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移动激光间接检眼镜:用于在小鼠模型中诱导脉络膜新生血管形成

Mobile Laser Indirect Ophthalmoscope: For the Induction of Choroidal Neovascularization in a Mouse Model.

作者信息

Weinberger Dov, Bor-Shavit Elite, Barliya Tilda, Dahbash Mor, Kinrot Opher, Gaton Dan D, Nisgav Yael, Livnat Tami

机构信息

a Laboratory of Eye Research , Felsenstein Medical Research Center , Beilinson Campus, ‬Petach Tikva , Israel.

b Department of Ophthalmology , Rabin Medical Center , Beilinson Campus, Petach Tikva , Israel.

出版信息

Curr Eye Res. 2017 Nov;42(11):1545-1551. doi: 10.1080/02713683.2017.1349154. Epub 2017 Sep 21.

Abstract

PURPOSE

This study aims to evaluate and standardize the reliability of a mobile laser indirect ophthalmoscope in the induction of choroidal neovascularization (CNV) in a mouse model.

MATERIALS & METHODS: A diode laser indirect ophthalmoscope was used to induce CNV in pigmented male C57BL/6J mice. Standardization of spot size and laser intensity was determined using different aspheric lenses with increasing laser intensities applied around the optic disc. Development of CNV was evaluated 1, 5, and 14 days post laser application using fluorescein angiography (FA), histology, and choroidal flat mounts stained for the endothelial marker CD31 and FITC-dextran. Correlation between the number of laser hits to the number and size of developed CNV lesions was determined using flat mount choroid staining. The ability of intravitreally injected anti-human and anti-mouse VEGF antibodies to inhibit CNV induced by the mobile laser was evaluated.

RESULTS

Laser parameters were standardized on 350 mW for 100 msec, using the 90 diopter lens to accomplish the highest incidence of Bruch's membrane rupture. CNV lesions' formation was validated on days 5 and 14 post laser injury, though FA showed leakage on as early as day 1. The number of laser hits was significantly correlated with the CNV area. CNV growth was successfully inhibited by both anti-human and mouse VEGF antibodies.

CONCLUSION

The mobile laser indirect ophthalmoscope can serve as a feasible and a reliable alternative method for the CNV induction in a mouse model.

摘要

目的

本研究旨在评估并标准化一种便携式激光间接检眼镜在诱导小鼠脉络膜新生血管(CNV)模型中的可靠性。

材料与方法

使用二极管激光间接检眼镜在雄性色素沉着C57BL/6J小鼠中诱导CNV。使用不同的非球面透镜,在视盘周围施加逐渐增加的激光强度,确定光斑大小和激光强度的标准化。在激光照射后1、5和14天,使用荧光素血管造影(FA)、组织学以及用内皮标志物CD31和异硫氰酸荧光素标记的葡聚糖染色的脉络膜平铺片评估CNV的发展。使用脉络膜平铺片染色确定激光照射次数与已形成的CNV病变数量和大小之间的相关性。评估玻璃体内注射抗人及抗小鼠VEGF抗体抑制便携式激光诱导的CNV的能力。

结果

使用90屈光度透镜,将激光参数标准化为350 mW持续100毫秒,以实现最高的 Bruch 膜破裂发生率。激光损伤后第5天和第14天验证了CNV病变的形成,尽管FA早在第1天就显示有渗漏。激光照射次数与CNV面积显著相关。抗人及抗小鼠VEGF抗体均成功抑制了CNV的生长。

结论

便携式激光间接检眼镜可作为在小鼠模型中诱导CNV的一种可行且可靠的替代方法。

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