One-Step Dip-Coating-Fabricated Core-Shell Silk Fibroin Rice Paper Fibrous Scaffolds for 3D Tumor Spheroid Formation.

Bioscaffolds are important substrates for supporting three-dimensional (3D) cell cultures. Silk fibroin (SF) is an attractive biomaterial in tissue engineering because of its good biocompatibility and mechanical properties. Electrospinning is one of the most often used approaches to fabricate SF fibrous scaffolds; yet, this technique still faces many challenges, such as low yield, residual organic solvents, limited extensibility of fibers, and a lack of spatial control over pore size. To circumvent these limitations, a core-shell SF on rice paper (SF@RP) fibrous scaffold was fabricated using a mild one-step dip-coating method. The cellulose fiber matrix of RP is the physical basis of the 3D scaffold, whereas the SF coating on the cellulose fiber controls the adhesion/spreading of the cells. The results indicated that by tuning the secondary structure of SF on the surface of a SF@RP scaffold, the cell behavior on SF@RP could be tuned. Tumor spheroids can be formed on SF@RP scaffolds with a dominant random secondary structure, in contrast to cells adhering and spreading on SF@RP scaffolds with a higher ratio of β-sheet secondary structures. Direct culturing of breast cancer MDA-MB-231 and MCF-7, lung cancer A549, prostate cancer DU145, and liver cancer HepG2 cells could spontaneously lead to corresponding tumor spheroids on SF@RP. In addition, the physiological characteristics of HepG2 tumor spheroids were investigated, and the results showed that compared with HepG2 monolayer cells, CYP3A4, CYP1A1, and albumin gene expression levels in HepG2 cell spheres formed on SF@RP scaffolds were significantly higher. Moreover, these spheroids showed higher drug resistance. In summary, these SF@RP scaffolds prepared by the dip-coating method are biocompatible substrates for cell culture, especially for tumor cell spheroid formation.