Modelling Medial Degenerative Features by Enzymatic Digestion to Evaluate Disease-Relevant Structure-Function Relationships in the Thoracic Aorta

Abstract

BackgroundAortic microstructure-function relationships and the pathophysiology of how medial degeneration leads to aortic dissection remain poorly defined. We aimed to determine how degeneration of individual components of the extracellular matrix (ECM), namely elastin, collagen, and proteoglycans, influence biomechanical properties of aortic tissue through an improved, disease-motivated enzymatic digestion framework. MethodsPorcine aortic tissue was sectioned into 200 {micro}m thick samples in the media, and progressively digested with elastase or collagenase for selective degradation of these ECM components. Full thickness human aortic tissues were treated with chondroitinase, hyaluronidase, and heparinase to completely remove proteoglycans. Biomechanical characterization was performed using planar biaxial tensile testing, from which low- and high-strain modulus, transition-zone behaviour, strain-energy density, and energy loss were derived. Degree of elastin fiber degradation was analyzed using two photon excitation fluorescence imaging. Analysis of collagen degradation was performed using picrosirius red staining under brightfield and polarized light. Alcian blue staining was used to evaluate proteoglycan content. ResultsInduced fragmentation and disorganization of elastin fibers reduced low-strain load bearing capacity, evidenced by reduced low-strain modulus, strain-energy density, and transition zone stress, along with reduced energy loss. Targeted collagen disorganization similarly reduced strain-energy density and decreased strain at the onset of transition, consistent with premature collagen recruitment, and was accompanied by reductions in high strain modulus and energy loss with increasing collagen degradation. Proteoglycan removal decreased energy loss and was found to modulate low- and high-strain behaviour, including reduced strain-energy density and strain at onset of transition, and increased high strain modulus. ConclusionsThrough targeted modelling of ECM degenerative features on aortic tissue mechanics, we have identified distinct disease-associated biomechanical roles for major matrix constituents, with overlapping effects. These findings delineate mechanical consequences of component-specific matrix degeneration while underscoring the complex, multifactorial nature of structure-function relationships in aortic disease.