Recreating the Microenvironment of the Native Cornea for Tissue Engineering Applications

Abstract

A viable tissue-engineered corneal replacement needs to be transparent and mechanically resilient. One necessary element for achieving this level of functionality is a scaffolding material that minimizes backscattered light, supports cellular growth, and maintains the transparent cellular phenotype. We hypothesize that the best scaffolding material will mimic the microenvironment of the natural corneal extracellular matrix (ECM). This work describes a method for electrospinning collagen type I fibers that replicates the unique morphology and arrangement of collagen type I fibers in the native cornea. In the cornea the collagen type I fibers are approximately 30 nm in diameter and aligned within stacked lamellae. After comparing several methods, the optimal method for creating uniformly aligned fibers was achieved by electrospinning onto a dual plate device with a quartz glass substrate. The fibers were crosslinked in glutaraldehyde vapor for 3 days and then further crosslinked and sterilized with liquid glutaraldehyde. Rabbit corneal fibroblasts were cultured on the fiber constructs for 7 days. Qualitative analysis of the cell morphology and intracellular protein expression suggests that the electrospun fibers provide a viable scaffold material for engineering a corneal tissue replacement.