目的：研究曲安奈德（triamcinolone acetonide）懸浮液及其防腐劑苯甲醇（benzyl alcoho）對眼部組織的細胞毒性。
方法：眼角膜內皮細胞和視網膜色素上皮細胞培養自紐西蘭白兔，人類視網膜色素上皮細胞培養自ARPE-19細胞株。培養之細胞暴露於兩種市售含防腐劑之曲安奈德懸浮液（康寧克通Kenacort-A和信可得Shincort）、排除溶劑後之曲安奈德顆粒、純溶劑、十分之一稀釋之曲安奈德懸浮液、十分之一稀釋之排除溶劑後之曲安奈德顆粒、十分之一稀釋之純溶劑、或苯甲醇溶液（0.0225, 0.225, 0.9, 3, 9亳克／亳升），以眼科手術用平衡鹽類溶液做為控制組。暴露時間，對眼角膜內皮細胞為一至三十分鐘，而對視網膜色素上皮細胞則為五至一百二十分鐘。細胞型態改變以台盼藍（trypan blue）原位染色觀察；死亡細胞比率以台盼藍染劑排除分析法定量，功能性細胞以粒線體去氫酶分析法測定；超顯微構造則以穿透式電子顯微鏡觀察；細胞死亡機制以吖啶橙（acridine orange）和溴化乙錠（ethidium bromide）染色、去氧核醣核酸斷裂法（DNA laddering）、生物素缺口末端標記法染色（TUNEL）、碘化丙錠（propidium iodide）／annexin V-FITC／赫司特（Hoechst）33258染色、碘化丙錠／annexin V-FITC流式細胞儀、硫胱氨酸蛋白脢（caspase）活性分析、硫胱氨酸蛋白脢和細胞凋亡抑制分析、粒線體膜電位和西方墨點法評估。
結果：康寧克通、信可得和苯甲醇會以劑量和時間相關效應傷害培養之細胞，但排除溶劑後之曲安奈德顆粒則否。臨床上玻璃體內注射曲安奈德之苯甲醇相關濃度為0.225亳克／亳升，會在兩小時內導致人類視網膜色素上皮細胞之超顯微傷害和細胞功能受損，但0.0225亳克／亳升則否。而市售曲安奈德懸浮液中之苯甲醇9亳克／亳升，則只要五分鐘，在各種分析方法都會對人類或兔子視網膜色素上皮細胞造成毒性；甚至於只要一分鐘，就可對兔子眼角膜內皮細胞造成嚴重溶解。細胞死亡之主要機制是細胞急性壞死，細胞凋亡也有參與，而早期超顯微變化則是細胞質中胞器腫脹。
結論：市售含防腐劑之曲安奈德懸浮液和苯甲醇會傷害眼角膜內皮細胞和視網膜色素上皮細胞，然而去除防腐劑或溶劑之曲安奈德顆粒則不會。我們以細胞培養研究曲安奈德和苯甲醇細胞毒性之實驗具有重要臨床含意。為避免傷及眼角膜內皮細胞和視網膜色素上皮細胞，我們建議曲安奈德懸浮液注射於前房或玻璃體內之前，先排除其溶劑，特別是針對黃斑部裂孔手術。更進一步，我們期許研發不含防腐劑之曲安奈德懸浮液，並具生理性滲透壓和酸鹼值，以供眼內使用。

Purpose: to investigate the cytotoxicities of triamcinolone acetonide (TA) suspensions and their preservative, benzyl alcohol (BA), on ocular tissues.
Methods: Corneal endothelial cells and retinal pigment epithelial (RPE) cells were primarily cultured from New Zealand white rabbits, and human RPE cells were cultured from an RPE cell line, ARPE-19. Cultured cells were exposed to two commercially available preserved TA suspensions (Kenacort-A® and Shincort®), their vehicle-removed TA particles, pure vehicles, 1/10 dilutions of commercial TA suspensions, 1/10 dilutiuons of TA particles, 1/10 dilutions of pure vehicles, or BA solutions (0.0225, 0.225, 0.9, 3 or 9 mg/mL); the balanced salt solution was served as the control. The periods of exposure were 1 to 30 minutes for corneal endothelial cells and 5 to 120 minutes for RPE cells. Morphological changes of cells were evaluated by the trypan blue in situ staining. The proportions of dead cells were quantitatively measured by the trypan blue exclusion assay, and those of functional cells were assessed by a mitochondrial dehydrogenase assay. Ultrastructural changes were observed by transmission electron microscopy. The mechanism of cytotoxicity was determined by the acridine orange/ethidium bromide staining, DNA laddering technique, TUNEL staining, propidium iodide/annexin V-FITC/Hoechst 33258 staining, propidium iodide /annexin V-FITC flow cytometry, caspase activity assay, caspase and apoptotic inhibition assay, mitochondrial transmembrane potential assay, and Western blots.
Results: Kenacort-A®, Shincort®, their vehicles and BA damaged the cultured cells in dose- and time-dependent manners, but their vehicle-removed TA particles did not. BA 0.225 mg/mL, the clinically relevant concentration in TA following intravitreal injection, caused ultrastructural damage and impaired human RPE cell function at 2 hours; but BA 0.0225 mg/mL did not. BA 9.0 mg/mL, the concentration in commercial TA suspensions, was toxic within 5 minutes on each assay for both human and rabbit RPE cells, and even resulted in extensive lysis of rabbit corneal endothelial cells within only 1 minute. The major mechanism of cell death was necrosis, and apoptosis also played a minor role. The early ultrastructural change was swelling of organelles in the cytoplasm.
Conclusion: Preserved commercial TA suspensions and BA damaged corneal endothelial and RPE cells, but vehicle/preservative-free TA particles alone did not. Our in vitro studies on TA/BA cytotoxicity have significant clinical implications. To avoid damaging the corneal endothelial or RPE layer, we suggest removing the vehicle from TA suspensions before they are administered into the anterior chamber or vitreous cavity, particularly during macular hole surgery. Furthermore, we urge commercial development of a preservative-free TA suspension with a physiological osmolality and pH for intraocular use.

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