Characterizing Tissue Development Under Biaxial Loading

描述（由申请人提供）：我们建议设计和构建一个定制的培养室系统，该系统可以提供受控的机械组织培养环境，并安装在用于非线性光学显微镜（NLOM）的显微镜载物台上。一个多学科的团队已经组装，整合在机械生物学，非线性光学和组织工程的专业知识。该培养室系统将是同类系统中的第一个，用于组织对受控机械载荷的响应的纵向研究，使用NLOM评估微观结构组织和基质含量，并测量应力-应变，以表征单个组织标本的整体机械性能。通过将生物力学测量与成分特异性、非破坏性、活体成像相结合，可以直接研究基本的结构-功能关系，并且描绘负责将细胞表达谱转化为大量组织反应的生物事件变得可行。在两年的项目期间，我们建议建立这种新型的培养室系统，并建立使用NLOM获取细胞外基质参数的定量测量的协议。这些实验方案将被设计为监测组织微观结构和体积力学性能在三周内的不断变化。这些定量NLOM测量将与完全相同的组织标本上的体积生物力学特性相关，并将为我们开发的组织生长和重塑的数学理论提供实验参数。这个新的实验平台将为机械生物学的新的和创新的研究奠定基础。

DESCRIPTION (provided by applicant): We propose to design and construct a custom culture chamber system which can provide controlled mechanical tissue culture environments and be mounted on a microscope stage used for nonlinear optical microscopy (NLOM). A multidisciplinary team has been assembled, integrating expertise in mechanobiology, nonlinear optics and tissue engineering. The culture chamber system will be the first of its kind, designed for longitudinal studies of tissue response to controlled mechanical loads using NLOM to evaluate microstructure organization and matrix content and stress-strain measurements to characterize bulk mechanical properties on individual tissue specimens. By combining biomechanical measurements with constituent specific, non-destructive, intravital imaging, fundamental structure-function relationships may be directly studied and delineating biological events responsible for translating cellular expression profiles into bulk tissue response becomes feasible. In the two year project period, we propose to build this novel culture chamber system and establish protocols for acquiring quantitative measures of extracellular matrix parameters using NLOM. These experimental protocols will be designed to monitor evolving changes in tissue microstructure and bulk mechanical properties over a three week period. These quantitative NLOM measurements will be correlated with bulk biomechanical properties on the exact same tissue specimen and will provide experimental parameters for our developing mathematical theory of tissue growth and remodeling. This novel experimental platform will form the basis for novel and innovative studies in mechanobiology.