Integrating structural heterogeneity, fiber orientation, and recruitment in multiscale ECM mechanics

Extracellular matrix (ECM) plays critical roles in establishing tissue structure-function relationships and controlling cell fate. However, the mechanisms by which ECM mechanics influence cell and tissue behavior remain to be elucidated since the events associated with this process span length scales from the tissue to molecular level. Entirely new methods are needed in order to better understand the multiscale mechanics of ECM. In this study, a multiscale experimental approach was established by integrating Optical Magnetic Twisting Cytometry (OMTC) with a biaxial tensile tester to study the microscopic (local) ECM mechanical properties under controlled tissue-level (global) loading. Adventitial layer of porcine thoracic artery was used as a collagen-based ECM. Multiphoton microscopy imaging was performed to capture the changes in ECM fiber structure during biaxial deformation. As visualized from multiphoton microscopy images, biaxial stretch induces gradual fiber straightening and the fiber families become evident at higher stretch levels. The OMTC measurements show that the local apparent storage and loss modulus increases with the global biaxial stretch, however there exists a complex interplay among local ECM mechanical properties, ECM structural heterogeneity, and fiber distribution and engagement. The phase lag does not change significantly with global biaxial stretch. Our results also show a much faster increase in global tissue tangent modulus compared to the local apparent complex modulus with biaxial stretch, indicating the scale dependency of ECM mechanics.