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大进展，但目前的组织工程皮肤等效物对于需要可行的皮肤替代品的患者来说，只不过是一种脆弱的短期敷料。目前的皮肤等效物的主要缺点是组成细胞的培养和应用条件与天然皮肤非常不同。据信，这主要是由于我们对三维支架地形和机械刺激对工程组织的细胞间组织、连通性和通信的作用的理解有限。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，光学相干弹性成像被用来独特地可视化三维工程皮肤结构中细胞的结构和功能关系，并测量不断变化的生物力学特性。其次，该项目研究了工程皮肤结构在不同微地形基质上生长的三维纵向图像。这将对真皮-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本的见解。第三，将研究机械刺激对这些结构的影响，定义不同的刺激如何随时间影响3-D细胞动力学和组织组织。这种刺激效应将提供一个更符合生理的培养条件。最后，该项目将纵向成像细胞和组织在体内的反应后，移植皮肤结构宿主临床前模型，有助于我们对工程组织移植物如何与生物宿主界面的理解。

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.