Intravital Imaging to Understand Spatiotemporal Regulation of Osteogenesis and Angiogenesis in Cranial Defect Repair and Regeneration.
Intravital Imaging to Understand Spatiotemporal Regulation of Osteogenesis and Angiogenesis in Cranial Defect Repair and Regeneration.
复制标题
DOI：
10.1007/978-1-4939-8697-2_17
复制
发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Zhang X
中科院分区：
文献类型：
--
作者：
Zhang X
血管生成在骨骼修复和再生中起着至关重要的作用。由于缺乏有效的方法来在活体动物中以高时空分辨率同时跟踪骨愈合和新生血管形成，阻碍了我们对修复部位成骨和血管生成之间复杂关系的理解。为了克服这一障碍，我们最近在小鼠中建立了颅骨缺损窗室模型，该模型能够使用多光子激光扫描显微镜（MPLSM）对骨缺损愈合进行高分辨率、四维成像分析。窗口缺陷模型可通过 microCT 和 MPLSM 在体内对缺陷进行成像，从而促进修复过程中成骨和血管生成的谱系追踪和纵向分析。开窗室模型进一步允许插入细胞植入物或骨移植材料，有助于对活体动物中生物材料和血管微环境之间的相互作用进行时空分析。在本章中，我们将描述建立用于长期成像的慢性颅骨缺损窗室模型的改进技术，以及用于定量分析骨缺损修复部位的成骨和血管生成的成像分析协议。
Angiogenesis plays a critical role in skeletal repair and regeneration. Our understanding of the intricate relationship between osteogenesis and angiogenesis at a repair site has been hindered by the lack of an effective approach that allows tracking of bone healing and neovascularization simultaneously at a high spatiotemporal resolution in living animals. To overcome this barrier, we have recently established a cranial bone defect window chamber model in mice that enables high resolution, four-dimensional imaging analyses of bone defect healing using multiphoton laser scanning microscopy (MPLSM). The windowed defect model confers imaging of the defect through both microCT and MPLSM in vivo, facilitating lineage tracing and longitudinal analyses of osteogenesis and angiogenesis in repair. The windowed chamber model further permits insertion of cellular implants or bone graft materials, aiding in spatiotemporal analyses of the interactions between biomaterials and vascular microenvironment in living animals. In this chapter, we will describe the improved technique for establishing the chronic cranial defect window chamber model for long-term imaging as well as the imaging analysis protocols for quantitative analyses of osteogenesis and angiogenesis at the site of bone defect repair.