University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, Texas, 75390-9057, USA.
Adv Ther. 2012 Oct;29(10):874-88. doi: 10.1007/s12325-012-0057-1. Epub 2012 Oct 10.
This in-vitro study compared the toxicity of bimatoprost 0.01% containing benzalkonium chloride (BAK) 0.02% with other commercial BAK-free or BAK-containing prostaglandin analogs.
Six test solutions were evaluated: travoprost 0.004% with polyquaternium-1 0.001% (PQ), PQ, bimatoprost 0.01% with BAK 0.02%, latanoprost 0.005% with BAK 0.02%, tafluprost 0.0015% preservative free (PF), and BAK 0.02%. Phosphate-buffered saline (PBS) was the live control and 70% methanol was the dead control. Confluent human corneal epithelial cells were incubated with test solutions (diluted 1:5 or 1:10 with PBS) or control solutions for 10 or 25 min, after which cells were fluorescently labeled to distinguish live and dead cells. Data were expressed as a percentage of PBS live-cell fluorescence for automated readouts. Live and dead cells were manually counted for numeric analyses.
For 1:5 and 1:10 dilutions using automated readout, cells exposed to bimatoprost with BAK, latanoprost with BAK, and BAK alone demonstrated significant reductions in the live cell signal compared with PBS, travoprost with PQ, and PQ alone (all P < 0.001). They also demonstrated significantly greater toxicity than tafluprost PF for 1:5 dilutions (all P < 0.001) and 1:10 dilutions (P ≤ 0.02), except for 1:10-diluted bimatoprost with BAK (P = 0.41). For 1:5 dilutions using manual cell count, cells exposed to bimatoprost with BAK demonstrated significant reductions in the percentage of live cells compared with PBS (P = 0.02). For 1:10 dilutions using manual cell count, cells exposed to bimatoprost with BAK, latanoprost with BAK, and BAK alone demonstrated significantly greater toxicity than PBS, travoprost with PQ, PQ alone, and tafluprost PF (all P ≤ 0.03). No significant differences were observed among PBS, travoprost with PQ, and PQ alone under any test conditions (P ≤ 0.63).
This study demonstrated that BAKcontaining solutions, including bimatoprost 0.01% with BAK, were toxic to human corneal epithelial cells, whereas BAK-free solutions showed little to no evidence of toxicity.
本体外研究比较了含有苯扎氯铵（BAK）0.02%的比马前列素 0.01%与其他商业 BAK 无或 BAK 含前列腺素类似物的毒性。
评估了六种测试溶液：含有多聚季铵盐-1 的 0.004%曲伏前列素（PQ）、PQ、含有 BAK 的 0.01%比马前列素、含有 BAK 的 0.005%拉坦前列素、0.0015%无防腐剂的他氟前列素（PF）和 BAK 0.02%。磷酸盐缓冲盐水（PBS）为活细胞对照，70%甲醇为死细胞对照。用测试溶液（用 PBS 稀释 1:5 或 1:10）或对照溶液孵育汇合的人角膜上皮细胞 10 或 25 分钟，之后用荧光标记物区分活细胞和死细胞。数据表示为自动读数的 PBS 活细胞荧光的百分比。手动计数活细胞和死细胞进行数值分析。
对于使用自动读数的 1:5 和 1:10 稀释，与 PBS、含有 PQ 的曲伏前列素和 PQ 相比，暴露于含有 BAK 的比马前列素、含有 BAK 的拉坦前列素和 BAK 单独的细胞的活细胞信号明显减少（均 P < 0.001）。与 tafluprost PF 相比，它们在 1:5 稀释（均 P < 0.001）和 1:10 稀释（P ≤ 0.02）时表现出更高的毒性，除了 1:10 稀释的含有 BAK 的比马前列素（P = 0.41）。对于使用手动细胞计数的 1:5 稀释，与 PBS 相比，暴露于含有 BAK 的比马前列素的细胞中活细胞的百分比明显减少（P = 0.02）。对于使用手动细胞计数的 1:10 稀释，与 PBS、含有 PQ 的曲伏前列素和 PQ 相比，暴露于含有 BAK 的比马前列素、含有 BAK 的拉坦前列素和 BAK 单独的细胞表现出更高的毒性（均 P ≤ 0.03）。在任何测试条件下，PBS、含有 PQ 的曲伏前列素和 PQ 单独之间均未观察到显著差异（P ≤ 0.63）。
本研究表明，含有 BAK 的溶液，包括含有 BAK 的 0.01%比马前列素，对人角膜上皮细胞有毒性，而不含 BAK 的溶液则几乎没有毒性证据。
