Porous Polylactide Microparticles as Effective Fillers for Hydrogels.

High-strength composite hydrogels based on collagen or chitosan-genipin were obtained via mixing using highly porous polylactide (PLA) microparticles with diameters of 50-75 µm and porosity values of over 98%. The elastic modulus of hydrogels depended on the filler concentration. The modulus increased from 80 kPa to 400-600 kPa at a concentration of porous particles of 12-15 wt.% and up to 1.8 MPa at a filling of 20-25 wt.% for collagen hydrogels. The elastic modulus of the chitosan-genipin hydrogel increases from 75 kPa to 900 kPa at a fraction of particles of 20 wt.%. These elastic modulus values cover a range of strength properties from connective tissue to cartilage tissue. It is important to note that the increase in strength in this case is accompanied by a decrease in the density of the material, that is, an increase in porosity. PLA particles were loaded with C-phycocyanin and showed an advanced release profile up to 48 h. Thus, composite hydrogels mimic the structure, biomechanics and release of biomolecules in the tissues of a living organism.

Chitosan Sponges for Efficient Accumulation and Controlled Release of C-Phycocyanin.

The paper proposed a new porous material for wound healing based on chitosan and C-phycocyanin (C-PC). In this work, C-PC was extracted from the cyanobacteria Arthrospira platensis biomass and purified through ammonium sulfate precipitation. The obtained C-PC with a purity index (PI) of 3.36 ± 0.24 was loaded into a chitosan sponge from aqueous solutions of various concentrations (250, 500, and 1000 mg/L). According to the FTIR study, chitosan did not form new bonds with C-PC, but acted as a carrier. The encapsulation efficiency value exceeded 90%, and the maximum loading capacity was 172.67 ± 0.47 mg/g. The release of C-PC from the polymer matrix into the saline medium was estimated, and it was found 50% of C-PC was released in the first hour and the maximum concentration was reached in 5-7 h after the sponge immersion. The PI of the released C-PC was 3.79 and 4.43 depending on the concentration of the initial solution.

Biomechanical behaviour of PEDOT:PSS-based hydrogels as an electrode for stent integrated enzyme biofuel cells.

The possibility of creating a biofuel cell based on a metal stent was shown in this study. Given the existing stent implantation approaches, the integration of a biofuel cell into a stent naturally entails capacity for biofuel cells to be installed into a human body. As a counter electrode, a hydrogel based on iota-carrageenan, polyvinyl alcohol, and PEDOT:PSS, with an immobilized glucose oxidase enzyme, was proposed. Tension tests demonstrated that the hydrogel mechanical behavior resembles that of a bovine's vein. To obtain an analytical description, the deformation curves were fitted using Gent and Ogden models, prompting the fitting parameters which can be useful in further investigations. During cyclic biaxial studies the samples strength was shown to decreases insignificantly in the first 50 cycles and, further, remains stable up to more than 100 cycles. The biofuel cell was designed with the PEDOT:PSS based material as an anode and a Co-Cr self-expanding stent as a cathode. The maximum biofuel cell power density with a glucose concentration of 5 mM was 7.87 × 10-5 W in phosphate buffer and 3.98 × 10-5 W in blood mimicking buffer. Thus, the biofuel cell integration in the self-expanding stent was demonstrated.

Magnetic levitational bioassembly of 3D tissue construct in space.

Magnetic levitational bioassembly of three-dimensional (3D) tissue constructs represents a rapidly emerging scaffold- and label-free approach and alternative conceptual advance in tissue engineering. The magnetic bioassembler has been designed, developed, and certified for life space research. To the best of our knowledge, 3D tissue constructs have been biofabricated for the first time in space under microgravity from tissue spheroids consisting of human chondrocytes. Bioassembly and sequential tissue spheroid fusion presented a good agreement with developed predictive mathematical models and computer simulations. Tissue constructs demonstrated good viability and advanced stages of tissue spheroid fusion process. Thus, our data strongly suggest that scaffold-free formative biofabrication using magnetic fields is a feasible alternative to traditional scaffold-based approaches, hinting a new perspective avenue of research that could significantly advance tissue engineering. Magnetic levitational bioassembly in space can also advance space life science and space regenerative medicine.

Noninvasive high-frequency acoustic microscopy for 3D visualization of microstructure and estimation of elastic properties during hydrolytic degradation of lactide and ε-caprolactone polymers.

