Santen Oy, Tampere, Finland.
Curr Eye Res. 2012 Feb;37(2):145-54. doi: 10.3109/02713683.2011.626909. Epub 2011 Nov 3.
To investigate the cytotoxicity of benzalkonium chloride (BAC)-containing ophthalmic solutions of prostaglandin analogs (latanoprost, travoprost, bimatoprost, and preservative-free (PF) tafluprost), BAC mixture (BACmix) and BAC homologs with different alkyl chain lengths using human corneal epithelial (HCE) and conjunctival epithelial (IOBA-NHC) cell cultures. The distribution of BAC homologs in rabbit ocular surface tissues in vivo was examined.
The cells were exposed for one hour to prostaglandin analogs, BACmix and three homologs. Cytotoxicity was assessed with the WST-1 and lactate dehydrogenase (LDH) assays for cellular viability and cell membrane integrity. BAC 0.02% solution was instilled on the rabbit eye daily for 14 days and the concentrations of BAC homologs in external ocular tissues were determined.
The order of decreasing cytotoxicity in the WST-1 test was latanoprost ≥ travoprost > bimatoprost ≥ PF tafluprost. IOBA-NHC cells were more sensitive than HCE cells. In HCE, only latanoprost diluted to 10% increased LDH leakage. In IOBA-NHC, LDH leakage was statistically significant with 3-10% travoprost and 10% latanoprost. The order of decreasing cytotoxicity of preservatives was C14 > C12 > BACmix > C16 in HCE and C12 > C14 > BACmix > C16 in IOBA-NHC. Following treatment with BAC 0.02% solution, the amounts of BAC-C12, -C14 and -C16 in rabbit cornea and conjunctiva, respectively were: 0.37 ± 0.08 and 2.64 ± 0.27 ng/mg; 0.42 ± 0.07 and 4.77 ± 0.43 ng/mg; 0.04 ± 0.01 and 0.54 ± 0.05 ng/mg.
The cytotoxic effects of latanoprost, travoprost, and bimatoprost were dependent on the BAC concentration in their formulations. BACmix was cytotoxic at the concentrations above those corresponding to 0.001% BAC in ophthalmic medications. PF tafluprost was the least toxic of the drugs tested. Within studied BAC homologs, those with longer alkyl chain and higher lipophility penetrated effectively into rabbit external ocular tissues.
研究含有苯扎氯铵（BAC）的前列腺素类似物（拉坦前列素、曲伏前列素、贝美前列素和无防腐剂（PF）他氟前列素）、BAC 混合物（BACmix）和不同烷基链长的 BAC 同系物的眼部溶液对人角膜上皮（HCE）和结膜上皮（IOBA-NHC）细胞的细胞毒性。体内研究了 BAC 同系物在兔眼表面组织中的分布。
将细胞暴露于前列腺素类似物、BACmix 和三种同系物中 1 小时。用 WST-1 和乳酸脱氢酶（LDH）测定法评估细胞活力和细胞膜完整性的细胞毒性。每天将 BAC0.02%溶液滴注到兔子眼中 14 天，并测定外部眼组织中 BAC 同系物的浓度。
WST-1 试验中细胞毒性降低的顺序为拉坦前列素≥曲伏前列素>贝美前列素≥PF 他氟前列素。IOBA-NHC 细胞比 HCE 细胞更敏感。在 HCE 中，只有稀释至 10%的拉坦前列素增加 LDH 漏出。在 IOBA-NHC 中，3%-10%的曲伏前列素和 10%的拉坦前列素的 LDH 漏出具有统计学意义。在 HCE 和 IOBA-NHC 中，防腐剂的细胞毒性降低顺序为 C14>C12>BACmix>C16。用 BAC0.02%溶液处理后，兔角膜和结膜中的 BAC-C12、-C14 和 -C16 含量分别为：0.37±0.08 和 2.64±0.27ng/mg；0.42±0.07 和 4.77±0.43ng/mg；0.04±0.01 和 0.54±0.05ng/mg。
拉坦前列素、曲伏前列素和贝美前列素的细胞毒性作用取决于其制剂中 BAC 的浓度。BACmix 在高于眼科药物中 0.001% BAC 对应的浓度时具有细胞毒性。PF 他氟前列素是测试药物中毒性最小的。在所研究的 BAC 同系物中，烷基链较长且脂溶性较高的同系物能有效地渗透到兔眼外部组织中。
