ABSTRACT
The mechanical and physical properties of the cornea originate from the microstructure and composition of its extracellular matrix. It is known that collagen fibrils, with a relatively uniform diameter, are organized in a pseudo-hexagonal array. It has been suggested that proteoglycans and the interaction of their glycosaminoglycan (GAG) side chains with themselves and collagen fibrils are important for collagen fibril organization inside the cornea. There are several diseases such as keratoconus in which the regular collagen fibrillar packing becomes distorted causing corneal optical and mechanical properties to be compromised. The primary purpose of the present work was to investigate the role of GAGs on the microstructure of corneal extracellular matrix before and after corneal crosslinking (CXL) treatment. For this purpose, keratan sulphates (KS) were removed from corneal samples using the keratanase enzyme and the CXL procedure was used to crosslink the specimens. The transmission electron microscopy was then used to characterize the diameter of collagen fibrils and their interfibrillar spacing. It was found that KS GAG depletion increased the collagen interfibrillar spacing while the CXL treatment significantly decreased the interfibrillar spacing. The enzyme and CXL treatments had an insignificant effect on the diameter of collagen fibrils. The underlying mechanisms responsible for these observations were discussed in terms of the assumption that GAG chains form duplexes that behave as tiny ropes holding collagen fibrils in place.