Extracellular matrix derived hydrogel for corneal tissue engineering

Abstract Purpose Several different synthetic and natural hydrogels have been suggested for use in corneal tissue engineering however these hydrogels lack many of the matrix constituents present in the real cornea. To overcome this problem we developed a new hydrogel derived from decellularized porcine cornea extracellular matrix for use in corneal tissue engineering. Methods Porcine corneas were decellularized using a combination of detergents and nucleases. The corneas were then freeze dried and milled into a fine powder. The powder was dissolved using a pepsin digest solution which underwent gelation at 37°C by neutralizing the solutions pH. Keratocytes were cultured on the hydrogels over several weeks to determine the hydrogels biocompatibility and ability to regulate cell behavior. Immunohistochemical staining was performed on the cell seeded hydrogels to examine the influence the hydrogel had on the cells phenotype. Results Decellularization of the corneal matrix was confirmed using a DNA assay prior to matrix digestion. The hydrogels that were formed using this technique were highly transparent and able to maintain viable cells. Keratocytes seeded onto the hydrogels had fully infiltrated them after 2 weeks. Markers of keratocyte cell behavior were observed after 2 weeks in culture suggesting that the hydrogel supports the maintenance of a keratocyte phenotype Conclusion We have demonstrated a new hydrogel that can be used for engineering corneal tissue. The next step will be to test corneal epithelial with these hydrogels to determine their suitability for corneal transplantation.