*Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, MA; and †Schepens Eye Research Institute, Harvard Medical School, Boston, MA.
Cornea. 2014 Sep;33(9):955-9. doi: 10.1097/ICO.0000000000000185.
The aim of this study was to develop a modified ex vivo corneal cross-linking method that increases stromal resistance to enzymatic degradation for use as a carrier for the Boston keratoprosthesis.
Ex vivo cross-linking of human corneas was performed using Barron artificial anterior chambers. The corneas were deepithelialized, pretreated with riboflavin solution (0.1% riboflavin/20% dextran), and irradiated with ultraviolet A (UV-A) light (λ = 370 nm, irradiance = 3 mW/cm) for various durations. The combined effect of UV-A and gamma (γ) irradiation was also assessed using the commercially available γ-irradiated corneal donors. The corneas were then trephined and incubated at 37°C with 0.3% collagenase A solution. The time to dissolution of each cornea was compared across treatments.
Deepithelialized corneas (no UV light, no riboflavin) dissolved in 5.8 ± 0.6 hours. Cross-linked 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 UV-A exposure, reaching a plateau at 30 minutes. Cross-linking both the anterior and posterior corneas did not provide added resistance when compared with cross-linking the anterior corneas only (P > 0.05). γ-irradiated corneas dissolved as readily as deepithelialized controls regardless of whether they were further cross-linked (5.6 ± 1.2 hours) or not (6.1 ± 0.6 hours) (P = 0.43).
Collagen cross-linking of the deepithelialized anterior corneal surface for 30 minutes conferred optimal resistance to in vitro keratolysis by collagenase A.
本研究旨在开发一种改良的体外角膜交联方法,以增加基质对酶降解的抵抗力,用作波士顿角膜假体的载体。
使用 Barron 人工前房对离体角膜进行交联。角膜去上皮化后,用核黄素溶液(0.1%核黄素/20%葡聚糖)预处理,用紫外线 A(UV-A)光(λ=370nm,辐照度=3mW/cm)照射不同时间。还评估了商业上可用的γ辐照角膜供体的 UV-A 和γ联合照射的效果。然后,用角膜环钻钻取角膜,并在 37°C 下用 0.3%胶原酶 A 溶液孵育。比较各处理组角膜的溶解时间。
未用 UV 光和核黄素处理的去上皮化角膜在 5.8±0.6 小时内溶解。交联的角膜显示出对溶解的抵抗力增加,溶解时间为 17.8±2.6 小时(P<0.0001)。角膜组织对胶原酶的抵抗力随 UV-A 暴露时间的延长而增加,30 分钟后达到平台期。与仅交联前角膜相比,交联前、后角膜均未提供额外的抵抗力(P>0.05)。γ 辐照的角膜与去上皮化的对照物一样容易溶解,无论它们是否进一步交联(5.6±1.2 小时)或不交联(6.1±0.6 小时)(P=0.43)。
交联 30 分钟可使去上皮化的前角膜表面的胶原交联,从而使胶原酶 A 体外溶角膜的抵抗力最佳。