Abstract:
Background Bullous keratopathy is a major cause of corneal blindness, and anterior chamber injection of functional endothelial cells to substitute the diseased endothelium can effectively address the lack of donor source.
Objective To construct a modified model of rabbit bullous keratopathy with a preserved Descemet's membrane and evaluate the therapeutic effect of injecting rabbit primary corneal endothelial cell to the anterior chamber.
Methods Eighteen New Zealand rabbits weighing 2.5-3.0 kg were randomly divided into Con group and CEC group. A rabbit modified bullous keratopathy model was prepared using a self-designed silicone needle, 100 μL of cell-free DMEM + Y27632 was injected into the anterior chamber of the Con group, and 100 μL of rabbit primary corneal endothelial cells (containing 106 cells) + Y27632 were injected into the anterior chamber of the CEC group. The degree of corneal clarity and corneal thickness of rabbits after injection were evaluated by slit-lamp examination and anterior segment OCT at 1 d, 4 d, 7 d, and 14 d respectively. Corneal tissue was stained with alizarin red and hematoxylin-eosin at 14d to observe corneal pathological changes.
Results The corneal thickness showed no statistical difference between the CEC group and Con group at 1 d, 4 d, and 14 d, while at 7 d, the difference was significant (P ＜ 0.05) as the CEC group achieved clinical cure (corneal thickness ＜ 500 μm) in advance. The pathological results demonstrated that the CEC group had more intact corneal structure, with more tightly arranged stromal fibers, and regular morphology of hexagonal endothelial cells at 14 d.
Conclusion The modified bullous keratopathy model preparation method is simple and reliable, and the injection treatment of rabbit primary corneal endothelial cells into the anterior chamber is effective, which provides preclinical research results as well as a suitable animal model for the transformation and application of cell injection therapy.
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