Chang Yi-Sheng, Wu Chao-Liang, Tseng Sung-Huei, Kuo Pao-Ying, Tseng Shih-Ya
Department of Ophthalmology, College of Medicine, National Cheng Kung University, Taiwan.
Invest Ophthalmol Vis Sci. 2007 Jun;48(6):2792-8. doi: 10.1167/iovs.06-1146.
To investigate the toxic effects of triamcinolone acetonide (TA) suspensions on human retinal pigment epithelial (RPE) cells.
Cultured human RPE cells were exposed for up to 2 hours to one of seven solutions: control (balanced salt solution, BSS; Alcon Laboratories, Ft. Worth TX), commercial TA suspension (cTA), cTA from which the vehicle (which contains the preservative benzyl alcohol) had been removed (vehicle-removed TA, -vTA), vehicle of the cTA (V), or a 1:10 dilution (in BSS; Alcon) of cTA, -vTA or V. Solution effects were evaluated by phase-contrast microscopy of cells stained in situ with trypan blue and in vitro by trypan blue exclusion assay. RPE cell function was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The mechanism of TA toxicity was studied by acridine orange-ethidium bromide staining and epifluorescence microscopy, and ultrastructural changes were examined by transmission electron microscopy (TEM).
The effects of vehicle-removed solutions (-vTA and 1:10 -vTA) were similar to those of the control solution. Exposure for 1 hour or longer to a vehicle-containing solution (cTA and V) resulted in similar and significant degrees of cell damage that were dose and time dependent. The major mechanism of cell death was necrosis, and the early ultrastructural change was swelling of organelles in the cytoplasm.
Preserved commercial TA suspensions damaged human RPE cells, but vehicle-free solutions did not. The authors suggest removing the vehicle as completely as possible from TA solutions before they are administered intravitreally. Furthermore, they recommend that a commercial formulation of preservative-free TA suspension be made available for intraocular use.
研究曲安奈德（TA）混悬液对人视网膜色素上皮（RPE）细胞的毒性作用。
将培养的人RPE细胞暴露于七种溶液之一长达2小时：对照（平衡盐溶液，BSS；爱尔康实验室，得克萨斯州沃思堡）、市售TA混悬液（cTA）、去除了溶媒（含有防腐剂苯甲醇）的cTA（去除溶媒的TA，-vTA）、cTA的溶媒（V），或cTA、-vTA或V的1:10稀释液（在BSS中；爱尔康）。通过用台盼蓝原位染色的细胞相差显微镜检查和体外台盼蓝排斥试验评估溶液的作用。通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四氮唑（MTT）试验评估RPE细胞功能。通过吖啶橙-溴化乙锭染色和落射荧光显微镜研究TA毒性的机制，并通过透射电子显微镜（TEM）检查超微结构变化。
去除溶媒的溶液（-vTA和1:10 -vTA）的作用与对照溶液相似。暴露于含溶媒的溶液（cTA和V）1小时或更长时间会导致相似且显著程度的细胞损伤，且损伤程度与剂量和时间相关。细胞死亡的主要机制是坏死，早期超微结构变化是细胞质中细胞器肿胀。
保存的市售TA混悬液会损伤人RPE细胞，但无溶媒的溶液不会。作者建议在玻璃体内给药前尽可能完全地从TA溶液中去除溶媒。此外，他们建议提供一种不含防腐剂的TA混悬液的市售制剂用于眼内使用。
