Chemical-free fabrication of silk fibroin microspheres with silk I structure

Silk fibroin (SF) microspheres show bright prospects for biomedical applications, such as microcarriers, drug delivery, tumor embolization agents, and microscaffolds. However, the chemistry-independent preparation of SF microspheres, which is critical to biomedical applications, has been challenging. In this study, the SF microspheres with silk I crystal type were generated by using electrostatic spraying and freezing-induced assembly. The SF solution was sprayed into liquid nitrogen to form frozen microspheres with tunable size. Annealing can crystallize frozen SF to form silk I crystal type, providing a green approach to harvest water-insoluble microspheres. The SF microspheres can retain a monolithic shape in water for up to 30 days, while having a 77 % degradation ratio in PBS in 14 days, showing high stability in water and rapid degradation under physiological conditions. The biomedical application prospects of the silk I microspheres were demonstrated by cell culture and small molecule drugs (doxorubicin). The microspheres can support the growth and expansion of mammalian cells, and provide a sustainable release for DOX with 10 days. This strategy offers a green approach that avoids the use of organic solvents and cross-linkers for designing SF microsphere biomaterials.