The Role of Biaxial Loading on Smooth Muscle Contractility in the Nulliparous Murine Cervix.

Throughout the estrus cycle, the extracellular matrix (ECM) and cervical smooth muscle cells (cSMC) coordinate to accomplish normal physiologic function in the non-pregnant cervix. While previous uniaxial experiments provide fundamental knowledge about cervical contractility and biomechanics, the specimen preparation is disruptive to native organ geometry and does not permit simultaneous assessment of circumferential and axial properties. Thus, a need remains to investigate cervical contractility and passive biomechanics within physiologic multiaxial loading. Biaxial inflation-extension experiments overcome these limitations by preserving geometry, ECM-cell interactions, and multiaxially loading the cervix. Utilizing in vivo pressure measurements and inflation-extension testing, this study presented methodology and examined maximum biaxial contractility and biomechanics in the nulliparous murine cervix. The study showed that increased pressure resulted in decreased contractile potential in the circumferential direction, however, axial contractility remained unaffected. Additionally, total change in axial stress ([Formula: see text]) increased significantly (p < 0.05) compared to circumferential stress ([Formula: see text]) with maximum contraction. However, passive stiffness was significantly greater (p < 0.01) in the circumferential direction. Overall, axial cSMC may have a critical function in maintaining cervical homeostasis during normal function. Potentially, a loss of axial contractility in the cervix during pregnancy may result in maladaptive remodeling such as cervical insufficiency.

Pelvic Organ Prolapse: A Review of In Vitro Testing of Pelvic Support Mechanisms.

Background: Pelvic organ prolapse (POP) affects a significant portion of the female population, impacting quality of life and often requiring intervention. The exact cause of prolapse is unknown. Methods: We review some of the current research that focuses on defining the elements involved in POP, with a focus on in vitro testing. Results: Treatment for POP, ranging from physical therapy or pessary use to more invasive surgery, has varying success rates. This variation is, in part, because the pathophysiology of pelvic floor support-and thus dysfunction-is incompletely understood, particularly regarding the structural components and biomechanical properties of tissue. However, researchers are working to identify and quantify the structural and functional dysfunction that may lead to the development of this condition. Conclusion: Given the limited understanding of prolapse development, more research is needed to quantify the microstructure of the pelvic organs and pelvic support structures, with and without prolapse. Identifying biomechanical properties in multiaxial configurations will improve our understanding of pelvic tissue support, as well as our ability to establish predictive models and improve clinical treatment strategies.

Biaxial Basal Tone and Passive Testing of the Murine Reproductive System Using a Pressure Myograph.

The female reproductive organs, specifically the vagina and cervix, are composed of various cellular components and a unique extracellular matrix (ECM). Smooth muscle cells exhibit a contractile function within the vaginal and cervical walls. Depending on the biochemical environment and the mechanical distension of the organ walls, the smooth muscle cells alter the contractile conditions. The contribution of the smooth muscle cells under baseline physiological conditions is classified as a basal tone. More specifically, a basal tone is the baseline partial constriction of smooth muscle cells in the absence of hormonal and neural stimulation. Furthermore, the ECM provides structural support for the organ walls and functions as a reservoir for biochemical cues. These biochemical cues are vital to various organ functions, such as inciting growth and maintaining homeostasis. The ECM of each organ is composed primarily of collagen fibers (mostly collagen types I, III, and V), elastic fibers, and glycosaminoglycans/proteoglycans. The composition and organization of the ECM dictate the mechanical properties of each organ. A change in ECM composition may lead to the development of reproductive pathologies, such as pelvic organ prolapse or premature cervical remodeling. Furthermore, changes in ECM microstructure and stiffness may alter smooth muscle cell activity and phenotype, thus resulting in the loss of the contractile force. In this work, the reported protocols are used to assess the basal tone and passive mechanical properties of the nonpregnant murine vagina and cervix at 4-6 months of age in estrus. The organs were mounted in a commercially available pressure myograph and both pressure-diameter and force-length tests were performed. Sample data and data analysis techniques for the mechanical characterization of the reproductive organs are included. Such information may be useful for constructing mathematical models and rationally designing therapeutic interventions for women's health pathologies.

Biaxial biomechanical properties of the nonpregnant murine cervix and uterus.

From a biomechanical perspective, female reproductive health is an understudied area of research. There is an incomplete understanding of the complex function and interaction between the cervix and uterus. This, in part, is due to the limited research into multiaxial biomechanical functions and geometry of these organs. Knowledge of the biomechanical function and interaction between these organs may elucidate etiologies of conditions such as preterm birth. Therefore, the objective of this study was to quantify the multiaxial biomechanical properties of the murine cervix and uterus using a biaxial testing set-up. To accomplish this, an inflation-extension testing protocol (n = 15) was leveraged to quantify biaxial biomechanical properties while preserving native matrix interactions and geometry. Ultrasound imaging and histology (n = 10) were performed to evaluate regional geometry and microstructure, respectively. Histological analysis identified a statistically significant greater collagen content and significantly smaller smooth muscle content in the cervix as compared to the uterus. No statistically significant differences in elastic fibers were identified. Analysis of bilinear fits revealed a significantly stiffer response from the circumferentially orientated ECM fibers compared to axially orientated fibers in both organs. Bilinear fits and a two-fiber family constitutive model showed that the cervix was significantly less distensible than the uterus. We submit that the regional biaxial information reported in this study aids in establishing an appropriate reference configuration for mathematical models of the uterine-cervical complex. Thus, may aid future work to elucidate the biomechanical mechanisms leading to cervical or uterine conditions.

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.