Marfan syndrome (MFS) is caused by mutation of the FBN1 gene encoding the extracellular matrix (ECM) protein fibrillin-1 that forms elastic microfibrils. This study aims to characterize the micromechanics of aortic and lung tissues from wild type (WT) and FBN1-deficient mutant (MT) mice to identify biophysical determinants of cardiopulmonary disease in MFS. For testing by atomic force microscopy (AFM), full-thickness sections of ascending aorta were dissected from young (0.5-mo) and adult (2-mo) WT and MT mice, mounted flat with the lumen facing up, and indented with a 15-μm spherical probe at 3-5 sites per sample from 1-4 animals per condition. Indentation force-depth curves were analyzed using a hybrid theoretical and finite element analysis to determine the elastic modulus of intima (Eint) and media (Emed) layers. The Kolmogorov-Smirnov test was performed for pairwise statistical analysis; p<0.008 was considered significant based on the Bonferroni correction. Emed at 0.5-mo showed no difference between WT (10.1±2.7kPa, n=8; mean±SEM for n-curves) and MT (7.5±0.6kPa, n=80). Emed increased with age (p<0.0006), but was significantly softer in MT (21.3±1.4kPa, n=112) vs WT (37.4±2.2kPa, n=65, p<0.0001). Eint also increased with age (p<0.003) from ∼2kPa to ∼5-10kPa, but showed no significant difference between WT and MT at matched ages. Similar nano-indentation studies, using a pyramidal probe on lung parenchyma isolated from 2-mo WT and MT mice, showed Elung for MT (1.7±0.1 kPa, n=92) was significantly softer than WT (4.8±0.3kPa, n=91, p<0.0001). In conclusion, AFM revealed age-dependent softening of micro-elastic modulus in elastin-rich tissues including lung parenchyma and aortic media, but not in the aortic intima. In contrast to macro-scale measurements of aortic stiffening in MFS, these micro-scale AFM findings appear consistent with histological observations of local disruption of ECM microstructure in MFS.
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