The Study of Characteristics of Acellular Porcine Cornea Using Freezing-Thawing-Centrifugation

ABSTRACT

PURPOSE: To develop a new decellularization technique of porcine cornea using freezing-thawing-centrifugation (FTC) and to examine the characteristics of acellular porcine cornea (APC) for xenograft material. METHODS: Two-hundred micrometer thickness porcine corneas were decellularized with DNase/RNase, followed by 3 freezing-thawing-centrifugations (FTC, group 1), lyophilized FTC-APC (group 2), and chemical enzyme treated APC (CE-APC, group 3). Histologic evaluation to examine cells and collagen matrix, comparison of transparency, and cultivation to determine the viability of stromal cells was performed in fresh porcine cornea and 3 experimental groups. RESULTS: Decellularization occurred successfully in all experimental groups. Decellularization was confirmed by H&E staining and cultivation. Transparency of group 1 was similar to the normal porcine cornea but transparency of group 2 and group 3 was decreased. Collagen fibers of CE-APC (group 3) were not as well arrayed as FTC-APC (group 2). CONCLUSIONS: Acellularity of porcine cornea was successfully achieved by the FTC method with preservation of the cornea stroma. Novel decellularized porcine cornea can be considered as xenogeneic material for corneal transplantation.