Advanced Optical Imaging of 3-D Cell Dynamics in Engineered Skin

1033906 Boppart尽管在过去十年中取得了重要进展，但目前的组织工程皮肤等同物对于需要可行皮肤替代品的患者来说只不过是一种脆弱的短期敷料。目前的皮肤替代品的主要弱点是组成细胞是在与天然皮肤非常不同的条件下培养和应用的。据认为，这主要是由于我们有限的理解的作用，3-D支架地形和机械刺激细胞间的组织，连接和通信的工程组织。在这个项目中，一个先进的集成显微镜能够同时光学相干和多光子显微镜，光学相干弹性成像是用来独特地可视化的结构和功能的细胞在3-D工程皮肤结构的关系，并测量不断变化的生物力学性能。第二，本项目调查和纵向图像的3-D工程皮肤结构的生长在不同的微地形基板。这将为深入了解真皮-表皮交界处对角质形成细胞和成纤维细胞的机械影响提供基础。第三，机械刺激对这些结构的影响将进行调查，定义如何不同的刺激影响的3-D细胞动力学和组织随着时间的推移。这样的刺激作用将提供更生理相关的培养条件。最后，该项目将纵向成像皮肤结构移植到宿主临床前模型后的体内细胞和组织反应，为我们理解工程组织移植物与生物宿主的界面做出重大贡献。

1033906BoppartDespite important advances within the past decade, current tissue engineered skin equivalents represent little more than a fragile, short-term dressing for patients that need viable skin replacements. The major weakness in current skin equivalents is that the constituent cells are cultured and applied under conditions that are very different from that of natural skin. It is believed that this is principally due to our limited understanding of the roles that 3-D scaffold topography and mechanical stimuli have on the intercellular organization, connectivity, and communication of engineered tissues. In this project an advanced integrated microscope capable of simultaneous optical coherence and multi-photon microscopy, and optical coherence elastography is utilized to uniquely visualize the structural and functional relationships of cells within 3-D engineered skin constructs, and measure the evolving biomechanical properties. Second, this project investigates and longitudinally images in 3-D the growth of engineered skin constructs on varying microtopographic substrates. This will provide fundamental insight into the mechanical influences at the dermal-epidermal junction on the keratinocytes and fibroblasts. Third, the effects of mechanical stimuli on these constructs will be investigated, defining how varying stimuli affect the 3-D cell dynamics and tissue organization over time. such stimulation effects will provide a more physiologically-relevant culture condition. Finally, this project will longitudinally image the cell and tissue responses in vivo following the grafting of the skin constructs to host pre-clinical models, contributing significantly to our understanding of how engineered tissue grafts interface with biological hosts.