Smooth Muscle Organization and Nerves in the Rat Vagina: A First Look Using Tissue Clearing and Immunolabeling.

Smooth muscle fibers within the vagina, as well as the nerve fibers that contribute to their control mechanisms, are important for the maintenance and alteration of vaginal length and tone. Vaginal smooth muscle (VaSM) is typically described as being arranged into two distinct concentric layers: an inner circular muscular layer and an outer longitudinal muscular layer. However, the distribution of VaSM oriented in the longitudinal direction (LD) and circumferential direction (CD) has never been quantified. In this study, tissue clearing and immunohistochemistry were performed so that the VaSM, and surrounding nerves, within whole rat vaginas ([Formula: see text]) could be imaged without tissue sectioning, preserving the three-dimensional architecture of the organs. Using these methods, the vagina was viewed through the full thickness of the muscularis layer, from the distal to the proximal regions. The VaSM orientation in the proximal and distal regions and the VaSM content along the LD and CD were quantified. Additionally, a qualitative assessment of vaginal nerves was performed. When compared using a permuted version of the Watson [Formula: see text] test, the orientation of VaSM in the proximal and distal regions were found to be significantly different in 4 of the 6 imaged rat vaginas ([Formula: see text]). While the distal vagina contained a similar amount of VaSM oriented within [Formula: see text] of the LD and within [Formula: see text] of the CD, the proximal vagina contained significantly more VaSM oriented towards the LD than towards the CD. Nerve fibers were found to be wavy, running both parallel and perpendicular to vascular and non-vascular smooth muscle within the vagina. Micro-structural analyses, like the one conducted here, are necessary to understand the physiological function and pathological changes of the vagina.

Strains induced in the vagina by smooth muscle contractions.

The ability of the vagina to contract gives rise to a set of active mechanical properties that contribute to the complex function of this organ in-vivo. Regional differences in the morphology of the vagina have been long recognized, but the large heterogeneous deformations that the vagina experiences during contractions have never been quantified. Furthermore, there is no consensus regarding differences in contractility along the two primary anatomical directions of the vagina: the longitudinal direction (LD) and the circumferential direction (CD). In this study, square vaginal specimens from healthy virgin rats (n=15) were subjected to isometric planar biaxial tests at four equi-biaxial stretches of 1.0, 1.1, 1.2, and 1.3. Contractions were induced at each stretch by a high concentration potassium solution. The digital image correlation method was used to perform full-field strain measurements during contractions. The vagina was found to undergo significantly higher compressive strains, tensile strains, and contractile forces along the LD than along the CD during contractions. Specifically, when computed over all the applied equi-biaxial stretches, mean (± std. dev.) absolute maximum compressive strains were -(13.43 ± 1.56)% along the LD and -(3.19 ± 0.25)% along the CD, mean absolute maximum tensile strains were (10.92 ± 1.73)% along the LD and (3.62 ± 0.57)% along the CD, and mean maximum contractile forces were 6.24 ± 0.55 mN along the LD and 3.35 ± 0.56 mN along the CD. Moreover, the vaginal tissue appeared to undergo compression in the proximal region and tension in the distal region while kept at constant equi-biaxial stretches. The active mechanical properties of the healthy vagina need to be fully investigated so that detrimental alterations in vaginal contractility, such as those caused by pelvic floor disorders and current treatment strategies, can be prevented. STATEMENT OF SIGNIFICANCE: Contractile forces of the vagina have been measured by several investigators using uniaxial tensile testing methods. Unlike previous studies, in this study planar-biaxial tests of vaginal specimens were performed while the full-field strains of the vagina, as induced by smooth muscle contraction, were measured. The vagina was found to generate significantly larger contractile strains and forces in the longitudinal direction than in the circumferential direction. Knowledge of the contractile mechanics of the healthy vagina is essential to understand the detrimental effects that pelvic organ prolapse and the use of surgical meshes have on the functionality of smooth muscle in the vagina.

Mechanics of Uterosacral Ligaments: Current Knowledge, Existing Gaps, and Future Directions.

The uterosacral ligaments (USLs) are important anatomical structures that support the uterus and apical vagina within the pelvis. As these structures are over-stretched, become weak, and exhibit laxity, pelvic floor disorders such as pelvic organ prolapse occur. Although several surgical procedures to treat pelvic floor disorders are directed toward the USLs, there is still a lot that is unknown about their function. This manuscript presents a review of the current knowledge on the mechanical properties of the USLs. The anatomy, microstructure, and clinical significance of the USLs are first reviewed. Then, the results of published experimental studies on the in vivo and ex vivo, uniaxial and biaxial tensile tests are compiled. Based on the existing findings, research gaps are identified and future research directions are discussed. The purpose of this exhaustive review is to help new researchers navigate scientific literature on the mechanical properties of the USLs. The use of these structures remains very popular in reconstructive surgeries that restore and augment the support of pelvic organs, especially as synthetic surgical mesh implants continue to be highly controversial.

