Department of Ophthalmology, National Defense Medical College, Saitama, Japan.
Invest Ophthalmol Vis Sci. 2010 Aug;51(8):4337-45. doi: 10.1167/iovs.09-4707. Epub 2010 Mar 24.
To investigate the phototoxicity of persistent indocyanine green (ICG) under continuous visible light illumination and to determine whether blocking peak absorbance wavelengths of ICG is cytoprotective.
Cultured quail Müller cells were exposed to 0 to 5 mg/mL ICG for 30 seconds or 10 minutes and then were cultured in a colorless medium for 24 hours with or without continuous fluorescent lamp illumination. Cells exposed to 5 mg/mL ICG for 10 minutes were cultured under illumination filtered through a dichroic mirror that blocks red to near-infrared, green, or blue wavelengths. After microscopic observation, cell viability and cell death were evaluated.
ICG exposure followed by illuminated culture induced severe morphologic changes in cells, significant reductions in cell viability, and increases in cell death from apoptosis compared with exposure to ICG or illumination alone or with no exposure. Although ICG exposure at higher concentrations caused cell damage in a dose- and time-dependent manner, an increase in cell viability was noted for cells exposed to lower ICG concentrations. Blocking red to near-infrared wavelengths prevented the decrease in cell viability and the increase in cell death in the culture exposed to ICG followed by illuminated culture.
Continuous fluorescent lamp illumination enhanced the cytotoxicity of persistent ICG on Müller cells in a dose- and exposure time-dependent manner. Blocking peak absorbance wavelengths of ICG prevented photodynamic cytotoxicity of persistent ICG under continuous visible light illumination in vitro. This culture system could be used to study the mechanisms of prevention of unfavorable outcomes in ICG-assisted surgery.
研究持续可见光照下持久性吲哚菁绿(ICG)的光毒性,并确定是否能通过阻断 ICG 的峰值吸收波长来起到细胞保护作用。
将鹌鹑 Muller 细胞在 0 至 5mg/mL 的 ICG 中暴露 30 秒或 10 分钟,然后在无色培养基中培养 24 小时,同时进行或不进行连续荧光灯照射。将暴露于 5mg/mL ICG 10 分钟的细胞在经过二向色镜过滤的光照下培养,该二向色镜可以阻断红光到近红外光、绿光或蓝光。通过显微镜观察,评估细胞活力和细胞死亡情况。
ICG 暴露后进行光照培养会导致细胞形态发生严重变化,与单独暴露于 ICG 或光照或不暴露相比,细胞活力显著降低,细胞凋亡导致的细胞死亡增加。尽管较高浓度的 ICG 暴露会导致细胞损伤呈剂量和时间依赖性,但暴露于较低浓度的 ICG 会导致细胞活力增加。阻断红光到近红外光的波长可以防止在 ICG 暴露后进行光照培养时细胞活力的降低和细胞死亡的增加。
连续荧光灯照射会增强 Müller 细胞对持久性 ICG 的细胞毒性,呈剂量和暴露时间依赖性。阻断 ICG 的峰值吸收波长可以防止体外持续可见光照射下持久性 ICG 的光动力细胞毒性。这种培养系统可用于研究 ICG 辅助手术中不良结果的预防机制。