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

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.

Support of acellular porcine corneal stroma for growth of corneal epithelium and stromal cell in vitro / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To determine whether acellular porcine cornea stroma (APCS) could support the growth of the rabbit corneal cells in vitro.METHODS: APCS was prepared. The rabbit's corneal epithelium and stromal cells were cultured and seeded on, APCS in vitro.The observation of phase contrast photograph and histological examination were performed.RESULTS: Histological examination showed the epithe- lium grew on the scaffold of APCS in 2-3 layers at 10th day. The stromal cells adhered to the surface of the scaffold after 24 hours and invaded into the interlaminar of the material at 5th day.CONCLUSION: These results indicate that APCS can support the growth and proliferation of the corneal epithelium and stromal cells in vitro.

Management of corneal alkali burn with transplantation of limbal epithelial cells and decellularized porcine cornea in a rabbit model / 第三军医大学学报

Objective To investigate the therapeutic effect of transplantation of limbal epithelial cells and decellularized porcine cornea construct in rabbits with corneal alkali burn.Methods Allogeneic rabbit limbal epithelial cells were cultured and identified.Porcine cornea scaffold was fabricated by decellularization.The LECs-DPC construct was cultured and transplanted to the alkali-burned corneal surface of rabbits(4month follow-up).Results Immunocytochemistry and RT-PCR analyses revealed the presence of ABCG2 positive limbal stem cells.A general view showed the corneas were clear and vessel-free postoperatively.HE and Masson Trichrome staining results indicated the epithelial stratification,fewer cellular infiltration,DPC' integration into host corneal tissues,and more regular collagen configuration.Transmission electron microscopy exhibited formation of desmosomes and microvilli.Conclusion Allogeneic limbal epithelial cells transplantation using DPC as a carrier can restore corneal clarity after alkali burn in a rabbit model.