Fadlallah Ali, Zhu Hong, Arafat Samer, Kochevar Irene, Melki Samir, Ciolino Joseph B
Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, Massachusetts, United States 2Boston Eye Group, Brookline, Massachusetts, United States 3Wellman Center for Photomedicine, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, United States.
Wellman Center for Photomedicine, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, United States 4Department of Ophthalmology, Shanghai First People's Hospital, Shanghai JiaoTong University, Shanghai, China.
Invest Ophthalmol Vis Sci. 2016 Dec 1;57(15):6610-6614. doi: 10.1167/iovs.15-18764.
The purpose of this study was to evaluate the resistance to degradation by collagenase A of corneas that have been crosslinked with Rose Bengal and green light (RGX).
The ex vivo crosslinking procedure was performed on enucleated rabbit corneas. Corneas were deepithelialized after applying 30% alcohol. Corneas were stained with Rose Bengal (RB, 0.1%) for 2 minutes and then exposed to green light (532 nm) at 0.25 W/cm2 for times to deliver doses of 50, 100, 150, or 200 J/cm2 (n = 5 per group). Five corneas were pretreated with riboflavin solution (0.1% riboflavin) for 15 minutes and irradiated with ultraviolet A (UVA) light (370 nm, 3 mW/cm2) for 30 minutes. Five corneas underwent only de-epithelialization and were otherwise untreated. Five corneas were stained with RB without light exposure. The central corneas of each group was removed with a 8.5-mm trephine and incubated at 37°C in 0.3% collagenase A solution. Time to dissolution of each cornea was compared across treatments.
Corneas treated with RGX were treated with light fluences of 50, 100, 150, and 200 J/cm2; these corneas dissolved completely at 8.3 ± 1.2, 11.1 ± 1.4, 12.4 ± 1.7, and 15.7 ± 1.8 hours, respectively. Corneas treated by riboflavin and UVA light dissolved at 15.7 ± 1.7 hours, and nontreated corneas dissolved at 6.1 ± 1.3 hours. Corneas treated with only RB (no green light) dissolved at 9.3 ± 1.7 hours. Compared with the untreated corneas, all of the RB groups and the riboflavin-UVA-treated group of corneas degraded statistically significantly slower than untreated corneas (P < 0.05).
Crosslinking with RGX increased corneal resistance to digestion by collagenase comparable to that produced by riboflavin and UVA treatment.
本研究的目的是评估经孟加拉玫瑰红和绿光交联(RGX)的角膜对胶原酶A降解的抵抗能力。
对摘除的兔角膜进行离体交联程序。应用30%酒精后对角膜进行上皮去除。角膜用0.1%的孟加拉玫瑰红(RB)染色2分钟,然后以0.25W/cm²的强度暴露于绿光(532nm)下,照射时间分别为能产生50、100、150或200J/cm²的剂量(每组n = 5)。五片角膜用核黄素溶液(0.1%核黄素)预处理15分钟,并用紫外线A(UVA)光(370nm,3mW/cm²)照射30分钟。五片角膜仅进行上皮去除,未作其他处理。五片角膜用RB染色但未暴露于光下。每组的中央角膜用8.5mm的环钻取下,在37°C下于0.3%胶原酶A溶液中孵育。比较各处理组角膜溶解的时间。
用50、100、150和200J/cm²的光通量处理的RGX角膜,这些角膜分别在8.3±1.2、11.1±1.4、12.4±1.7和15.7±1.8小时完全溶解。用核黄素和UVA光处理的角膜在15.7±1.7小时溶解,未处理的角膜在6.1±1.3小时溶解。仅用RB处理(无绿光)的角膜在9.3±1.7小时溶解。与未处理的角膜相比,所有RB组和核黄素 - UVA处理组的角膜降解速度在统计学上均显著慢于未处理的角膜(P < 0.05)。
与核黄素和UVA处理产生的效果相当,RGX交联增加了角膜对胶原酶消化的抵抗能力。