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.
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