Iontophoresis Transcorneal Delivery Technique for Transepithelial Corneal Collagen Crosslinking With Riboflavin in a Rabbit Model
Iontophoresis Transcorneal Delivery Technique for Transepithelial Corneal Collagen Crosslinking With Riboflavin in a Rabbit Model
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DOI：
10.1167/iovs.13-12595
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发表时间：
2016-02-01
影响因子：
4.4
通讯作者：
Malecaze, Francois
中科院分区：
文献类型：
--
作者：
Cassagne, Myriam;Laurent, Camille;Malecaze, Francois
目的.我们比较了离子电渗核黄素递送技术用于经上皮角膜胶原交联（I-CXL）与传统CXL（C-CXL）。我们设计了三个实验组，使用152只新西兰兔研究核黄素应用的离子电渗法使用带电核黄素溶液（Ricrolin+）与1 mA电流5分钟。第一组是比较离子电渗或常规核黄素应用后角膜中通过HPLC测量的核黄素浓度。第二组是通过使用非线性双光子显微镜（TP）和二次谐波发生（SHG）分析I-CXL或C-CXL后立即和14天的自体荧光和基质胶原蛋白修饰。在第三组中，通过应力-应变测量和研究角膜对胶原酶消化的抵抗力来评价I-CXL和C-CXL后的物理改变。基于HPLC分析，我们发现离子电渗允许核黄素扩散，其中核黄素浓度比常规应用低2倍（分别为936.2 +/- 312.5和1708 +/- 908.3 ng/mL，P < 0.05）。角膜TP和SHG成像显示，ICXL和C-CXL导致前基质和中基质自体荧光和胶原蛋白堆积的增加相当。10%应变下的应力显示，I-CXL或C-CXL治疗的角膜硬度相似（分别为631.9 +/- 241.5和680.3 +/- 216.4 kPa，P = 0.908）。此外，我们观察到I-CXL和C-CXL后角膜胶原酶消化的阻力增加（分别为61.90% +/- 5.28%和72.21% +/- 4.32%的剩余表面，P = 0.154）。该实验研究表明，I-CXL是交联治疗中核黄素递送的有前途的替代方法，保留了上皮。
PURPOSE. We compared an iontophoresis riboflavin delivery technique for transepithelial corneal collagen crosslinking (I-CXL) with a conventional CXL (C-CXL).METHODS. We designed three experimental sets using 152 New Zealand rabbits to study riboflavin application by iontophoresis using charged riboflavin solution (Ricrolin+) with a 1-mA current for 5 minutes. The first set was to compare riboflavin concentration measured by HPLC in corneas after iontophoresis or conventional riboflavin application. The second set was to analyze autofluorescence and stromal collagen modification immediately and 14 days after I-CXL or C-CXL, by using nonlinear two-photon microscopy (TP) and second harmonic generation (SHG). In the third set, physical modifications after I-CXL and C-CXL were evaluated by stress-strain measurements and by studying corneal resistance against collagenase digestion.RESULTS. Based on HPLC analysis, we found that iontophoresis allowed riboflavin diffusion with 2-fold less riboflavin concentration than conventional application (936.2 +/- 312.5 and 1708 +/- 908.3 ng/mL, respectively, P < 0.05). Corneal TP and SHG imaging revealed that ICXL and C-CXL resulted in a comparable increased anterior and median stromal autofluorescence and collagen packing. The stress at 10% strain showed a similar stiffness of corneas treated by I-CXL or C-CXL (631.9 +/- 241.5 and 680.3 +/- 216.4 kPa, respectively, P = 0.908). Moreover, we observed an increased resistance against corneal collagenase digestion after I-CXL and C-CXL (61.90% +/- 5.28% and 72.21% +/- 4.32% of remaining surface, respectively, P = 0.154).CONCLUSIONS. This experimental study suggests that I-CXL is a promising alternative methodology for riboflavin delivery in crosslinking treatments, preserving the epithelium.