Micromechanical heterogeneity of the rat pia-arachnoid complex

Abstract

To better understand the onset of damage occurring in the brain upon traumatic events, it is essential to analyze how external mechanical loads propagate through the skull and meninges and down to the brain cortex. However, despite their crucial role as structural dampers protecting the brain, the mechanical properties and dynamic behavior of the meningeal layers are still poorly understood. Here, we characterized the local mechanical heterogeneity of rat pia-arachnoid complex (PAC) at the microscale via atomic force microscopy (AFM) indentation experiments to understand how microstructural variations at the tissue level can differentially affect load propagation. By coupling AFM mechanical testing with fresh tissue immunofluorescent staining, we could directly observe the effect of specific anatomical features on the local mechanical properties of tissue. We observed a two-fold stiffening of vascularized tissue when compared to non-vascularized PAC (with instantaneous Young's modulus distribution means of 1.32 ± 0.03 kPa and 2.79 ± 0.08 kPa, respectively), and statistically significant differences between regions of low- and high-vimentin density, reflecting trabecular density (with means of 0.67 ± 0.05 kPa and 1.29 ± 0.06 kPa, respectively). No significant differences were observed between cortical and cerebellar PAC. Additionally, by performing force relaxation experiments at the AFM, we identified the characteristic time constant τ₁ of PAC tissue to be in the range of 2.7 ± 1.2 s to 3.1 ± 0.9 s for the different PAC regions analyzed. Taken together, the results presented point at the complex biomechanical nature of the meningeal tissue, and underscore the need to account for its heterogeneity when modeling its behavior into finite element simulations or other computational methods enabling the prediction of load propagation during injury events. STATEMENT OF SIGNIFICANCE: The meningeal layers are pivotal in shielding the brain during injury events, yet the mechanical properties of this complex biological interface are still under investigation. Here, we present the first anatomically-informed micromechanical characterization of mammalian pia-arachnoid complex (PAC). We developed a protocol for the isolation and fresh immunostaining of rat PAC and subjected the tissue to AFM indentation and relaxation experiments, while visualizing the local anatomy via fluorescence microscopy. We found statistically significant variations in regional PAC stiffness according to the degree of vascularization and trabecular cell density, besides identifying the tissue's characteristic relaxation constant. In essence, this study captures the relationship between anatomy and mechanical heterogeneity in a key component of the brain-skull interface for the first time.

Keywords: Atomic force microscopy; Brain mechanics; Meninges; Pia - arachnoid complex; Traumatic brain injury.