Electrospun Composites of Chitosan with Cerium Oxide Nanoparticles for Wound Healing Applications: Characterization and Biocompatibility Evaluation In Vitro and In Vivo.

Cerium oxide nanoparticles (CeONPs), as part of tissue regeneration matrices, can protect cells from reactive oxygen species and oxidative stress. In addition, they can influence the properties of the scaffold, including its electrospinnability and mechanical strength. In this work, we prepared electrospun fiber mats from a chitosan and polyethylene oxide blend (CS-PEO) with the addition of ceria nanoparticles (CS-PEO-CeONP). The addition of CeONPs resulted in a smaller fiber diameter and higher swelling compared to CS-PEO fiber mats. CeONP-modified fiber mats also had a higher Young's modulus due to the reinforcing effect of the nanoparticles. Both mats had comparable adhesion and cytocompatibility to mesenchymal stem cells, which had a more rounded morphology on CS-PEO-CeONP compared to elongated cells on the CS-PEO mats. Biocompatibility in an in vivo rat model showed no acute toxicity, no septic or allergic inflammation, and no rough scar tissue formation. The degradation of both mats passed the stage of matrix swelling. CS-PEO-CeONP showed significantly slower biodegradation, with most of the matrix remaining in the tissue after 90 days. The reactive inflammation was aseptic in nature with the involvement of multinucleated foreign-body type giant cells and was significantly reduced by day 90. CeONPs induced the formation of the implant's connective tissue capsule. Thus, the introduction of CeONPs influenced the physicochemical properties and biological activity of CS-PEO nanofiber mats.

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.

Biocomposite films based on chitosan and cerium oxide nanoparticles with promising regenerative potential.

Polymeric nanocomposite materials have great potential in the development of tissue-engineered scaffolds because they affect the structure and properties of polymeric materials and regulate cell proliferation and differentiation. In this work, cerium oxide nanoparticles (CeONPs) were incorporated into a chitosan (CS) film to improve the proliferation of multipotent mesenchymal stem cells (MSCs). The citrate-stabilized CeONPs with a negative ζ-potential (-25.0 mV) were precoated with CS to obtain positively charged particles (+20.3 mV) and to prevent their aggregation in the composite solution. The composite CS-CeONP films were prepared in the salt and basic forms using a dry-cast process. The films obtained in both forms were characterized by a uniform distribution of CeONPs. The incorporation of CeONPs into the salt form of CS increased the stiffness of the CS-CeONP film, while the subsequent conversion of the film to the basic form resulted in a decrease in both the Young's modulus and the yield stress. The redox activity (Ce4+ ⇌ Ce3+) of cerium oxide in the CS-CeONP film was confirmed by thermal oxidative degradation. In vitro culture of MSCs showed that the CS-CeONP film has good biocompatibility, and in vivo experiments demonstrated its substantial regenerative potential.

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.

Biophysical Characterization and Cytocompatibility of Cellulose Cryogels Reinforced with Chitin Nanowhiskers.

Polysaccharide-based cryogels are promising materials for producing scaffolds in tissue engineering. In this work, we obtained ultralight (0.046-0.162 g/cm3) and highly porous (88.2-96.7%) cryogels with a complex hierarchical morphology by dissolving cellulose in phosphoric acid, with subsequent regeneration and freeze-drying. The effect of the cellulose dissolution temperature on phosphoric acid and the effect of the freezing time of cellulose hydrogels on the structure and properties of the obtained cryogels were studied. It has been shown that prolonged freezing leads to the formation of denser and stronger cryogels with a network structure. The incorporation of chitin nanowhiskers led to a threefold increase in the strength of the cellulose cryogels. The X-ray diffraction method showed that the regenerated cellulose was mostly amorphous, with a crystallinity of 26.8-28.4% in the structure of cellulose II. Cellulose cryogels with chitin nanowhiskers demonstrated better biocompatibility with mesenchymal stem cells compared to the normal cellulose cryogels.

Bacterial Cellulose (Komagataeibacter rhaeticus) Biocomposites and Their Cytocompatibility.

