Buma P, Groenenberg M, Rijken P F, van den Berg W B, Joosten L, Peters H
Department of Orthopaedics, Orthopaedic Research Laboratory, University Medical Center, Nijmegen, The Netherlands.
Anat Rec. 2001 Apr 1;262(4):420-8. doi: 10.1002/ar.1050.
Many joint and bone diseases are caused by, or associated with vascular changes. Particularly in rheumatoid arthritis, vascular sprouting of synovial vessels plays a major role in the generation of joint pathology. To assess the effects of pharmaceuticals that are designed to inhibit neovascularization, we developed a quantitative procedure to measure vascular changes in cross-sections of the mouse knee joint during arthritic inflammation. Arthritis was induced in the knee joint of C57Black6 mice by a single subpatellar injection of methylated BSA after previous immunization. Total vascularity was visualized with a specific monoclonal rat anti-mouse antibody (9F1). Functional vessels were detected with the fluorescent perfusion marker Hoechst 33342. The localization of Hoechst and the vascular marker 9F1 were analyzed in separate images with an automated digital image processing system. By combining the two images, total vascularity and the perfusion status of the vessels during arthritis could be established. The digital image system measures synovial area (SA), number of all blood vessels (NBV) and the number of perfused blood vessels (NpBV). From these parameters the percentage of perfused vessels (perfusion fraction; PF), the vessel density (VD = NBV/SA) and the density of perfused vessels (VDp = NpBV/SA) can be calculated. The measurements showed that the area of synovial tissue had increased during arthritis. Moreover, both the number of blood vessels (NBV) and the number of perfused vessels (NpBV) in the synovial area had increased significantly on Days 4 and 7 after arthritis induction. This procedure enabled quantitation of total vascularity and of functional blood vessels in cross-sections of synovial tissue. It is expected to be a powerful tool, not only to analyze the effects of anti-angiogenic therapies in animal models of arthritis, but could also be applicable to study vascular and perfusion changes in vascular related diseases of the skeleton.
许多关节和骨骼疾病是由血管变化引起的，或与之相关。特别是在类风湿性关节炎中，滑膜血管的血管新生在关节病理形成中起主要作用。为了评估旨在抑制新血管形成的药物的效果，我们开发了一种定量程序，用于测量关节炎炎症期间小鼠膝关节横截面中的血管变化。在先前免疫后，通过髌下单次注射甲基化牛血清白蛋白在C57黑6小鼠的膝关节中诱导关节炎。用特异性单克隆大鼠抗小鼠抗体（9F1）使总血管可视化。用荧光灌注标记物Hoechst 33342检测功能性血管。使用自动数字图像处理系统在单独的图像中分析Hoechst和血管标记物9F1的定位。通过组合这两个图像，可以确定关节炎期间的总血管度和血管的灌注状态。数字图像系统测量滑膜面积（SA）、所有血管数量（NBV）和灌注血管数量（NpBV）。根据这些参数，可以计算灌注血管的百分比（灌注分数；PF）、血管密度（VD = NBV/SA）和灌注血管密度（VDp = NpBV/SA）。测量结果表明，关节炎期间滑膜组织面积增加。此外，在关节炎诱导后的第4天和第7天，滑膜区域的血管数量（NBV）和灌注血管数量（NpBV）均显著增加。该程序能够对滑膜组织横截面中的总血管度和功能性血管进行定量。预计它不仅是分析抗血管生成疗法在关节炎动物模型中的效果的有力工具，还可用于研究骨骼血管相关疾病中的血管和灌注变化。
