Ayaki Masahiko, Iwasawa Atsuo, Yaguchi Shigeo, Koide Ryohei
Department of Ophthalmology, Saitama National Hospital, Suma, Wako city, Saitama, Japan.
J Oleo Sci. 2011;60(3):139-44. doi: 10.5650/jos.60.139.
The aim of this study was to evaluate the cytotoxicity of anti-allergic eye drops for human corneal endothelial cells (HCEC) and commercially available ocular surface cells. A primary HCEC culture was derived from human eye bank specimens. SIRC (rabbit corneal epithelium), BCE C/D-1b (bovine corneal epithelial cells), RC-1 (rabbit corneal epithelium), and Chang (human conjunctival cells) were obtained commercially. The WST-1 assay was used to measure HCEC viability, and the viability of other cells was measured using the MTT assay. Cells were treated with 7 commercially available anti-allergic eye drops for 48 h and cell viability was measured and calculated as a percentage of control. The degree of toxicity for each eye-drop solution was based on the cell viability score (CVS). HCECs treated with a 1000-fold dilution of the eye-drop solution had a viability score of 67% for Rizaben and ≥80% for the other solutions with Zepelin being the least toxic. Cell viability in response to eye-drop solutions preserved with benzalkonium chloride (BAK) was dependent on the concentration of the drug solution and exposure time. Treatment of ocular surface cells with a 20-fold dilution of the eye-drop solution resulted in the following order of cell viability as determined by their CVS: Zepelin > Ketas = Zaditen ≥ Tramelas PF = Patanol ≥ Rizaben ≥ Livostin. This order was similar to that observed for HCECs, and cell viability was found to be concentration-dependent. Based on the penetration of the drug into eye tissues, HCECs are only likely to be pharmaceutically damaging in rare cases. Epithelial cell viability depends primarily on the concentration of BAK rather than on the action of the active component in the eye-drop solution. CVS values were useful for comparison of toxicity.
本研究的目的是评估抗过敏眼药水对人角膜内皮细胞（HCEC）和市售眼表细胞的细胞毒性。原代HCEC培养物来源于人眼库标本。SIRC（兔角膜上皮细胞）、BCE C/D - 1b（牛角膜上皮细胞）、RC - 1（兔角膜上皮细胞）和Chang（人结膜细胞）购自市售渠道。采用WST - 1法检测HCEC活力，其他细胞活力采用MTT法检测。用7种市售抗过敏眼药水处理细胞48小时，然后测量细胞活力并计算为对照的百分比。每种眼药水溶液的毒性程度基于细胞活力评分（CVS）。用1000倍稀释的眼药水溶液处理的HCEC，Rizaben的活力评分为67%，其他溶液的活力评分≥80%，其中Zepelin毒性最小。用含苯扎氯铵（BAK）保存的眼药水溶液处理眼表细胞时，细胞活力取决于药物溶液浓度和暴露时间。用20倍稀释的眼药水溶液处理眼表细胞后，根据CVS确定的细胞活力顺序如下：Zepelin > Ketas = Zaditen ≥ Tramelas PF = Patanol ≥ Rizaben ≥ Livostin。该顺序与HCEC观察到的顺序相似，且发现细胞活力具有浓度依赖性。基于药物在眼组织中的渗透情况，HCEC仅在极少数情况下可能受到药物损伤。上皮细胞活力主要取决于BAK的浓度，而非眼药水溶液中活性成分的作用。CVS值有助于毒性比较。
