Abstract
Purpose: :
To evaluate the survival of decellularized human corneal graft in a mouse model.
Methods: :
Human cornea is decellularized by treating it with sodium chloride solution, followed by DNAse and RNAse solution. To test the efficacy of decellularized human cornea in vivo, graft re-cellularized with corneal cells or without cells is transplanted into the C57/BL6 mouse eye. Central cornea of 1.5 mm diameter and 70 μm depth is ablated with excimer laser from mouse eye and a thin section of decellularized cornea is grafted onto it. Mice are examined every day or every other day during the course of the study and are graded for neovascularization, opacity, inflammation, and edema with slit lamp microscope. At one week time point, eyes are enucleated and sections are stained with Hematoxylin and Eosin staining to assess the integration of donor tissue with the host tissue, and the fate of donor cells.
Results: :
The preliminary study is conducted for one week and the data suggested that the transplanted decellularized human cornea survived in the mouse eye during this time. In case of decellularized human cornea without the cells, host cells infiltrated the graft. While, in re-cellularized cornea, both host and donor cells migrated into and out of the graft assessed by staining cells with cells’ specific marker. However, the graft over time became slightly opaque but the transparency of the graft is not significantly affected.
Conclusions: :
This study evaluated the survival of decellularized human corneal graft with or without the corneal cells in a mouse model. Transplanted decellularized cornea survived in vivo for a period of one week with little or no graft rejection. The study will be extended to evaluate the long-term survival of decellularized graft in vivo.
Keywords: transplantation • cornea: basic science • cornea: stroma and keratocytes