The monitoring of degradation processes' kinetics in polymers is one of the attractive possibilities of ultrasound technique applications that provide non-destructive imaging of polymers' internal microstructure and measurements of elastic properties. In this work, biodegradable polymers and copolymers based on L,L-lactide, D,L-lactide and ε-caprolactone have been studied at different stages of hydrolysis at 37 °C by high-frequency (100 and 200 MHz) ultrasound. The acoustic microscopy technique has been developed to reveal changes in the internal microstructure and bulk sound speed in polymer samples over a hydrolysis period of 25 weeks. Ultrasound imaging provides visualization of amorphous and crystalline phases, internal imperfections, variation in packing density, and other microstructural features. Acoustic images demonstrate nucleation, growth, and the changes in internal inhomogeneities in polymers during degradation accompanied by a decrease in the polymers' molecular weight. We associate the changes in the elastic properties (the speed of a longitudinal wave) with crystallinity variations in polymers during hydrothermal aging. The results of the ultrasound investigations are supplemented by gel permeation chromatography, differential scanning calorimetry, and wide-angle X-ray spectroscopy. STATEMENT OF SIGNIFICANCE: Monitoring the kinetics of degradation processes in polymers is one of the attractive possibilities of applying ultrasound techniques that provide non-destructive imaging of the polymers' internal microstructure and measurements of elastic properties. In this work, visualization of nucleation, growth, and evolution of internal inhomogeneities in the volume of polymers and variation of values of speed of longitudinal and transverse sound waves during hydrolysis are compared with measurements of molecular weight, density, data of DSC curves, and X-ray scattering analysis. We discuss several common phenomena that occur in the volume of poly(L-lactide) and poly(D,L-lactide) over the degradation process as well as improvement of elastic properties of the poly(ε -caprolactone) and poly(L-lactide-co-caprolactone) during hydrothermal aging.

Electroconductive PEDOT:PSS-based hydrogel prepared by freezing-thawing method.

Biopolymer-based composition with adding of conductive polymer poly-(3,4-ethylenedioxythiophene) polystyrenesulfonate (PEDOT PSS) was made by mixing of iota-carrageenan (CRG), polyvinyl alcohol (PVA) and PEDOT PSS followed by freezing/thawing cycles. The method is environmentally friendly and based on the formation of polymer matrix upon of mixing CRG, PVA and PEDOT PSS and formation of porous physical gel due to freezing/thawing cycles. It is necessary to mention that all components are well-known as biocompatible materials. The resulting material is stable in water and also has swelling capability both in distilled water and physiological solutions. Structure of material was characterized by means of X-ray diffraction, optical and electron microscopy. Electrophysical investigations also were performed. The conductivity of the gel immersed in distilled water is comparable with the dry gel value and close to 0.01 [S/cm].

Viscoll collagen solution as a novel bioink for direct 3D bioprinting.

Collagen is one of the most promising materials for 3D bioprinting because of its distinguished biocompatibility. Cell-laden constructs made of pure collagen with or without incorporated growth supplements support engineered constructs persistence in culture and are perfectly suitable for grafting. The limiting factor for direct 3D collagen printing was poor printability of collagen solutions, especially admixed with cells or tissue spheroids. In our study, we showed that concentrated solutions of native collagen branded Viscoll were effective as bioinks with high fidelity performance. Viscoll containing 20, 30, or 40 mg/ml collagen were used for direct extrusion 3D bioprinting to form scaffolds appropriate to support spatial arrangement of tissue spheroids into rigid patterns with resolution of 0.5 mm in details. Incorporated cells demonstrated sufficient viability. Associated rheological study showed that good printability of the collagen solutions correlates with their increased storage modulus value, notably exceeding the loss modulus value. The proper combination of these physical parameters could become technological criteria for manufacturing various collagen bioinks for 3D bioprinting.

In vitro assessment of electrospun polyamide-6 scaffolds for esophageal tissue engineering.

