Characterizing Tissue Development Under Biaxial Loading

描述（由申请人提供）：我们建议设计和建造一个定制的培养室系统，该系统可以提供可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）的显微镜台上。一个多学科团队已经组建，整合了机械生物学、非线性光学和组织工程方面的专业知识。培养室系统将是第一个此类系统，用于组织响应受控机械载荷的纵向研究，使用NLOM来评估微观组织和基质含量，并通过应力应变测量来表征单个组织样本的整体机械性能。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，并描绘负责将细胞表达谱转化为整体组织反应的生物事件变得可行。在为期两年的项目期内，我们建议建立这种新型培养室系统，并建立使用NLOM获得细胞外基质参数定量测量的方案。这些实验方案旨在监测组织微观结构和整体力学性能在三周内的演变变化。这些定量的NLOM测量将与完全相同组织标本的整体生物力学特性相关联，并将为我们正在发展的组织生长和重塑的数学理论提供实验参数。这个新颖的实验平台将为机械生物学的新颖和创新研究奠定基础。

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.