[Xenogenic corneal acellular matrix as carrier for reconstruction of biological cornea epithelium-scaffold-endothelium compound].

OBJECTIVE

To investigate the method and feasibility of constructing biological cornea by culturing corneal epithelial and endothelial cells on the scaffold of xenogenic corneal acellular matrix (XCAM) in vitro.

METHODS

Porcine cornea was prepared as XCAM by application of detergent 1% Triton X-100 and freeze-drying process. After the carrier has rehydrated, rabbit epithelial and endothelial cells were seeded on each side of XCAM. After 2 weeks of culture, the reconstructed tissue of epithelium-scaffold-endothelium compound was examined by histological studies by HE staining and scanning electron microscope (SEM). The epithelium was examined by immunohistochemical studies using antibodies to cytokeratin (CK3), and the endothelium was stained with trypan blue and alizarin red.

RESULTS

Reconstructed biological cornea was composed of epithelium, acellular stroma and endothelium. Four to five layers of stratified flat cells were formed on the surface of XCAM, which were stained positively by CK3. Continuous monolayer cells located on the endothelial side, which were alive and showed honeycomb-like shape via dual staining with trypan blue and alizarin red, cells arranged tightly. Under SEM, epithelial cells showed several layers with the morphology of flat and spindle cells alternatively, endothelial cells showed polygonal shape with microvillus over the surface.

CONCLUSIONS

The biological corneal tissue reconstructed in vitro possessed three layers: the epithelium, scaffold and the endothelium. XCAM provides ideal surface for corneal epithelial and endothelial cells' adhesion and proliferation, it is desired to be used as scaffold for reconstruction of cornea in vitro.