Quantitative measurement of the distribution and alignment of collagen fibers in unfixed aortic tissues.

Determination of the local amount and direction of collagen fibers during deformation is crucial for an understanding of the mechanical behavior of aortic tissues. Since most conventional methods cannot be used for this purpose, we propose a method to quantify the local amount and direction of fibers by simply measuring the optical properties of the specimen. After confirming the linear correlation between the retardance and thickness of sections of porcine thoracic aortas (PTAs) ranging from 15 to 300 μm, we investigated the effects of their structural components, i.e., smooth muscle cells (SMCs), elastin and collagen, on the retardance of whole tissues. Decellularization of SMCs did not change the retardance of PTA sections significantly. Patterns in autofluorescent and immunofluorescent images of elastin purified from bovine nuchal ligaments did not match those in retardance images. Images of collagen in PTA sections stained with picrosirius red were similar to corresponding retardance images. The slow axis azimuth corresponded to the circumferential direction of the aorta. Results indicate that collagen in aortas can be quantified by measuring the retardance and slow axis azimuth of whole aortic tissues. Application of this technique to PTAs showed that retardance was higher in dorsal and distal regions than ventral and proximal regions, respectively, indicating that the aortas contain more collagen in distal and dorsal regions than proximal and ventral regions, respectively. Both results were in accordance with previous findings. Measurement of retardance is useful to quantify the amount of collagen in unfixed aortas.