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多功能护理液对角膜上皮细胞形态及细胞因子产生的影响。

Effect of multipurpose solutions on cell morphology and cytokine production by corneal epithelial cells.

作者信息

Cole Nerida, Garthwaite Linda, Chen Renxun, Willcox Mark D P

机构信息

Brien Holden Vision Institute, The University of New South Wales, Sydney, New South Wales, Australia.

出版信息

Optom Vis Sci. 2012 Oct;89(10):1460-7. doi: 10.1097/OPX.0b013e318269c7b7.

Abstract

PURPOSE

We sought to determine whether human corneal limbal epithelial cells (HCLE) differ in their physiological response and production of inflammatory mediators during exposure to two multipurpose disinfecting solutions that differ in only four excipients.

METHODS

HCLE were exposed to dilutions (1%-20%) of OPTI-FREE Express and OPTI-FREE RepleniSH for 2, 6, or 18 h. Cell numbers were measured using CyQuant and metabolic activity assays. Morphology, viability, and apoptotic changes were examined by confocal microscopy after staining with Calcein AM/propidium iodide or Hoechst 33,342/propidiun iodide/YO-PRO-1. Cytokine responses and arachidonic acid metabolites were examined by enzyme-linked immunosorbent assay.

RESULTS

OPTI-FREE Express showed greater reductions in corneal cell metabolic activity (up to 3-fold) than OPTI-FREE RepleniSH over 18 h. Cells exposed to OPTI-FREE Express were highly vacuolated, whereas those exposed to OPTI-FREE RepleniSH had morphology consistent with a presumed apoptotic response. OPTI-FREE Express elicited higher levels of interleukin (IL)-6 (maximum 4225 ± 300 pg/mL) and IL-8 (maximum 1094 ± 250 pg/mL) from cells than OPTI-FREE RepleniSH (maximum of 1717 ± 225 pg/mL and 930 ± 300 pg/mL, respectively) at all concentrations tested after 18 h. HCLE did not produce leukotriene B4 or prostaglandin E2 on stimulation, and IL-1β was produced in low levels, but its production was not different between multipurpose disinfecting solution types.

CONCLUSIONS

The findings reported here are the first to describe the pattern of cytokine production produced in corneal epithelial cells in response to contact lens solutions. Alteration of only four excipients in the formulations had such effects on corneal epithelia; however, these differences do not explain differences in the incidences of corneal infiltrative events between these solutions. This might indicate that changes in the rates of infiltrative events result from interactions of the solutions with the contact lens surface, or directly with the corneal immune system, or as the result of gram-negative contamination of lens cases.

摘要

目的

我们试图确定人角膜缘上皮细胞(HCLE)在接触两种仅四种辅料不同的多功能消毒溶液时,其生理反应和炎症介质产生是否存在差异。

方法

将HCLE暴露于OPTI-FREE Express和OPTI-FREE RepleniSH的稀释液(1%-20%)中2、6或18小时。使用CyQuant和代谢活性测定法测量细胞数量。在用钙黄绿素AM/碘化丙啶或Hoechst 33,342/碘化丙啶/YO-PRO-1染色后,通过共聚焦显微镜检查形态、活力和凋亡变化。通过酶联免疫吸附测定法检测细胞因子反应和花生四烯酸代谢产物。

结果

在18小时内,OPTI-FREE Express比OPTI-FREE RepleniSH使角膜细胞代谢活性降低得更多(高达3倍)。暴露于OPTI-FREE Express的细胞高度空泡化,而暴露于OPTI-FREE RepleniSH的细胞形态与推测的凋亡反应一致。在18小时后测试的所有浓度下,OPTI-FREE Express从细胞中诱导产生的白细胞介素(IL)-6(最高4225±300 pg/mL)和IL-8(最高1094±250 pg/mL)水平高于OPTI-FREE RepleniSH(分别最高为1717±225 pg/mL和930±300 pg/mL)。HCLE在刺激后不产生白三烯B4或前列腺素E2,且IL-1β产生水平较低,但在不同类型的多功能消毒溶液之间其产生无差异。

结论

此处报道的研究结果首次描述了角膜上皮细胞对隐形眼镜护理液产生的细胞因子产生模式。制剂中仅四种辅料的改变就对角膜上皮产生了这样的影响;然而,这些差异并不能解释这些溶液之间角膜浸润事件发生率的差异。这可能表明浸润事件发生率的变化是溶液与隐形眼镜表面相互作用、或直接与角膜免疫系统相互作用的结果,或者是镜片盒革兰氏阴性菌污染的结果。

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