Experimental Characterization of Adventitial Collagen Fiber Kinematics Using Second-Harmonic Generation Imaging Microscopy: Similarities and Differences Across Arteries, Species and Testing Conditions

Fibrous collagen networks are well known to play a central role in the passive biomechanical response of soft connective tissues to applied loads. In the current chapter we focus on vascular tissues and share our extensive experience in coupling mechanical loading and multi-photon imaging to investigate, across arteries, species and testing conditions, how collagen fibers move in response to mechanical loading. More specifically, we assess the deformations of collagen networks in rabbit, porcine or human arteries under different loading scenarios: uniaxial tension on flat samples, tension-inflation on tubular samples, bulge inflation on flat samples. We always observe that collagen fibers exhibit a wavy or crimped shape in load-free conditions, and tend to uncrimp when loads are applied, engaging sequentially to become the main load-bearing component. This sequential engagement, which is responsible for the nonlinear mechanical behaviour, is essential for an artery to function normally and appears to be less pronounced for arteries in elderly and aneurysmal patients. Although uncrimping of collagen fibers is a universal mechanism, we also observe large fiber rotations specific to tensile loading, with significant realignment along the loading axis. A unified approach is proposed to compare observations and quantitative analyses as the type of image processing may affect significantly the estimation of collagen fiber deformations. In summary, this chapter makes an important review of the basic roles of arterial microstructure and its deformations on the global mechanical response. Eventually, directions for future studies combining mechanical loading and multi-photon imaging are suggested, with the aim of addressing open questions related to tissue adaptation and rupture.