A series of novel polysaccharide-based biocomposites was obtained by impregnation of bacterial cellulose produced by Komagataeibacter rhaeticus (BC) with the solutions of negatively charged polysaccharides-hyaluronan (HA), sodium alginate (ALG), or κ-carrageenan (CAR)-and subsequently with positively charged chitosan (CS). The penetration of the polysaccharide solutions into the BC network and their interaction to form a polyelectrolyte complex changed the architecture of the BC network. The structure, morphology, and properties of the biocomposites depended on the type of impregnated anionic polysaccharides, and those polysaccharides in turn determined the nature of the interaction with CS. The porosity and swelling of the composites increased in the order: BC-ALG-CS > BC-HA-CS > BC-CAR-CS. The composites show higher biocompatibility with mesenchymal stem cells than the original BC sample, with the BC-ALG-CS composite showing the best characteristics.

Cytocompatibility of Bilayer Scaffolds Electrospun from Chitosan/Alginate-Chitin Nanowhiskers.

In this work, a bilayer chitosan/sodium alginate scaffold was prepared via a needleless electrospinning technique. The layer of sodium alginate was electrospun over the layer of chitosan. The introduction of partially deacetylated chitin nanowhiskers (CNW) stabilized the electrospinning and increased the spinnability of the sodium alginate solution. A CNW concentration of 7.5% provided optimal solution viscosity and structurization due to electrostatic interactions and the formation of a polyelectrolyte complex. This allowed electrospinning of defectless alginate nanofibers with an average diameter of 200-300 nm. The overall porosity of the bilayer scaffold was slightly lower than that of a chitosan monolayer, while the average pore size of up to 2 µm was larger for the bilayer scaffold. This high porosity promoted mesenchymal stem cell proliferation. The cells formed spherical colonies on the chitosan nanofibers, but formed flatter colonies and monolayers on alginate nanofibers. The fabricated chitosan/sodium alginate bilayer material was deemed promising for tissue engineering applications.

Electrospun Bilayer Chitosan/Hyaluronan Material and Its Compatibility with Mesenchymal Stem Cells.

A bilayer nonwoven material for tissue regeneration was prepared from chitosan (CS) and hyaluronic acid (HA) by needleless electrospinning wherein 10-15 wt% (with respect to polysaccharide) polyethylene oxide was added as spinning starter. A fiber morphology study confirmed the material's uniform defect-free structure. The roughness of the bilayer material was in the range of 1.5-3 µm, which is favorable for cell growth. Electrospinning resulted in the higher orientation of the polymer structure compared with that of corresponding films, and this finding may be related to the orientation of the polymer chains during the spinning process. These structural changes increased the intermolecular interactions. Thus, despite a high swelling degree of 1.4-2.8 g/g, the bilayer matrix maintained its shape due to the large quantity of polyelectrolyte contacts between the chains of oppositely charged polymers. The porosity of the bilayer CS-HA nonwoven material was twice lower, while the Young's modulus and break stress were twice higher than that of a CS monolayer scaffold. Therefore, during the electrospinning of the second layer, HA may have penetrated into the pores of the CS layer, thereby increasing the polyelectrolyte contacts between the two polymers. The bilayer CS-HA scaffold exhibited good compatibility with mesenchymal stem cells. This characteristic makes the developed material promising for tissue engineering applications.

Modification of the Ceramic Implant Surfaces from Zirconia by the Magnetron Sputtering of Different Calcium Phosphate Targets: A Comparative Study.

In this study, thin calcium phosphate (Ca-P) coatings were deposited on zirconia substrates by radiofrequency (RF) magnetron sputtering using different calcium phosphate targets (calcium phosphate tribasic (CPT), hydroxyapatite (HA), calcium phosphate monobasic, calcium phosphate dibasic dehydrate (DCPD) and calcium pyrophosphate (CPP) powders). The sputtering of calcium phosphate monobasic and DCPD powders was carried out without an inert gas in the self-sustaining plasma mode. The physico-chemical, mechanical and biological properties of the coatings were investigated. Cell adhesion on the coatings was examined using mesenchymal stem cells (MSCs). The CPT coating exhibited the best cell adherence among all the samples, including the uncoated zirconia substrate. The cells were spread uniformly over the surfaces of all samples.