Bartok Melinda, Tandon Rashmi, Alfaro-Espinoza Gabriela, Ullrich Matthias S, Gabel Detlef
Molecular Life Science Research Center, Jacobs University Bremen, Campus Ring 1, 28759, Bremen, Germany.
EXCLI J. 2015 Jan 21;14:123-32. doi: 10.17179/excli2014-556. eCollection 2015.
The irritative effects of preservatives found in ophthalmologic solution, or of antiseptics used for skin disinfection is a consistent problem for the patients. The reduction of the toxic effects of these compounds is desired. Brilliant Blue G (BBG) has shown to meet the expected effect in presence of benzalkonium chloride (BAK), a well known preservative in ophthalmic solutions, and octenidine dihydrochloride (Oct), used as antiseptic in skin and wound disinfection. BBG shows a significant protective effect on human corneal epithelial (HCE) cells against BAK and Oct toxicity, increasing the cell survival up to 51 % at the highest BAK or Oct concentration tested, which is 0.01 %, both at 30 min incubation. Although BBG is described as a P2x7 receptor antagonist, other selective P2x7 receptor antagonists, OxATP (adenosine 5'-triphosphate-2',3'-dialdehyde) and DPPH (N'-(3,5-dichloropyridin-4-yl)-3-phenylpropanehydrazide), did not reduce the cytotoxicity of neither BAK nor Oct. Therefore we assume that the protective effect of BBG is not due to its action on the P2x7 receptor. Brilliant Blue R (BBR), a dye similar to BBG, was also tested for protective effect on BAK and Oct toxicity. In presence of BAK no significant protective effect was observed. Instead, with Oct a comparable protective effect was seen with that of BBG. To assure that the bacteriostatic effect is not affected by the combinations of BAK/BBG, Oct/BBG and Oct/BBR, bacterial growth inhibition was analyzed on different Gram-negative and Gram-positive bacteria. All combinations of BAK or Oct with BBG hinder growth of Gram-positive bacteria. The combinations of 0.001 % Oct and BBR above 0.025 % do not hinder the growth of B. subtilis. For Gram-negative bacteria, BBG and BBR reduce, but do not abolish, the antimicrobial effect of BAK nor of Oct. In conclusion, the addition of BBG at bacterial inhibitory concentrations is suggested in the ready-to-use ophthalmic preparations and antiseptic solutions.
眼科溶液中发现的防腐剂或用于皮肤消毒的防腐剂的刺激性作用，对患者来说一直是个问题。人们期望降低这些化合物的毒性作用。亮蓝G（BBG）已显示在苯扎氯铵（BAK，眼科溶液中一种知名的防腐剂）和盐酸奥替尼啶（Oct，用于皮肤和伤口消毒的防腐剂）存在的情况下能达到预期效果。BBG对人角膜上皮（HCE）细胞显示出显著的保护作用，可抵御BAK和Oct的毒性，在最高测试浓度（0.01%）的BAK或Oct作用下孵育30分钟时，能将细胞存活率提高至51%。尽管BBG被描述为P2X7受体拮抗剂，但其他选择性P2X7受体拮抗剂，氧化三磷酸腺苷（OxATP）和N'-(3,5-二氯吡啶-4-基)-3-苯基丙烷酰肼（DPPH），均未降低BAK和Oct的细胞毒性。因此，我们认为BBG的保护作用并非因其对P2X7受体的作用。亮蓝R（BBR），一种与BBG类似的染料，也针对其对BAK和Oct毒性的保护作用进行了测试。在BAK存在的情况下未观察到显著的保护作用。相反，在Oct存在时，观察到了与BBG相当的保护作用。为确保抑菌效果不受BAK/BBG、Oct/BBG和Oct/BBR组合的影响，对不同革兰氏阴性菌和革兰氏阳性菌进行了细菌生长抑制分析。BAK或Oct与BBG的所有组合均会阻碍革兰氏阳性菌的生长。0.001%的Oct与高于0.025%的BBR组合不会阻碍枯草芽孢杆菌的生长。对于革兰氏阴性菌，BBG和BBR会降低但不会消除BAK和Oct的抗菌效果。总之。建议在即用型眼科制剂和防腐溶液中添加抑菌浓度的BBG。
