Study on biocompatibility of complexes of collagen-chitosan-sodium hyaluronate and cornea

In this study we investigated the biocompatibility of collagen-chitosan-sodium hyaluronate (Col-Chi-NaHA) complexes and cornea tissue, and the feasibility of Col-Chi-NaHA complexes as substrates for cultivating rabbit corneal cells. Different components of Col-Chi-NaHA complexes were prepared and tested. A circular complex film with a diameter of 6 mm was inserted into rabbit stomal pocket and traced for a period of 5 months. Clinical examination was made. Rabbit limbal corneal epithelial cells, corneal endothelial cells, and keratocytes were cultured primarily on complexes. Phase contrast microscope examination was made daily. Histological, immunohistochemical, and scanning electron microscopic examinations were carried out. The complexes of 20% collagen, 10% chitosan, and 0.5% sodium hyaluronate showed rather weak corneal edema and other responses. The degradation of materials was obvious after 5 months. Corneas were transparent and translucent. Cells seeded on Col-Chi-NaHA were allowed to proliferate and partly form confluent monolayer after 9 days in culture. Cultured cells were well attached to the complexes of 20% collagen, 10% chitosan, and 0.5% sodium hyaluronate, or 10% chitosan and 0.5% sodium hyaluronate. The results showed that Col-Chi-NaHA complexes had good biocompatibility with cornea. The complexes can degrade and be absorbed in cornea. Col-Chi-NaHA complex may be a suitable substrate for cultivating corneal cells and a feasible material as a scaffold of tissue-engineered cornea.