Enzymatic evidence of the depth dependence of stiffening on riboflavin/UVA treated corneas

Enzymatic evidence of the depth dependence of stiffening on riboflavin/UVA treated corneas
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DOI:
10.1007/s00347-007-1587-9
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发表时间:
2008-02-01
期刊:
影响因子:
--
通讯作者:
Pillunat, L. E.
Pillunat, L. E.
中科院分区:
医学4区
文献类型:
--
作者:
Schilde, T.;Kohlhaas, M.;Pillunat, L. E.

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Purpose. It has been shown that the treatment of keratoconus with riboflavin/ultraviolet A (UVA) causes significant stiffening of the cornea due to cross-linking. The aim of this study was to evaluate how deep the mechanical stabilization after collagen cross-linking could be shown biochemically.Method. Ten out of 20 enucleated porcine eyes were treated with riboflavin as a photosensitizer and UVA (370 nm, 3 mW/cm(2), 30 min). The other 10 eyes served as controls. With a Microkeratom device, two flaps with a thickness of 200 mu m and a diameter of 8 mm were cut off from each eye and put in a collagenase solution (NaCl plus collagenase A, 1: 1). The surfaces of the flaps were measured digitally every day to characterize the dissolving behavior.Results. The resistance ( regarding corneal collagen against enzymatic digestion) of the treated superficial flaps was considerably higher (p=0.001) compared to those that were cut secondarily and to the control flaps. But even the flaps from deeper layers showed a significant increase in resistance (p=0.02) compared with the untreated flaps. The half-life of the surfaces of the treated superficial flaps was 220 h; of those cut secondarily, it was 80 h. Both untreated flaps had a half-life of 50 h.Conclusions. The biochemical study showed that the treatment of the cornea with riboflavin/UVA leads to significant collagen cross-linking not only in the anterior slice of 200 mu m but also in the following 200 mu m. This locally limited cross-linking effect may be explained by the absorption behavior for UVA of the riboflavin-treated cornea; 65% of UVA irradiation is absorbed in the first 200 mu m and only 25-30% in the next 200 mu m. Therefore, deeper-lying structures and especially the endothelium are not affected.