Analysis of osteo-immune crosstalk during bone healing by longitudinal intravital imaging

纵向活体成像分析骨愈合过程中的骨免疫串扰

• 批准号: 259547803
• 负责人: Professorin Dr. Anja Erika Hauser
• 金额: --
• 依托单位: Deutsches Rheuma-Forschungszentrum Berlin (DRFZ)
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2014
• 资助国家: 德国
• 起止时间: 2013-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/259547803?language=en
• 关键词: Analysis; osteo; immune; crosstalk; during

Fracture healing is initiated with an inflammatory phase, during which immune cells invade the damaged tissue area. It is followed by a phase of bone repair. Both processes involve highly dynamic processes on the cellular and subcellular level, such as immune cell migration and cellular interactions between immune and stromal compartments. Intravital microscopy is the method of choice to study these processes, however, analyzing the spatiotemporal aspects of bone regeneration longitudinally has not been possible until now. To overcome this challenge, we have developed a microendoscopic device (LIMB) in the first funding period, allowing to analyze osteo-immune interactions during bone healing in vivo by repeated imaging of the same tissue region in the bone marrow. In the second funding period, we aim to use LIMB to dissect the role of aging on stromal and immune compartments by analyzing their dynamics in different mouse models of fracture healing. The impact of immune cell-derived factors will be tested using conditional knockout mouse strains. Differentiation of immune and mesenchymal cells will be monitored via the analysis of cellular metabolism by microendoscopic fluorescence lifetime imaging. Angiogenesis, which is known to be spatially and mechanistically linked to bone formation, will be analyzed in different age cohorts using LIMB.

骨折愈合始于炎症期，在此期间免疫细胞侵入受损组织区域。随后是一个骨修复阶段。这两个过程都涉及细胞和亚细胞水平上的高度动态过程，如免疫细胞迁移和免疫细胞与间质室之间的细胞相互作用。活体显微镜是研究这些过程的首选方法，然而，直到现在，纵向分析骨再生的时空方面还不可能。为了克服这一挑战，我们在第一个资助期开发了一种显微内窥镜设备（LIMB），允许通过对骨髓中同一组织区域的重复成像来分析体内骨愈合过程中的骨免疫相互作用。在第二个资助期，我们的目标是通过分析不同小鼠骨折愈合模型中基质和免疫室的动力学，利用LIMB来解剖衰老对基质和免疫室的作用。免疫细胞衍生因子的影响将使用条件敲除小鼠品系进行测试。免疫细胞和间充质细胞的分化将通过显微内窥镜荧光寿命成像分析细胞代谢来监测。血管生成，已知在空间和机制上与骨形成有关，将在不同年龄的队列中使用肢体进行分析。