Microstructure and mechanics of the bovine trachea: Layer specific investigations through SHG imaging and biaxial testing

The trachea is a complex tissue made up of hyaline cartilage, fibrous tissue, and muscle fibers. Currently, the knowledge of microscopic structural organization of these components and their role in determining the tissue's mechanical response is very limited. The purpose of this study is to provide data on the microstructure of the tracheal components and its influence on tissue's mechanical response. Five bovine tracheae were used in this study. Adventitia, cartilage, mucosa/submucosa, and trachealis muscle layers were methodically cut out from the whole tissue. Second-harmonic generation(SHG) via multi-photon microscopy (MPM) enabled imaging of collagen fibers and muscle fibers. Simultaneously, a planar biaxial test rig was used to record the mechanical behavior of each layer. In total 60 samples were tested and analyzed. Fiber architecture in the adventitia and mucosa/submucosa layer showed high degree of anisotropy with the mean fiber angle varying from sample to sample. The trachealis muscle displayed neat layers of fibers organized in the longitudinal direction. The cartilage also displayed a structure of thick mesh-work of collagen type II organized predominantly towards the circumferential direction. Further, mechanical testing demonstrated the anisotropic nature of the tissue components. The cartilage was identified as the stiffest component for strain level < 20% and hence the primary load bearing component. The other three layers displayed a non-linear mechanical response which could be explained by the structure and organization of their fibers. This study is useful in enhancing the utilization of structurally motivated material models for predicting tracheal overall mechanical response.