Optimization of silk films as substrate for functional corneal epithelium growth.

The corneal epithelium is the first cellular barrier to protect the cornea. Thus, functional tissue engineering of the corneal epithelium is a strategy for clinical transplantation. In this study, the optimization of silk films (SFs) as substrates for functional human corneal epithelium growth was investigated with primary human corneal epithelial cells on SFs, poly-D-lysine (PDL) coated SFs, arginine-glycine-aspartic acid (RGD) modified SFs and PDL blended SFs. PDL coated SFs significantly promoted cell adhesion at early phases in comparison to the other study groups, while PDL blended SF significantly promoted cell migration in a "wound healing" model. All film modifications promoted cell proliferation and viability, and a multi-layered epithelium was achieved in 4 weeks of culture. The epithelia formed were tightly apposed and maintained an intact barrier function against rose bengal dye penetration. The results suggested that a differentiated human corneal epithelium can be established with primary corneal epithelial cells on SFs in vitro, by optimizing SF composition with PDL.