UV cross-linking of donor corneas confers resistance to keratolysis.
UV cross-linking of donor corneas confers resistance to keratolysis.
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DOI：
10.1097/ico.0000000000000185
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发表时间：
2014-09
期刊：
影响因子：
2.8
通讯作者：
Ciolino JB
中科院分区：
文献类型：
--
作者：
Arafat SN;Robert MC;Shukla AN;Dohlman CH;Chodosh J;Ciolino JB
开发一种改良的离体角膜交联方法，增加基质对酶降解的抵抗力，作为波士顿角膜假体的载体。使用Barron®人工前房进行人角膜的体外交联。角膜去上皮，核黄素溶液（0.1%核黄素/20%葡聚糖）预处理，UVA光（λ=370nm，辐照度=3mW/cm2）照射不同时间。UVA和γ照射的联合效应也通过市售的γ照射角膜供体进行评估。然后将角膜与0.3%的胶原酶A溶液在37摄氏度下孵育。比较不同治疗的角膜溶解时间。去上皮角膜（无紫外线照射，无核黄素）在5.8±0.6小时内溶解。交联角膜对溶解的抵抗力增加，溶解时间为17.8±2.6小时（p < 0.0001）。随着UVA暴露时间的延长，角膜组织对胶原酶的抵抗力增加，在30分钟时达到平稳期。与仅交联前角膜相比，交联前角膜和后角膜并不会增加阻力（p < 0.05）。无论是否进一步交联（5.6±1.2小时）（6.1±0.6小时）（p=0.43）， γ辐照的角膜与去上皮的对照组一样容易溶解。去上皮的前角膜表面胶原交联30分钟，对胶原酶A体外角化具有最佳的抵抗力。
To develop a modified ex vivo corneal crosslinking method that increases stromal resistance to enzymatic degradation for use as a carrier for the Boston keratoprosthesis. Ex vivo crosslinking of human corneas was performed using Barron® artificial anterior chambers. The corneas were de-epithelialized, pre-treated with riboflavin solution (0.1% riboflavin/20% dextran) and irradiated with ultraviolet A (UVA) light (λ=370nm, irradiance=3mW/cm2) for various durations. The combined effect of UVA and gamma (γ) irradiation was also assessed using the commercially available γ-irradiated corneal donors. The corneas were then trephined and incubated at 37 degrees Celsius with 0.3% collagenase A solution. The time to dissolution of each cornea was compared across treatments. De-epithelialized corneas (no UV light, no riboflavin) dissolved in 5.8 ± 0.6 hours. Crosslinked corneas demonstrated increased resistance to dissolution, with a time to dissolution of 17.8 +/− 2.6 hours (p < 0.0001). The corneal tissues’ resistance to collagenase increased with longer UVA exposure, reaching a plateau at 30 minutes. Crosslinking both the anterior and posterior corneas did not provide added resistance when compared to crosslinking the anterior corneas only (p>0.05). γ-irradiated corneas dissolved as readily as de-epithelialized controls regardless of whether they were further crosslinked (5.6 ± 1.2 hours) or not (6.1 ± 0.6 hours) (p=0.43) Collagen crosslinking of the de-epithelialized anterior cornea surface for 30 minutes conferred optimal resistance to in vitro keratolysis by collagenase A.