Abstract
Purpose: :
To determine the efficiency of chitosan (CS) coated etafilcon A contact lens, as a substrate and carrier for rabbit limbal epithelial allograft.
Methods: :
Chitosan was coated on contact lens surface via polydopamine crosslinking. HCET cells were seeded on the surface of contact lens to study the cell attachment rate. Rabbit limbal epithelial tissue explants were co-cultured with 3T3 feeder cells on the CS-coated contact lens. The cell attachment and cytokeratin expression profile of rabbit epithelium was determined by light microscope and immunostaining. Tissue equivalents were tracked by DII and delivered to the surgically injured host rabbit to reconstruct the ocular surface. Cornea epithelium recovery, grafted cell proliferation, migration and differentiation were determined by slit-lamp examination and immunohistological analysis.
Results: :
CS-coated contact lens supported HCET cell attachment. Rabbit limbal epithelial cell proliferate and differentiate on the CS-coated contact lens in a co-culture system. Epithelial cells are able to form multilayer on the CS coated lens with strong immunoreactivity to CK3, CK12 and connexin 43. Moreover, CS-coated contact lens support the rapid delivery of epithelium layers onto the rabbit cornea. Grafted cells are proliferating (P63+ Ki67+) and differentiated (CK12+ Connexin43 +).
Conclusions: :
We modified the biocompatible CS-coated bandage contact lens to support the attachment, proliferation, delivery, and differentiation of rabbit limbal epithelial cells. This technique may be useful for the efficient delivery of the human corneal tissue equivalent to treat the patients with severe ocular surface diseases.
Keywords: contact lens • cornea: epithelium • immunohistochemistry