Evaluation of Acute Corneal Barrier Change Induced by Topically Applied Preservatives Using Corneal Transepithelial Electric Resistance In Vivo
Evaluation of Acute Corneal Barrier Change Induced by Topically Applied Preservatives Using Corneal Transepithelial Electric Resistance In Vivo
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DOI：
10.1097/ico.0b013e3181a3c3e6
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发表时间：
2010-01-01
期刊：
影响因子：
2.8
通讯作者：
Kitaoka, Takashi
中科院分区：
文献类型：
--
作者：
Kusano, Mao;Uematsu, Masafumi;Kitaoka, Takashi
目的：我们评估了角膜屏障功能的急性变化后，滴注防腐剂使用角膜跨上皮电阻（TER）在体内和体外细胞毒性tests.Methods：角膜TER的活兔使用伏安表和银/银氯化物电极测量。将角膜暴露于防腐剂苯扎氯铵（BAC; 0.001%、0.002%、0.005%、0.01%和0.02%）、0.04%对羟基苯甲酸酯、0.5%氯丁醇、0.005%二葡萄糖酸氯己定、2%硼酸和0.01%乙二胺四乙酸中，然后监测TER的变化60秒。将培养的正常兔角膜上皮细胞在体外暴露于相同的防腐剂60秒，并使用WST-1测定法评估细胞活力。滴入0.01%和0.02%BAC后，TER立即下降，并在10秒内显著低于对照组0.005%BAC组60 s内与对照组比较差异有显著性（P < 0.01）。TER随BAC浓度的增加而降低。0.005%、0.01%和0.02%BAC滴注60秒后细胞活力显著低于对照组（P < 0.0001）。其他防腐剂均未显著改变TER或细胞活力。结论：BAC滴注后立即破坏角膜上皮，细胞存活率与TER的降低呈显著正相关（r = 0.94，P < 0.0001）。角膜上皮细胞死亡被认为与屏障功能下降有关;因此，体内角膜TER测量可以评估添加到眼科药物中的防腐剂的急性毒性。
Purpose: We evaluated acute changes in corneal barrier function after instillation with preservatives using corneal transepithelial electric resistance (TER) in vivo and cytotoxicity tests in vitro.Methods: The corneal TER of live rabbits was measured using a volt-ohm meter and silver/silver chloride electrodes. The cornea was exposed to the preservatives benzalkonium chloride (BAC; 0.001%, 0.002%,0.005%, 0.01%, and 0.02%),0.04% paraben, 0.5% chlorobutanol, 0.005% chlorhexidine digluconate, 2% boric acid, and 0.01% ethylenediaminetetraacetic acid, and then changes in the TER were monitored for 60 seconds. Cultured normal rabbit corneal epithelial cells were exposed to the same preservatives for 60 seconds in vitro, and cell viability was evaluated using the WST-1 assay.Results: The TER instantly decreased and became significantly lower than the control within 10 seconds after instillation with 0.01% and 0.02% BAC (P < 0.01) and within 60 seconds after that with 0.005% BAC (P < 0.01). The TER decreased concomitantly with increasing BAC concentration. Cell viability after instillation with 0.005%, 0.01%, and 0.02% BAC for 60 seconds was significantly lower than that of the control (P < 0.0001). None of the other preservatives significantly altered the TER or cell viability. Decreases in the TER correlated with cell viability (r = 0.94, P < 0.0001).Conclusions: Instillation with BAC immediately disrupted the corneal epithelium. Corneal epithelial cell death is supposed to be associated with a decline in barrier function; thus, corneal TER measurement in vivo can assess the acute toxicity of preservatives added to ophthalmic drugs.