Quantitative assessment of angiogenesis and tumor vessel architecture by computer-assisted digital image analysis: Effects of VEGF-toxin conjugate on tumor microvessel density
Quantitative assessment of angiogenesis and tumor vessel architecture by computer-assisted digital image analysis: Effects of VEGF-toxin conjugate on tumor microvessel density
复制标题
DOI：
10.1006/mvre.1999.2233
复制
发表时间：
2000-05-01
影响因子：
3.1
通讯作者：
Griffioen, AW
中科院分区：
文献类型：
--
作者：
Wild, R;Ramakrishnan, S;Griffioen, AW
肿瘤生长依赖于血管生成。因此，旨在破坏这一机制的策略受到了大量研究。几种血管生成测定用于直接比较抗血管生成化合物的功效。然而，新血管生长的客观评估一直难以实现。本研究的目的是测试和开发一种计算机辅助图像分析方法，该方法可以无偏地定量微血管密度。人肿瘤在无胸腺小鼠中生长，并且在用VEGF-毒素缀合物处理一周后进行肿瘤活检。制备冷冻肿瘤切片，并用PE缀合的抗CD-31抗体染色，并用荧光显微镜对血管成像。通过量化每个记录视野的PE阳性像素来分析血管密度。此外，通过自动二值化和二值化方案进一步处理图像以研究形态学差异。该程序允许计算机辅助估计重要的血管生成参数，如总血管数、长度和分支点。基于这些指数，容易检测到对照肿瘤和用VEGF-毒素缀合物处理的肿瘤之间的血管生成反应的差异（对于所有参数，P < 0.007）。更重要的是，计算机生成的测量值与手动微血管计数相关性良好，并且显示出显著更少的变化。我们的研究结果表明，计算机辅助图像分析是一种快速，客观，和替代方法的定量评估肿瘤血管生成和血管结构。(C)中国科学出版社.
Tumor growth is angiogenesis dependent. As a consequence, strategies aimed at disrupting this mechanism are heavily investigated. Several angiogenesis assays are used to directly compare the efficacy of anti-angiogenic compounds. However, objective assessment of new vascular growth has been difficult to achieve. The aim of this study was to test and develop a computer-assisted image analysis method that would give an unbiased quantification of the microvessel density. Human tumors were grown in athymic mice and tumor biopsies were taken after a weeklong treatment with VEGF-toxin conjugate. Frozen tumor sections were prepared and stained with PE-conjugated anti-CD-31 antibodies and vessels were imaged with a fluorescence microscope. Vessel density was analyzed by quantifying PE-positive pixels per recorded field. In addition, images were further processed to investigate morphological differences by an automated binarization and skeletonization protocol. This procedure allowed the computer-assisted estimation of important angiogenic parameters such as total vessel number, length, and branch points. Based on these indices, differences in the angiogenic response between control tumors and those treated with VEGF-toxin conjugate were readily detected (P < 0.007 for all parameters). More importantly, computer-generated measurements correlated well with manual microvessel counts and showed significantly less variation. Our results suggest that computer-assisted image analysis represents a rapid, objective, and alternative method for the quantitative assessment of tumor angiogenesis and vessel architecture. (C) Academic Press.