Artificial tissue-engineered grafts offer a potential alternative to autologous tissue grafts for patients, which can be traumatic. After decellularizing Papio hamadryas esophagus and studying the morphology and physical properties of the extracellular matrix (ECM), we generated electrospun polyamide-6 based scaffolds to mimic it. The scaffolds supported a greater mechanical load than the native ECM and demonstrated similar 3D microstructure, with randomly aligned fibers, 90% porosity, 29 µm maximal pore size, and average fiber diameter of 2.87 ± 0.95 µm. Biocompatibility studies showed that human adipose- and bone marrow-derived mesenchymal stromal cells (AD-MSC and BMD-MSC) adhered to the scaffold surface and showed some proliferation: scaffold cell coverage was 25% after 72 h of incubation when seeded with 1000 cells/mm2 ; cells elongated processes along the polyamide-6, although they flattened 1.67-4 times less than on cell culture plastic. Human umbilical vein endothelial cells, however, showed poor adherence and proliferation. We thus provide in vitro evidence that polyamide-6 scaffolds approximating the esophageal biomechanics and 3D topography of nonhuman primates may provide a biocompatible substrate for both AD-MSC and BMD-MSCs, supporting their adhesion and survival to some degree. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 107B: 253-268, 2019.

Multi-hierarchical tissue-engineering ECM-like scaffolds based on cellulose acetate with collagen and chitosan fillers.

A novel high-tech composite biomimetic matrixes for a wide range of medical purposes were prepared. The structure of scaffolds was inspired by the architecture of native decellularized tissue: material consists of a sponge and fibrous components of different spatial geometry based on cellulose acetate with collagen or chitosan filler. The fibrous component was prepared by electrospinning, the sponge - freeze-drying technique. The influence of main technological parameters, such as freeze mode, polymer type and concentration, etc. on the fiber-sponge architecture and properties was examined. It was shown that scaffolds with different types of microstructure can be obtained employing this technique. The impregnation of chitosan or collagen filler in fiber matrix also significantly improves mechanical properties up to 40 MPa for strength and 600 MPa for Young's modulus.

Collagen tissue treated with chitosan solution in H2O/CO2 mixtures: Influence of clathrates hydrates on the structure and mechanical properties.

A mixture of water/carbon dioxide is a "green" perspective solvent from the viewpoint of biomedical applications. Clathrate hydrates are formed this solvent under certain conditions and a very interesting question is the impact of clathrates hydrates on the structure and properties of bovine pericardium, which is used in biomedicine, in particular as a main part of biological heart valve prostheses. The aim of the present work is to investigate the influence of clathrates on the structure and mechanical properties of the collagen tissue treated with chitosan in H2O/CO2 mixtures under pressure 3.0-3.5MPa and temperatures 2-4°C. It was first found that the clathrate hydrates in this media due to the strong fluctuations "bomb" collagen tissue of bovine pericardium, which is manifested in the appearance of numerous small gaps (pores) with mean size of 225±25nm and large pores with size of 1-3µ on the surface and within collagen matrices. High porosity leads to averaging characteristics of the organization structure in tissues with different orientation of the collagen fibers. As a result, the mechanical properties of the collagen tissue with a different orientation of the collagen fibrils become similar, which is quite different from their original properties. The structural changes caused by the influence of the environment clathrate hydrates led to a significant decrease of the tensile strength (30-47% in total, p<0.05) and initial elastic moduli (74-83%, p<0.05). However, the final elastic moduli and the maximum tensile virtually unchanged compared to the control. Nevertheless, it was found that the direct deposition of chitosan from the H2O/CO2 mixtures with clathrate improve the mechanical-strength properties of the porous matrices. We believe that these improved mechanical properties are achieved due to particularly deep and uniform impregnation of the collagen matrix with chitosan from its pressurized solutions in H2O/CO2 mixtures.

Orthotopic transplantation of a tissue engineered diaphragm in rats.

The currently available surgical options to repair the diaphragm are associated with significant risks of defect recurrence, lack of growth potential and restored functionality. A tissue engineered diaphragm has the potential to improve surgical outcomes for patients with congenital or acquired disorders. Here we show that decellularized diaphragmatic tissue reseeded with bone marrow mesenchymal stromal cells (BM-MSCs) facilitates in situ regeneration of functional tissue. A novel bioreactor, using simultaneous perfusion and agitation, was used to rapidly decellularize rat diaphragms. The scaffolds retained architecture and mechanical properties and supported cell adhesion, proliferation and differentiation. Biocompatibility was further confirmed in vitro and in vivo. We replaced 80% of the left hemidiaphragm with reseeded diaphragmatic scaffolds. After three weeks, transplanted animals gained 32% weight, showed myography, spirometry parameters, and histological evaluations similar to native rats. In conclusion, our study suggested that reseeded decellularized diaphragmatic tissue appears to be a promising option for patients in need of diaphragmatic reconstruction.