Tissue engineering a human conjunctiva-scleral model for in vitro testing

Abstract

My objective was to tissue engineer a human conjunctiva-scleral equivalent with very basic inflammatory components, using a combinatorial approach. The complete model would comprise an innervated, vascularised stroma, overlaid by a stratified epithelium within a bio-synthetic matrix. Matrices fabricated from fibrin and either poly (N-isopropylacrylamide) or poly (N-isopropylacrylamide)-co-acrylic acid, supported differentiation of a human vascular endothelial cell line I immortalized into vessel-like structures. Human neutrophils and a granulocytic cell line, HL60 were able to migrate through these matrices and produce matrix metalloproteinases, in response to chemotactic stimuli. Innervation was introduced by embedding dorsal root ganglia as nerve sources within the matrices, while epithelial cells were seeded on top of the matrix. Contributions of this thesis include: (1) methodology for tissue engineering a conjunctiva-scleral tissue substitute, and (2) demonstrating basic functionality. This model may be further developed for use as an alternative to animals for in vitro toxicology testing.