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 mi cropores 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 per formances of SF microspheres were assessed by culturing L-929 fibroblasts cells. Cells were observed to be tightly ad hered 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 parti cles 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.
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