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高氧通透性硬性角膜接触镜兔模型中的铜绿假单胞菌感染性角膜炎。

Pseudomonas aeruginosa infectious keratitis in a high oxygen transmissible rigid contact lens rabbit model.

机构信息

Department of Ophthalmology, University of Texas Southwestern Medical Center, Dallas, Texas, United States.

出版信息

Invest Ophthalmol Vis Sci. 2014 Aug 14;55(9):5890-9. doi: 10.1167/iovs.14-14235.

Abstract

PURPOSE

To establish a rabbit model of infectious Pseudomonas aeruginosa keratitis using ultrahigh oxygen transmissible rigid lenses and characterize the frequency and severity of infection when compared to a non-oxygen transmissible lens material.

METHODS

Rabbits were fit with rigid lenses composed of ultrahigh and non-oxygen transmissible materials. Prior to wear, lenses were inoculated with an invasive corneal isolate of P. aeruginosa stably conjugated to green fluorescent protein (GFP). Corneas were examined before and after lens wear using a modified Heidelberg Rostock Tomograph in vivo confocal microscope. Viable bacteria adherent to unworn and worn lenses were assessed by standard plate counts. The presence of P. aeruginosa-GFP and myeloperoxidase-labeled neutrophils in infected corneal tissue was evaluated using laser scanning confocal microscopy.

RESULTS

The frequency and severity of infectious keratitis was significantly greater with inoculated ultrahigh oxygen transmissible lenses. Infection severity was associated with increasing neutrophil infiltration and in severe cases, corneal melting. In vivo confocal microscopic analysis of control corneas following lens wear confirmed that hypoxic lens wear was associated with mechanical surface damage, whereas no ocular surface damage was evident in the high-oxygen lens group.

CONCLUSIONS

These data indicate that in the absence of adequate tear clearance, the presence of P. aeruginosa trapped under the lens overrides the protective effects of oxygen on surface epithelial cells. These findings also suggest that alternative pathophysiological mechanisms exist whereby changes under the lens in the absence of frank hypoxic damage result in P. aeruginosa infection in the otherwise healthy corneal epithelium.

摘要

目的

利用超高透氧硬性隐形眼镜建立传染性绿脓假单胞菌角膜炎兔模型,并与非透氧隐形眼镜材料进行比较,以确定感染的频率和严重程度。

方法

将硬性隐形眼镜适配于兔子,这些隐形眼镜由超高透氧和非透氧材料制成。在佩戴之前,将镜片用稳定地与绿色荧光蛋白(GFP)偶联的侵袭性角膜绿脓假单胞菌分离株进行接种。使用改良的海德堡罗特斯特克体内共聚焦显微镜,在佩戴隐形眼镜前后对角膜进行检查。通过标准平板计数评估未佩戴和佩戴的隐形眼镜上粘附的活菌。使用激光扫描共聚焦显微镜评估感染性角膜组织中 GFP 标记的绿脓假单胞菌和髓过氧化物酶标记的中性粒细胞的存在情况。

结果

感染性角膜炎的发生率和严重程度在接种超高透氧隐形眼镜时明显更高。感染的严重程度与中性粒细胞浸润的增加有关,在严重的情况下,还会导致角膜融解。在佩戴隐形眼镜后对正常角膜进行体内共聚焦显微镜分析,证实缺氧隐形眼镜佩戴与机械表面损伤有关,而在高氧隐形眼镜组中则没有明显的眼部表面损伤。

结论

这些数据表明,在没有足够的泪液清除的情况下,被困在隐形眼镜下的绿脓假单胞菌的存在会抵消氧气对表面上皮细胞的保护作用。这些发现还表明,在没有明显缺氧损伤的情况下,在隐形眼镜下存在替代的病理生理机制,这会导致健康角膜上皮的绿脓假单胞菌感染。

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