Becker M D, Kruse F E, Joussen A M, Rohrschneider K, Nobiling R, Gebhard M M, Völcker H E
Department of Ophthalmology, University of Heidelberg, Germany.
Graefes Arch Clin Exp Ophthalmol. 1998 May;236(5):390-8. doi: 10.1007/s004170050095.
The purpose of our study establish an animal model to study the microcirculation in corneal neovascularization in the living animal atraumatically.
Corneal neovascularization was induced in New Zealand white rabbits by a standard micropocket assay utilizing pellets with 250 ng basic fibroblast growth factor. Anesthesia consisted of intramuscular injections of ketamine and xylazine. Intravital microscopy was performed without preparation of the cornea. Rhodamine 6G was used as fluorescent dye to stain leukocytes. Fluorescein-isothiocyanate-dextran served as plasma marker. Microcirculation analysis was done off-line by digital video imaging with special analysis software and included the following parameters: vessel diameters, blood velocity, and differentiation of leukocytes according to their interaction with endothelium into free-floating. rolling and sticking leukocytes.
Vessel diameters in venular trunk vessels showed diameters of 54.0 +/- 13.3 microns with 1.1 +/- 0.5 mm/s flow; 29.4 +/- 16.3% of all leukocytes were attached to the vascular wall. The number of sticking leukocytes was found to be 17.8 +/- 36.0 cells/mm endothelial surface. Values are given for arteriolar trunk and branch as well as venular branch vessels.
This method for in vivo microscopic observation and quantification of the vasculature of the ocular surface seems to be suitable for evaluation of microhemodynamic and leukocyte measurements in mature neovascular vessels. It allows atraumatic experiments without corneal preparation procedures which disturb the microcirculation. The results concerning microhemodynamics and adherence of leukocytes are in a range comparable to other microcirculation studies. This new model could provide insight into the pathophysiology of microcirculatory disorders of the anterior eye segment, e.g. during angiogenesis.
本研究的目的是建立一种动物模型，用于在活体动物中无创地研究角膜新生血管形成中的微循环。
采用含250 ng碱性成纤维细胞生长因子的微球，通过标准的微囊试验在新西兰白兔中诱导角膜新生血管形成。麻醉采用肌肉注射氯胺酮和赛拉嗪。在不制备角膜的情况下进行活体显微镜检查。罗丹明6G用作荧光染料对白细胞进行染色。异硫氰酸荧光素-葡聚糖用作血浆标记物。通过数字视频成像和特殊分析软件离线进行微循环分析，包括以下参数：血管直径、血流速度，以及根据白细胞与内皮细胞的相互作用将其分为自由漂浮、滚动和黏附白细胞。
小静脉主干血管的直径为54.0±13.3微米，血流速度为1.1±0.5毫米/秒；所有白细胞中有29.4±16.3%附着于血管壁。发现黏附白细胞的数量为17.8±36.0个细胞/毫米内皮表面。给出了小动脉主干和分支以及小静脉分支血管的值。
这种用于体内显微镜观察和定量眼表血管系统的方法似乎适用于评估成熟新生血管中的微观血流动力学和白细胞测量。它允许进行无创实验，无需进行干扰微循环的角膜制备程序。关于微观血流动力学和白细胞黏附的结果与其他微循环研究的结果范围相当。这个新模型可以深入了解眼前节微循环障碍的病理生理学，例如在血管生成过程中。
