Biaxial Mechanical Assessment of the Murine Vaginal Wall Using Extension-Inflation Testing

Progress toward understanding the underlying mechanisms of pelvic organ prolapse (POP) is limited, in part, due to a lack of information on the biomechanical properties and microstructural composition of the vaginal wall. Compromised vaginal wall integrity is thought to contribute to pelvic floor disorders; however, normal structure-function relationships within the vaginal wall are not fully understood. In addition to the information produced from uniaxial testing, biaxial extension-inflation tests performed over a range of physiological values could provide additional insights into vaginal wall mechanical behavior (i.e., axial coupling and anisotropy), while preserving in vivo tissue geometry. Thus, we present experimental methods of assessing murine vaginal wall biaxial mechanical properties using extension-inflation protocols. Geometrically intact vaginal samples taken from 16 female C57BL/6 mice underwent pressure-diameter and force-length preconditioning and testing within a pressure-myograph device. A bilinear curve fit was applied to the local stress-stretch data to quantify the transition stress and stretch as well as the toe- and linear-region moduli. The murine vaginal wall demonstrated a nonlinear response resembling that of other soft tissues, and evaluation of bilinear curve fits suggests that the vagina exhibits pseudoelasticity, axial coupling, and anisotropy. The protocols developed herein permit quantification of biaxial tissue properties. These methods can be utilized in future studies in order to assess evolving structure-function relationships with respect to aging, the onset of prolapse, and response to potential clinical interventions.