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M. Derse, P.O. Denk; Storage of Posterior Corneal Lamellae in a Cornea Bank . Invest. Ophthalmol. Vis. Sci. 2003;44(13):4710.
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Purpose: To evaluate the survival of corneal endothelial cells in posterior corneal lamellar buttons after long-term storage in a cornea bank compared to native full corneas under the same storage conditions. Goal of the study was to evaluate the risk for endothelial cell loss for pre-cutted posterior corneal lamellae. Methods: Freshly enucleated un-boiled porcine eyes were separated in 2 groups of 10 each. Eyes were rinsed in Polyvidon-Jod-Solution (10%). In all 10 eyes of group A a microkeratome cut of 180 microns was made using a commercially available microkeratome (Hansatome, Bausch & Lomb Surgical, Fl). The anterior flap was fully removed using a No. 15 scalpel blade. A corneo-scleral button was excised using a 15 mm trephine and the button was placed on a BOEHNKE-Chamber (Bausch & Lomb Surgical, Fl) in sterile culture medium I (Biochrom, Germany) for 3 weeks under 5 % CO2 and 95 % humidity. Eyes of group B served as controls and were treated in the same fashion except for the microkeratome cut. Before final examination stromal swelling of the buttons was reversed using culture medium II, which contains dextran. 3 digital photographs of the endothelial cells of each sample were made before and after the storage using a phase contrast microscope. Additionally the endothelium was stained using trypan blue and alizarine red S for further evaluation. Cells were counted using standard imaging software (Adobe Photoshop LE). In addition light microscopy of histological sections of the corneas was performed. Results: Endothelial count before storage was 3290 ± 256 cells/mm² in group A and 3300±245/mm² in group B. After storage time of 23 days the endothelial cell count decreased by 70 and 140 per mm² in group A and B, respectively. Neither this decrease nor the difference between the 2 groups was statistically significant (student's t-test). The observed decrease trend was less when the stained cells were used for comparison. An epithelial multilayer was found at the excision site in all eyes of groupA. Conclusions: When posterior corneal lamellae are long-term stored under organ culture conditions there is the potential risk of epithelial growth on the anterior surface, but endothelial cell density decreases only slightly. These data have to be confirmed for human corneas.
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