Objectives:The conversion of tissue engineering into a routine clinical tool cannot be achieved without a deep understanding of the interaction between cells and scaffolds during the process of tissue formation in an artificial environment. Here, we have investigated the cultivation conditions and structural features of the biodegradable non-woven material in order to obtain a well-differentiated human airway epithelium. Materials and methods:The bilayered scaffold was fabricated by electrospinning technology. The efficiency of the scaffold has been evaluated using MTT cell proliferation assay, histology, immunofluorescence and electron microscopy. Results:With the use of a copolymer of chitosan-gelatin-poly-l-lactide, a bilayered non-woven scaffold was generated and characterized. The optimal structural parameters of both layers for cell proliferation and differentiation were determined. The basal airway epithelial cells differentiated into ciliary and goblet cells and formed pseudostratified epithelial layer on the surface of the scaffold. In addition, keratinocytes formed a skin equivalent when seeded on the same scaffold. A comparative analysis of growth and differentiation for both types of epithelium was performed. Conclusions:The structural parameters of nanofibres should be selected experimentally depending on polymer composition. The major challenges on the way to obtain the well-differentiated equivalent of respiratory epithelium on non-woven scaffold include the following: the balance between scaffold permeability and thickness, proper combination of synthetic and natural components, and culture conditions sufficient for co-culturing of airway epithelial cells and fibroblasts. For generation of skin equivalent, the lack of diffusion is not so critical as for pseudostratified airway epithelium.
A unit for electrospinning of polymer melts was created. Nonwovens with an average fi bre diameter of 0.5-20 m were obtained from a melt of both pure polyamide 6 and its blends with stearic acid and oleic acid additives in the amount of 2-10 wt. % were obtained. Addition of 10% SF decreases the average diameter of the fi bres obtained by 40 times due to a decrease in the viscosity of the melt by 60 times.Electrospinning is a process for manufacturing ultrathin fi bres from polymer solution or melt under the effect of electrostatic forces. Electrohydrodynamic spraying of liquids (discovered for the fi rst time in 1745 by G. M. Bose [1]), where a weakly conducting liquid fl owing out of a nozzle under high voltage is sprayed into very fi ne drops by repulsive forces that are gradually precipitated on the opposite electrode, is the precursor of electrospinning. Patents for production of fi bre materials by electrospinning from solution were issued to Morton in 1902 in the USA for the fi rst time [2], but a major jump in the development of the solution method occurred in 1938 when I. V. Petryanov-Sokolov and N. D. Rozenblyum, colleagues at the L. Ya. Karpov Moscow Scientifi c-Research Institute of Physical Chemistry, in an attempt to obtain nitrocellulose aerosol from solution in acetone by electrohydrodynamic spraying, unexpectedly ran up against the competing regime of electrospinning of fi bres. Knowing how to evaluate the potential of this discovery, they could implement it in industry relatively rapidly.Fibres were probably obtained from polymer melts by the electrospinning method for the fi rst time in 1981 and described by Larondo and Manley [1]. They manufactured fi bres from polyethylene "solution-melt" in paraffi n and, more importantly, from pure polypropylene melt. The distance to the takeup plate was 1-3 cm, which allowed using an electric fi eld potential of approximately 7 kV without breakthrough of air. The fi bre diameter was greater than 50 m. It was then noted that the size of the fi bres obtained can be controlled by regulating the melt temperature and applied voltage. In addition, it was found that the viscosity is the most important parameter in production of fi bres from polymer melts than from solutions.The electrospinning process has many features. First, the one-time character of conducting all stages, so that a ready-to-use product is obtained at the end. Second, the universality of the method, which provides a broad spectrum of materials. Third, the fl exibility of the method, which allows controlling the structure of the materials. This is why the interest in production of polymer fi bres by electrospinning has now increased sharply.Despite the fact that most of the studies and manufacturing processes were executed for electrospinning of polymer solutions, spinning from polymer melts has a number of advantages.Many polymers (poly(ethylene terephthalate), polyolefi ns, some polyamides) are only soluble in scarce and expensive solvents at high temperature. This limits the use of these pol...
We analyzed viability of mesenchymal stem cells seeded by static and dynamic methods to highly porous fibrous 3D poly-L-lactide scaffolds with similar physical and chemical properties, but different spatial organization modified with collagen. Standard collagen coating promoted protein adsorption on the scaffold surface and improved adhesive properties of 100 μ-thick scaffolds. Modification of 600-μ scaffolds with collagen under pressure increased proliferative activity of mesenchymal stem cells seeded under static and dynamic (delivery of 100,000 cells in 10 ml medium in a perfusion system at a rate of 1 ml/min) conditions by 47 and 648%, respectively (measured after 120-h culturing by MTT test). Dynamic conditions provide more uniform distribution of collagen on scaffold fibers and promote cell penetration into 3D poly-L-lactide scaffolds with thickness >600 μ.
Biocompatibility of film and fibrous scaffolds from polylactide-based polymers and the relationship between their architecture and the functional characteristics of mesenchymal stem cells were studied. Cell culturing on polylactide-based film and fibrous matrixes did not deteriorate cell morphology and their proliferation and differentiation capacities. The rate of cell proliferation and penetration in microporous 3D matrices with the same porosity parameters and pore size depended on their spatial organization. The above materials can be used as scaffolds for mesenchymal stem cells for creation of tissue engineering implants. The scaffold size and structure should be determined by the defects in the organs in which the regeneration processes have to be stimulated.
The aim of this article is to verify the applicability of impulse acoustic microscopy as a tool for evaluating scaffolds in tissue engineering. Decellularized rat diaphragms and nonwoven polymer scaffolds made from cellulose diacetate, chlorinated polyvinyl chloride, and polysulfone were used as model objects. Optical microscopy, scanning electron microscopy, and histological research were used as reference methods in order to realize feasibility of the acoustic microscopy method in a regenerative medicine field. Direct comparison of different methods was carried out. The high frequency ultrasound microscopy is defined as a useful non‐invasive tool for studying and investigating the bulk morphology and bulk structure of opaque scaffolds, which allows to diagnose and to assume heterogeneous regions and defects inside the specimens. POLYM. ENG. SCI., 57:709–715, 2017. © 2017 Society of Plastics Engineers
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