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隐形眼镜护理液对人角膜上皮细胞的细胞毒性及对代谢的影响

Cytotoxicity and effects on metabolism of contact lens care solutions on human corneal epithelium cells.

作者信息

Choy Camus K M, Cho Pauline, Boost Maureen V

机构信息

School of Optometry, The Hong Kong Polytechnic University, Hong Kong SAR, China.

出版信息

Clin Exp Optom. 2012 Mar;95(2):198-206. doi: 10.1111/j.1444-0938.2011.00687.x. Epub 2012 Jan 11.

Abstract

PURPOSE

The aim was to determine the cytotoxic effects of three multipurpose solutions (MPS) on human corneal epithelial cells (HCEC) and to assess the metabolic rates of recovering cells at different levels of cell membrane damage.

METHOD

The effects of one to 15 minutes exposure to multipurpose solutions containing polyquaternium-1 (MPS-A), polyaminopropyl biguanide (MPS-B) and polyhexanide (MPS-C) on HCEC were determined. Recovery rates at different levels of cell membrane damage were assessed after re-culture for two hours at 37°C. Cell viability and membrane integrity were assessed using Annexin V-FITC/7-AAD staining and flow cytometry. Effects of concentrations of 10 to 40 per cent multipurpose solutions on the metabolic rate of recovering HCEC were assessed using a Vybrant MTT cell proliferation assay kit.

RESULTS

The highest percentage of late necrotic cells resulted after exposure to MPS-A compared with other solutions and the control (p < 0.001). The percentages of early necrotic cells after 10 and 15 minutes of soaking in MPS-B were significantly higher than the control and other multipurpose solutions (p < 0.001). Although MPS-C exposure also resulted in statistically significant higher percentages of early necrotic cells than the control (p < 0.005), these differences were small. No recovery was noted when HCEC treated with multipurpose solutions were re-cultured, with numbers of dead cells in MPS-B-treated cultures increasing fourfold. The MTT assay showed significant dose-response decreases of 500 nm absorbance for all MPS-treated cells. In 40 per cent MPS-A-treated HCEC, lack of activity indicated the cells were non-viable.

CONCLUSIONS

Multipurpose solutions induced varying levels of irreversible tissue sensitivity reactions, with MPS-A showing the greatest effects. The solutions damaged cell integrity and reduced metabolic rates suggesting delayed healing ability. The formulations of multipurpose solutions need to balance antimicrobial effectiveness with low cytotoxicity, which might not be currently possible to achieve. In light of our results, we suggest that contact lens wearers should be advised to rinse the soaked lenses with saline before lens insertion.

摘要

目的

本研究旨在确定三种多功能护理液(MPS)对人角膜上皮细胞(HCEC)的细胞毒性作用,并评估不同细胞膜损伤程度下恢复细胞的代谢率。

方法

测定含聚季铵盐-1(MPS-A)、聚氨基丙基双胍(MPS-B)和聚己缩胍(MPS-C)的多功能护理液作用于HCEC 1至15分钟后的效果。在37°C下再培养两小时后,评估不同细胞膜损伤程度下的恢复率。使用膜联蛋白V-异硫氰酸荧光素/7-氨基放线菌素D染色和流式细胞术评估细胞活力和膜完整性。使用Vybrant MTT细胞增殖检测试剂盒评估10%至40%浓度的多功能护理液对恢复中的HCEC代谢率的影响。

结果

与其他护理液和对照组相比,暴露于MPS-A后晚期坏死细胞的百分比最高(p < 0.001)。在MPS-B中浸泡10分钟和15分钟后,早期坏死细胞的百分比显著高于对照组和其他多功能护理液(p < 0.001)。虽然暴露于MPS-C后早期坏死细胞的百分比也显著高于对照组(p < 0.005),但差异较小。用多功能护理液处理的HCEC再培养后未观察到恢复,MPS-B处理的培养物中死细胞数量增加了四倍。MTT检测显示,所有MPS处理的细胞在500 nm吸光度上均有显著的剂量反应性下降。在40% MPS-A处理的HCEC中,缺乏活性表明细胞无活力。

结论

多功能护理液可引起不同程度的不可逆组织敏感性反应,其中MPS-A的影响最大。这些护理液破坏了细胞完整性并降低了代谢率,提示愈合能力延迟。多功能护理液的配方需要在抗菌效果和低细胞毒性之间取得平衡,而目前可能无法实现这一点。根据我们的研究结果,我们建议应建议隐形眼镜佩戴者在插入镜片前用生理盐水冲洗浸泡过的镜片。

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