Study of Human Epithelial Corneal Cells Regeneration on Silk-Fibroin-PEG Membranes

Abstract

The aim of the present work is to analyze the regeneration capabilities, as well as metabolic behavior and cell characterization of corneal epithelial cells on a Silk fibroin with 5% PEG membrane, before and after suffering an injury. The cells were analyzed and tested for up to 48 hours to properly assess wound regeneration. Microphotographs of these in vitro samples were realized every 24 hours and immediately after wound induction, cells reached confluency on the biomaterial substrate, as well as a physiological substrate. For cell characterization, we used the following immunohistochemistry: Cytokeratin 12 (CK12), Focal adhesion Kinase (FAK), αSMA, Phalloidin, and DAPPI. For metabolic essays, the WTS-1 test was used. To properly see the morphology Scanning and Transmission, Electronic Microscope images were acquired and analyzed. The Silk Membrane with 5% PEG was able to be as efficient as a simulated physiological ambient, as far as wound healing capacity goes, and showed promising signs of its use as a biomaterial having the same time to close wounds as other collagen-based materials. When the metabolic activity was regarded, it was possible to see a similar rise on both substrates showing again a good indicator for its clinical use. Nevertheless, future studies are needed to correctly determine the viability of this biomaterial in vivo models, for its consequences and methods of manipulation may widely differ for possible clinical use. However, it is possible to conclude that the Silk membrane with 5% PEG has great potential for future use in corneal damage.