Abstract
The goal of this proof-of-concept study was the fabrication of porous silk fibroin (SF) microspheres which could be used as cell culture carriers under very mild processing conditions. The SF solution was differentiated into droplets which were induced by a syringe needle in the high-voltage electrostatic field. They were collected and frozen in liquid nitrogen and water in droplets formed ice crystals which sublimated during lyophilization and a great quantity of micropores shaped in SF microspheres. Finally, the microspheres were treated in ethanol so as to transfer the molecular conformation into β-sheet and then they were insoluble in water. SF particles were spherical in shape with diameters in the range of 208.4 μm to 727.3 μm, while the pore size on the surface altered from 0.3 μm to 10.7 μm. In vitro, the performances of SF microspheres were assessed by culturing L-929 fibroblasts cells. Cells were observed to be tightly adhered and fully extended; also a large number of connections were established between cells. After 5-day culture, it could be observed under a confocal laser scanning microscope that the porous microenvironment offered by SF particles accelerated proliferation of cells significantly. Furthermore, porous SF particles with smaller diameters (200 - 300 μm) might promote cell growth better. These new porous SF microspheres hold a great potential for cell culture carriers and issue engineering scaffolds.
Highlights
Amplification of seed cells is the basis of tissue engineering, while microcarrier technology has realized the extensive culture of animal cells
The goal of this proof-of-concept study was the fabrication of porous silk fibroin (SF) microspheres which could be used as cell culture carriers under very mild processing conditions
Four groups of (a), (b), (c), and (d) microspheres with various diameters were obtained by changing electrostatic voltage and solution flow rate (Table 1) in this study
Summary
Amplification of seed cells is the basis of tissue engineering, while microcarrier technology has realized the extensive culture of animal cells. The higher specific surface area of microspheres could provide adequate adherent place for cells so as to be conducive to cell adhesion and proliferation. The current research about microcarriers focuses on polysaccharides, including cellulose, chitosan, hyaluronic acid, alginate, dextran and starch, as well as on proteins such as collagen, gelatin, elastin, albumin and silk fibroin [1,2]. SF materials can support the attachment, proliferation, and differentiation of primary cells and cell lines [5,6,7], and is prepared as films [8], porous scaffolds [9], gels [10], and tubular scaffolds [11]. The impressive cytocompatibility of SF materials make silk a popular starting material for tissue engineering scaffolds used in skin, bone, blood vessel, ligament, and nerve tissue regeneration [12,13,14]
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