Contractile Properties of Vaginal Tissue.

The vagina is an important organ of the female reproductive system that has been largely understudied in the field of biomechanics. In recent years, some research has been conducted to evaluate the mechanical properties of the vagina, but much has focused on characterizing the passive mechanical properties. Because vaginal contractions play a central role in sexual function, childbirth, and development and treatment of pelvic floor disorders, the active mechanical properties of the vagina must be also quantified. This review surveys and summarizes published experimental studies on the active properties of the vagina including the differences in such properties determined by anatomic regions and orientations, neural pathways, life events such as pregnancy and menopause, pelvic floor disorders such as prolapse, and surgical mesh treatment. Conflicting experimental findings are presented, illustrating the need for further research on the active properties of the vagina. However, consensus currently exists regarding the negative impact of surgical mesh on vaginal contractility. This review also identifies knowledge gaps and future research opportunities, thus proving a firm foundation for novice and experienced researchers in this emerging area of biomechanics and encouraging more activity on women's sexual and reproductive health research.

Anisotropy of the Passive and Active Rat Vagina Under Biaxial Loading.

Pelvic organ prolapse, the descent of the pelvic organs from their normal anatomical position, is a common condition among women that is associated with mechanical alterations of the vaginal wall. In order to characterize the complex mechanical behavior of the vagina, we performed planar biaxial tests of vaginal specimens in both the passive (relaxed) and active (contracted) states. Specimens were isolated from virgin, female Long-Evans rats (n = 16) and simultaneously stretched along the longitudinal direction (LD) and circumferential direction (CD) of the vagina. Tissue contraction was induced by electric field stimulation (EFS) at incrementally increasing values of stretch and, subsequently, by KCl. On average, the vagina was stiffer in the CD than in the LD (p < 0.001). The mean maximum EFS-induced active stress was significantly higher in the CD than in the LD (p < 0.01). On the contrary, the mean KCl-induced active stress was lower in the CD than in the LD (p < 0.01). When comparing the mean maximum EFS-induced active stress to the mean KCl-induced active stress, no differences were found in the CD (p = 0.366) but, in the LD, the mean active stress was much higher in response to the KCl stimulation (p < 0.001). Collectively, these results suggest that the anisotropic behavior of the vaginal tissue is determined not only by collagen and smooth muscle fiber organization but also by the innervation.

Anterior Cervical Discectomy With Fusion Using a Local Source for Cancellous Autograft: A Biomechanical Analysis of Vertebral Body Stability in an Osteopenic Bone Model.

BACKGROUND: Anterior cervical discectomy with fusion is an effective treatment for patients having cervical radiculopathy and myelopathy. To reduce morbidity associated with autograft taken from the iliac crest without sacrificing high fusion rates, a novel technique that harvests bone from the vertebral body adjacent to the operative disc space has been proposed. The effects of square and round bone graft harvest techniques on the mechanical stability of the osteopenic donor vertebrae are unknown. We analyzed the biomechanical implications of the technique by subjecting osteopenic models to uniaxial compression to compare yield strengths of surgically altered and unaltered specimens. METHODS: Biomechanical grade polyurethane foam was cut into 60 different 12 mm × 17 mm × 20 mm blocks. The foam had a density of 10 pounds per cubic foot, simulating osteoporotic bone. Rectangular prism (4 mm × 4 mm × 6 mm) and cylindrical cores (r = 2 mm, h = 8 mm) were removed from 20 blocks per group. Twenty samples were left intact as a control group. Anterior plate screws were applied to the models and a Polyether ether ketone (PEEK) interbody spacer was placed on top. Samples underwent uniaxial compression at 0.1 mm/s until mechanical failure. Points of structural failure were determined using a 0.1% offset on a force-displacement curve and compared to determine the reductions in compressive strength. RESULTS: The mean force eliciting structural failure for intact samples was 450.6 N. Average failure forces for rectangular prisms and cylindrical cores removed were 383.2 and 395.4 N, respectively. Removal of a rectangular prismatic core of the necessary volume resulted in a 15.0% reduction in compressive strength, while removal of a cylindrical core of comparable volume facilitated a reduction of 12.2%. CONCLUSION: Local autograft harvested from adjacent vertebrae reduces morbidity associated with a second surgical site while minimally reducing the compressive strength of the donor vertebra in an osteopenic model, lending credence to the efficacy of this technique in elderly patient populations.