Purpose: Collagen cross-linking using combined riboflavin/UVA treatment has been shown to increase the biomechanical rigidity of the cornea and has been used successfully for the treatment of progressive keratoconus. From morphological and biochemical investigations, a different degree of cross-linking for the anterior and posterior stroma by the treatment is suggested. The present study was undertaken to better evaluate this effect by testing the thermomechanical behavior. Methods: Ten 10 × 5 mm corneal strips from porcine cadaver eyes enucleated within 5 h post mortem were cross-linked using the photosensitizer riboflavin and UVA irradiation (370 nm, irradiance = 3 mW/cm2) for 30 min and compared to ten untreated corneal strips and ten corneal strips cross-linked with 0.1% glutaraldehyde. The temperature in a water bath was raised from 60 to 95°C with temperature increments of 1°C per minute. The hydrothermal shrinkage of the corneal strips was measured in 2.5°C steps using a micrometer. In addition, six 10-mm whole corneal buttons were cross-linked with riboflavin/UVA and immersed into water at 70 or 75°C. Results: The maximal hydrothermal shrinkage for the untreated control specimens and the posterior portion of the riboflavin/UVA-treated corneas was at 70°C, for the anterior portion of the cornea cross-linked by riboflavin/UVA at 75°C and for glutaraldehyde-cross-linked cornea at 90°C. In the cross-linked corneal buttons, a typical mushroom-like shape was observed at 70°C and a cylinder shape at 75°C. Conclusions: The different degree of collagen cross-linking in the corneal stroma after riboflavin/UVA treatment is reflected by the differences in the maximal shrinkage temperature of the anterior and posterior portion. Therefore, in the corneas cross-linked with riboflavin/UVA a higher shrinkage temperature was observed for the anterior portion of the cornea (75°C) compared to the posterior stroma (70°C) due to the higher degree of cross-linking of the anterior stroma. The anterior localization of the cross-linking effect is advantageous for the endothelium and for the preservation of the anterior corneal curvature.

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