Fibroblast growth factor-2 protects endothelial cells from damage after corneal storage at 4 degrees C.
BACKGROUND
Since the introduction of cold corneoscleral segment storage prior to keratoplasty there have been continuous efforts to ameliorate the preservation media in order to better maintain the quality of the corneal epi- and endothelium. Recent studies have shown that basic fibroblast growth factor (FGF-2) preserves the viability of, for example, retinal ganglion cells and pigment epithelium cells. Therefore, we investigated the effect of different concentrations of FGF-2 added to a modified Optisol storage medium on endothelial damage after corneal storage at 4 degrees C.


METHODS
. Bovine corneas were stored at 4 degrees C for 14 days and for another 24 h at 34 degrees C. Various FGF-2 concentrations (4, 20 and 40 ng/ml) were added to the medium either at day (D) 1, D14, or both D1 and D14. Quantitative evaluation of corneal damage after 14+1 days of storage was conducted by means of the Janus green photometry assay. Histological and ultrastructural investigations of the preserved endothelium were also performed. Bovine cell culture experiments using the TUNEL assay aimed to elucidate the role of FGF-2 on prevention of endothelial apoptosis.


RESULTS
The mean endothelial damage in control corneas increased from 4.9 +/- 1.8% (fresh corneas) to 13.4 +/- 2.4% after 14+1 days of storage. FGF-2 at 20 ng/ml or 40 ng/ml added at any of the indicated time points significantly reduced the overall endothelial damage by 5.1-7.3%, corresponding to 38-54% less endothelial damage than in control corneas (P<0.001). Light- and electron microscopic investigations confirmed this protective effect of FGF-2 on corneal endothelial cells. The TUNEL assay revealed a true anti-apoptotic effect of FGF-2 on endothelial cells in culture.


CONCLUSIONS
Our study clearly demonstrates the effectiveness of FGF-2 to enhance cell survival of the corneal endothelium after storage at 4 degrees C. A clinical interest could be seen in the potential future application of FGF-2 as an adjuvant to corneal preservation media in order to better maintain endothelial viability during corneal storage.
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002417:Cattle; D015925:Cryopreservation; D019610:Cytoprotection; D004305:Dose-Response Relationship, Drug; D004728:Endothelium, Corneal / Q000187:drug effects* / Q000473:pathology* / Q000503:physiopathology / Q000648:ultrastructure; D016222:Fibroblast Growth Factor 2 / Q000008:administration & dosage / Q000494:pharmacology*; D006801:Humans; D066298:In Vitro Techniques; D008855:Microscopy, Electron, Scanning; D009994:Osmolar Concentration; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000494:pharmacology
D011994:Recombinant Proteins; D016222:Fibroblast Growth Factor 2