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

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替代品对于需要可行的皮肤置换的患者来说，只不过是一种易碎的短期敷料。目前皮肤替代物的主要缺点是组成细胞的培养和应用条件与自然皮肤非常不同。据认为，这主要是由于我们对三维支架的地形和机械刺激在工程组织的细胞间组织、连通性和交流中所起的作用了解有限。在这个项目中，一个先进的集成显微镜，能够同时进行光学相干和多光子显微镜，以及光学相干弹性成像，被用来独特地可视化三维工程化皮肤构造中细胞的结构和功能关系，并测量不断演变的生物力学特性。其次，该项目研究并纵向成像在不同微地形基质上的工程化皮肤结构的生长情况。这将从根本上深入了解真皮-表皮交界处对角质形成细胞和成纤维细胞的机械影响。第三，将研究机械刺激对这些结构的影响，确定不同的刺激如何随时间影响3-D细胞动力学和组织结构。这样的刺激效果将提供更具生理相关性的培养条件。最后，该项目将纵向成像移植到宿主临床前模型的皮肤构建物后体内的细胞和组织反应，对我们理解工程化组织移植物如何与生物宿主接口有重要贡献。

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.