NGF plus DHA treatment after PRK in rabbits is associated with increased corneal nerve surface area, increased epithelial proliferation, and decreased rose bengal staining compared with NGF, DHA, or vehicle control alone. The combination of NGF plus DHA yields faster nerve recovery after PRK and may have therapeutic usefulness in the treatment of post-PRK dry eye and other neurotrophic keratopathies.
Purpose-To assess ultrastructural modifications in keratocytes and inflammatory cell response in rabbit corneas after riboflavin and ultraviolet A (UVA) exposure using immunoflurescence microscopy.Methods-Twenty adult New Zealand albino rabbits weighing 2.0 to 3.0 kg were used in this study. Two rabbits served as controls. The animals had their epithelia removed and were cross-linked with riboflavin 0.1% solution (10mgs riboflavin-5-phosphate in 10ml 20% dextran-T-500) applied every 3 minutes for 30 minutes, and exposed to UVA (360 nm, 3 mW/cm 2 ) for 30 minutes. Four rabbits were humanely euthanized at each time point of 1, 3 and 11 days and at 3 and 5 weeks after the procedure.Immunohistochemestry studies of thin sections of each cornea were performed using TUNEL staining, Alpha smooth muscle actin (α-SMA), CD-3, myeloperoxidase (MPO) antibodies and DAPI counterstaining. In another experiment six additional rabbits were treated as above, and after 10 days of cross-linking, 5 μl of lipopolysaccharide (LPS) endotoxin (1μg/ml) was injected in the mid stroma.Results-Cross-linked corneas showed early stromal edema. By 5 weeks, complete resolution of the edema and a pronounced highly organized anterior 200 μm fluorescent zone was observed. TUNEL staining showed keratocyte death by both necrosis and apoptosis between day 1 and 3 after cross-linking. At day 1 the limbal area close to the cross linking zone showed some inflammatory cells as well as α-SMA positive cells, indicative of the presence of myofibroblasts. By day 3 some myofibroblasts had migrated to the area beneath the cross linked stroma. Between day 3 and 5 weeks there was an increase in α-SMA staining in the area surrounding the cross linked stroma. The area of cross linking remained acellular up to 5 weeks.
Conclusions-Collagen
At higher attempted corrections, LASEK-treated eyes showed less keratocyte apoptosis, myofibroblast transformation, and up-regulation in the synthesis of chondroitin sulfate than PRK-treated eyes. These differences may account for better visual acuities and less stromal haze in higher attempted corrections in LASEK-treated eyes.
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