Collagen Cross-Linking Using Rose Bengal and Green Light to Increase Corneal Stiffness
Collagen Cross-Linking Using Rose Bengal and Green Light to Increase Corneal Stiffness
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DOI：
10.1167/iovs.12-11509
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发表时间：
2013-05-01
影响因子：
4.4
通讯作者：
Kochevar, Irene E.
中科院分区：
文献类型：
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作者：
Cherfan, Daniel;Verter, E. Eri;Kochevar, Irene E.
目的。光化学交联角膜胶原蛋白是一种不断发展的治疗圆锥角膜和其他扩张性疾病。本研究采用玫瑰红加绿光（RGX）对兔眼胶原交联进行评价，并探讨其临床应用的重要因素。将红玫瑰（RB, 0.1%）涂抹于去核兔眼深层角膜2分钟。在冷冻切片上用荧光显微镜测定RB在基质中的扩散距离。将rb染色的角膜暴露在绿光（532 nm）下3.3 ~ 9.9分钟（50-150 J/cm(2)）。记录了辐照过程中吸收光谱的变化。采用单轴张力法测定角膜刚度。用布里渊显微镜观察基质弹性模量的空间分布。处理后24小时，在h&e染色切片上计数存活的角化细胞。RB在角膜基质中穿透约100 μ m，吸收了90%的入射绿光。RGX （150 J/cm(2)）使基质刚度增加3.8倍。间质前部弹性模量增加约120 μ m。在2分钟的照射过程中，RB被部分光漂白，但再次应用RB阻挡了70%的光透射。光谱测量表明RGX通过氧依赖机制引发交联。RGX不降低角化细胞活力。RGX在快速治疗（总时间约为12分钟）中显著增加角膜硬度，对角膜形成细胞没有毒性，可用于厚度小于400 μ m的角膜硬化。因此，RGX可能是抑制圆锥角膜和其他扩张性疾病进展的一种有吸引力的方法。
PURPOSE. Photochemical cross-linking of corneal collagen is an evolving treatment for keratoconus and other ectatic disorders. We evaluated collagen cross-linking by rose bengal plus green light (RGX) in rabbit eyes and investigated factors important for clinical application.METHODS. Rose bengal (RB, 0.1%) was applied to deepithelialized corneas of enucleated rabbit eyes for 2 minutes. The diffusion distance of RB into the stroma was measured by fluorescence microscopy on frozen sections. RB-stained corneas were exposed to green (532-nm) light for 3.3 to 9.9 minutes (50-150 J/cm(2)). Changes in the absorption spectrum during the irradiation were recorded. Corneal stiffness was measured by uniaxial tensiometry. The spatial distribution of the stromal elastic modulus was assessed by Brillouin microscopy. Viable keratocytes were counted on H&E-stained sections 24 hours posttreatment.RESULTS. RB penetrated approximately 100 mu m into the corneal stroma and absorbed >90% of the incident green light. RGX (150 J/cm(2)) increased stromal stiffness by 3.8-fold. The elastic modulus increased in the anterior approximately 120 mu m of stroma. RB was partially photobleached during the 2-minute irradiation, but reapplication of RB blocked light transmission by >70%. Spectral measurements suggested that RGX initiated cross-linking by an oxygen-dependent mechanism. RGX did not decrease keratocyte viability.CONCLUSIONS. RGX significantly increases cornea stiffness in a rapid treatment (congruent to 12 minutes total time), does not cause toxicity to keratocytes and may be used to stiffen corneas thinner than 400 mu m. Thus, RGX may provide an attractive approach to inhibit progression of keratoconus and other ectatic disorders.