroscopy, change the network architecture of the composite. This leads to noticeable changes in a number of properties of the material in comparison with those of the matrix's polysaccharide composition, viz., an increase in mechanical stiffness, a decrease in the degree of planar orientation of BC macrochains, an increase in hydrophilicity, and the shift of the processes of thermo-oxidative destruction of the material to a low-temperature region. The latter effect is considered to be caused by the redox activity of cerium oxide (reversible transitions between the states Ce4+ and Ce3+) in thermally stimulated processes in the nanocomposite films. In the equilibrium swollen state, the material retains a mechanical strength at the level of ~2 MPa. The results of in vitro tests (cultivation of multipotent mesenchymal stem cells) have demonstrated the good biocompatibility of the BC-ALG(CeONP)-CS film as cell proliferation scaffolds.

Chitin Cryogels Prepared by Regeneration from Phosphoric Acid Solutions.

Cryogelation is a developing technique for the production of polysaccharide materials for biomedical applications. The formation of a macroporous structure during the freeze-drying of polysaccharide solutions creates biomaterials suitable for tissue engineering. Due to its availability, biocompatibility, biodegradability, and non-toxicity, chitin is a promising natural polysaccharide for the production of porous materials for tissue engineering; however, its use is limited due to the difficulty of dissolving it. This work describes the preparation of cryogels using phosphoric acid as the solvent. Compared to typical chitin solvents phosphoric acid can be easily removed from the product and recovered. The effects of chitin dissolution conditions on the structure and properties of cryogels were studied. Lightweight (ρ 0.025-0.059 g/cm3), highly porous (96-98%) chitin cryogels with various heterogeneous morphology were produced at a dissolution temperature of 20 ± 3 °C, a chitin concentration of 3-15%, and a dissolution time of 6-25 h. The crystallinity of the chitin and chitin cryogels was evaluated by 13C CP-MAS NMR spectroscopy and X-ray diffractometry. Using FTIR spectroscopy, no phosphoric acid esters were found in the chitin cryogels. The cryogels had compressive modulus E values from 118-345 kPa and specific surface areas of 0.3-0.7 m2/g. The results indicate that chitin cryogels can be promising biomaterials for tissue engineering.

Smart colloids containing ensembles of gold nanoparticles conjugated with κ-carrageenan.

We study colloids of nanobiocomposites (NBC) containing Au nanoparticles (NP) obtained by reduction of gold precursor with functional groups of κCAR. The AuNPs content was changed from 2.0 to 5.6 w/w %. Hydrolysis of κCAR during reduction of gold ions produced carboxylate groups supporting nanobioconjugation (FT-IR data) and electrostatic stabilization through increased negative ζ-potential. The self-assembly of AuNPs took place after storing NBC, as shown by UV/vis. extinction and transmission electron microscopy. Analysis of the structure-sensitive Rg/Rh ratios obtained by dynamic and static light scattering showed that the colloids have core-shell structure. The macromolecules in the shell exhibited polyelectrolyte expansion at [NaCl] and [KCl] < 0.02 M. Above 0.02 M, the difference between non-specific sodium and specific potassium ions on the shell thickness was registered. The temperature hysteresis of the hydrodynamic radius Rh was similar to that of the pristine κCAR observed previously by optical rotation. Monitoring the equilibration of light scattering intensity at each mini-quenching step of the cooling run allowed further insight into the hysteresis origin. These colloids are interesting for sensing applications because of enhanced scattering of plasmonically coupled AuNPE. Their thermo- and ion-sensitivity can be useful in drug delivery.

Pervaporation membranes of a simplex type with polyelectrolyte layers of chitosan and sodium hyaluronate.

Self-supporting multilayer films containing a polyelectrolyte complex (PEC) were prepared by the sequential layering of sodium hyaluronate (HA, MW 5.4 × 104) and chitosan (CS, MW 1.6 × 105, the degree of deacetylation 0.80) in different orders. Imaging with low-voltage scanning electron microscopy (LVSEM) showed that the CS/HA films had a multilayer structure, while X-ray diffraction (XRD) indicated significant structuring of the CS layer near the PEC-CS region. Analysis of the thermal properties of the CS/HA films revealed differences in the structural organization and morphological features of the polymer layers and high thermal stability of the PEC layer. Testing of the transport properties of the CS/HA film in pervaporation (PV) separation using different compositions of ethanol-water mixtures indicated that the multilayer membrane was selective across a wide range of concentrations in the feed. Separation of an azeotropic ethanol-water mixture containing 5 wt% water yielded a permeate consisting of about 100 wt% water. LVSEM revealed that the membrane microstructure changed during the PV process due to membrane swelling and changes in the arrangement of the macromolecules during transport of the penetrant. The results support the use of CS/HA composite films as highly effective PV membranes. In addition to pervaporation separation, CS/HA multilayer films can also be used for drug delivery, tissue engineering, and wound healing applications.

Preparation of N-succinyl-chitin nanoparticles and their applications in otoneurological pathology.

Succinyl-chitin (SCH) nanoparticles were obtained by acylation of partially deacetylated chitin (DCH) nanofibers. Introduction of the succinyl moiety induced a partial amorphization of DCH, as viewed by X-ray diffraction, and increased the fractal dimension of the colloids from df = 1.2 (DCH) to 1.5-1.7 (SCH), as revealed by light scattering. The spherically symmetric form of the colloids remained almost unchanged, as indicated by the range of structure-sensitive ratios 1.0 < Rg/Rh < 1.2; the hydrodynamic diameter ranged from 200 to 300 nm. The cytoprotective activity of the SCH nanoparticles was evaluated in vivo in an acute hearing pathology model (220-250 g male Wistar rats, n = 90) following prophylactic and therapeutic administrations. Ototropic action was estimated using the amplitude of otoacoustic emissions at the frequency of the distortion product otoacoustic emissions in the range of 4-6.4 kHz before acoustic stimulation, as well as at 1 h, 24 h, and 7 days after acoustic stimulation. A dispersion of 0.3% SCH nanoparticles demonstrated prolonged ototropic action and earlier regeneration of hearing functions when compared to a meglumine sodium succinate solution. Thus, intravenous administration of the SCH nanoparticles increases the cycling time of exogenous succinate and improves biodistribution in tissues possessing a hemato-labyrinth barrier.

O,N-(2-sulfoethyl)chitosan: Synthesis and properties of solutions and films.

A series of water-soluble sulfoethylated chitosans (SEC) with degrees of substitution (DS) up to 130% were obtained using a heterogeneous reaction of chitosan with sodium 2-chloroethanesulfonate in 85% isopropanol in the presence of NaOH. NMR and FTIR spectroscopy confirmed that sulfoethylation of chitosan preferentially happens at hydroxyl groups and to some extent at amino groups, giving mixed substituted O,N-SEC. Chitosan shows positive birefringence, whereas SEC shows negative values, indicating self-organization in dilute solution. Dynamic light scattering studies revealed the presence of aggregates in dilute solutions of chitosan and SEC. The sizes of the SEC aggregates are sensitive to the DS and the nature of the solvent. X-ray diffraction of SEC films revealed that the introduction of sulfoethyl groups into chitosan leads to amorphization, which is more pronounced at higher DS. During the storage of SEC films, the samples loose solubility due to the formation of ionic crosslinks upon dehydration.