Quantitative microcomputed tomography analysis of collateral vessel development after ischemic injury
Quantitative microcomputed tomography analysis of collateral vessel development after ischemic injury
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DOI：
10.1152/ajpheart.00928.2003
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发表时间：
2004-07-01
影响因子：
4.8
通讯作者：
Guldberg, RE
中科院分区：
文献类型：
--
作者：
Duvall, CL;Taylor, WR;Guldberg, RE
转基因小鼠模型越来越多地用于研究特定生长因子或基质蛋白的功能，以设计控制血管生长的治疗策略。然而，用于评估这些模型中血管生成和动脉生成的可用方法受到动物大小、用户主观性、可视化三维血管网络的能力或采用强有力的定量分析的能力的限制。在这项研究中，我们采用对比增强微计算机断层扫描成像来评估诱导小鼠后肢缺血后的侧枝发育。术后 0、3 和 14 天对对照肢体和手术肢体的形态参数血管体积、连通性、数量、厚度、厚度分布、分离和各向异性程度进行评估。结果表明，手术操作肢体的血管容量早在股动脉切除后 3 天就通过一系列高度连接、小口径、紧密间隔且各向同性定向的侧支血管的发育而重建。完成参数分析以评估计算的形态参数对图像二值化阈值和体素大小变化的敏感性。研究发现，以 36 微米体素尺寸拍摄的图像对于评估侧支血管形成是最佳的，而需要 8 至 16 微米体素尺寸来解析较小的血管结构。这项研究证明了微计算机断层扫描作为血管网络定量、三维分析的稳健方法的实用性。虽然这些最初的努力集中在小鼠后肢缺血模型上，但所开发的技术可应用于各种模型系统以研究血管生成和动脉生成的机制。
Transgenic mouse models are increasingly being used to investigate the functions of specific growth factors or matrix proteins to design therapeutic strategies for controlling blood vessel growth. However, the available methodologies for evaluating angiogenesis and arteriogenesis in these models are limited by animal size, user subjectivity, the power to visualize the three-dimensional vessel networks, or the capability to employ a vigorous quantitative analysis. In this study, we employed contrast-enhanced microcomputed tomography imaging to assess collateral development after induction of hindlimb ischemia in the mouse. The morphological parameters vessel volume, connectivity, number, thickness, thickness distribution, separation, and degree of anisotropy were evaluated in control and surgery limbs 0, 3, and 14 days postsurgery. Results indicate that the vascular volume of the surgically manipulated limb was reconstituted as early as 3 days after femoral artery excision through development of a series of highly connected, small caliber, closely spaced, and isotropically oriented collateral vessels. Parametric analyses were completed to assess the sensitivity of the calculated morphological parameters to variations in image binarization threshold and voxel size. Images taken at the 36-mum voxel size were found to be optimal for evaluating collateral vessel formation, whereas 8- to 16-mum voxel sizes were needed to resolve smaller vascular structures. This study demonstrates the utility of microcomputed tomography as a robust method for quantitative, three-dimensional analysis of blood vessel networks. Whereas these initial efforts focused on the mouse hindlimb ischemia model, the developed techniques may be applied to a variety of model systems to investigate mechanisms of angiogenesis and arteriogenesis.