A Quantitative and Standardized Method for the Evaluation of Choroidal Neovascularization Using MICRON III Fluorescein Angiograms in Rats.
A Quantitative and Standardized Method for the Evaluation of Choroidal Neovascularization Using MICRON III Fluorescein Angiograms in Rats.
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使用 MICRON III 荧光素血管造影评估大鼠脉络膜新生血管的定量和标准化方法。
DOI：
10.1371/journal.pone.0128418
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Yang D
中科院分区：
文献类型：
--
作者：
Wigg JP;Zhang H;Yang D
脉络膜新生血管（CNV）的体内成像在临床和基础研究中越来越被认为是研究年龄相关性黄斑变性（AMD）的一种有价值的工具。可以说，最广泛使用的复制AMD的模型是通过破裂Bruch膜在啮齿动物中激光产生的CNV。迄今为止，由于缺乏适当的啮齿类动物眼底相机和非标准化的分析方法，通过体内成像技术进行CNV评估一直受到阻碍。本研究旨在建立一种简单、可量化的眼底荧光素血管造影（FFA）图像分析CNV病变的方法。32只褐挪威大鼠；使用啮齿类动物专用眼底相机（Micron III， Phoenix实验室）在3周内拍摄FFA图像，并与传统的离体CNV评估进行比较。比较了腹腔注射和静脉注射荧光素获得的FFA图像，发现荧光素对病变性质有很大影响。利用常用的软件包，通过手动勾画每个病变的最大边界并对视神经头进行归一化，评估FFA图像对CNV和绒毛膜视网膜烧伤病变区域的影响。计算背景上净荧光和区域校正病变强度的导出值。在激光照射后2周，抗vegf抗体处理大鼠的CNV病变在归一化病变面积（p<0.001）和荧光强度（p<0.001）上明显小于PBS处理的对照组。在激光治疗后2周和3周，抗vegf治疗动物的计算面积校正病变强度显著小于（p<0.001）。使用FFA获得的结果与分离素染色脉络膜平板支架的常规病变面积测量结果相关，并得到证实，抗vegf治疗大鼠在激光治疗后2周（p = 0.049）和3周（p<0.001）的病变明显变小。本文提出的利用Micron III系统和常用软件进行CNV体内FFA定量的方法，包括采集变量校正，建立了一种可靠的CNV检测和定量方法，可以进行纵向研究，是传统CNV定量方法的重要替代方案。
In-vivo imaging of choroidal neovascularization (CNV) has been increasingly recognized as a valuable tool in the investigation of age-related macular degeneration (AMD) in both clinical and basic research applications. Arguably the most widely utilised model replicating AMD is laser generated CNV by rupture of Bruch’s membrane in rodents. Heretofore CNV evaluation via in-vivo imaging techniques has been hamstrung by a lack of appropriate rodent fundus camera and a non-standardised analysis method. The aim of this study was to establish a simple, quantifiable method of fluorescein fundus angiogram (FFA) image analysis for CNV lesions. Laser was applied to 32 Brown Norway Rats; FFA images were taken using a rodent specific fundus camera (Micron III, Phoenix Laboratories) over 3 weeks and compared to conventional ex-vivo CNV assessment. FFA images acquired with fluorescein administered by intraperitoneal injection and intravenous injection were compared and shown to greatly influence lesion properties. Utilising commonly used software packages, FFA images were assessed for CNV and chorioretinal burns lesion area by manually outlining the maximum border of each lesion and normalising against the optic nerve head. Net fluorescence above background and derived value of area corrected lesion intensity were calculated. CNV lesions of rats treated with anti-VEGF antibody were significantly smaller in normalised lesion area (p<0.001) and fluorescent intensity (p<0.001) than the PBS treated control two weeks post laser. The calculated area corrected lesion intensity was significantly smaller (p<0.001) in anti-VEGF treated animals at 2 and 3 weeks post laser. The results obtained using FFA correlated with, and were confirmed by conventional lesion area measurements from isolectin stained choroidal flatmounts, where lesions of anti-VEGF treated rats were significantly smaller at 2 weeks (p = 0.049) and 3 weeks (p<0.001) post laser. The presented method of in-vivo FFA quantification of CNV, including acquisition variable corrections, using the Micron III system and common use software establishes a reliable method for detecting and quantifying CNV enabling longitudinal studies and represents an important alternative to conventional CNV quantification methods.
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影响因子：
3.9
作者：
Schalenbourg, A.;Zografos, L.
通讯作者：
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影响因子：
4.4
作者：
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