Ex Vivo Keratoconus Model for TDP Analysis

Fresh albino rabbit eyes were purchased from Pelfreez Biologicals (Rogers, AR, USA). The ectatic model was previous developed in our lab [21 (link)], briefly, to create the ex vivo keratoconus-like model (KC) corneas, the eyes were treated with 0.1 U/mL chondroitinase ABC (C36675U Sigma Aldrich, St. Louis, MO, USA) by immersing the anterior surface in enzyme solution for 2 h. Following enzyme treatment, the eye globes were washed with full medium several times. The KC corneas were then treated to the different TDPs by submersing the anterior segment in TDP-suspended medium containing 4% dextran-500 (Sigma Aldrich, St. Louis, MO, USA) for 18 h. The dextran was added to reduce extensive swelling of the ex vivo corneas. Control corneas without enzyme treatment were cultured in 4% dextran full medium without TDP microparticles.