Silk fibroin from the Bombyx mori caterpillar has been processed into many material forms, with potential applications in areas ranging from optoelectronics to tissue engineering. As a hydrogel, silk fibroin has been engineered as a substrate for the regeneration of soft tissues where hydration and mechanical compatibility are necessary. Current fabrication of silk fibroin hydrogels produces microstructured materials that lack transparency and limits the ability to fully exploit the hydrogel form. Transparency is the main characteristic of some human tissues (e.g., cornea) where silk fibroin in the film format has shown potential as scaffolding material, however, lacking the necessary hydration and successful attachment of cells without biochemical functionalization. Additionally, detection using light is an important method to translate information for instruction, sensing, and visualization of biological entities and light sensitive molecules. Here, we introduce a method for the fabrication of transparent silk hydrogels by driving the formation of nanostructures in the silk fibroin material. These nanostructures are formed by exposing silk solution (concentration below 15 mg/mL) to organic solvents that induce the amorphous to crystalline transition of the protein and indeed the sol-gel transition of the material. We have also explored a process to modulate the mechanical properties of silk fibroin hydrogel within the physiological range by controlling the amount of metal ions present in the protein structure. Nanostructured silk fibroin hydrogels are biocompatible and allow for attachment and proliferation of human dermal fibroblasts without any biochemical functionalization. In addition, seeding of human cornea epithelial cells (HCECs) on the hydrogel surface results in the formation of an epithelium, which does not alter the gels' transparency and shows biological properties that challenge the performances of HCECs seeded in collagen hydrogels, the current standard material for the engineering of corneal tissue.
Keywords: cornea; fibroin; hydrogel; silk; transparency.