Sexual Dimorphism in the Architectural Design of Rat and Human Pelvic Floor Muscles.

Skeletal muscle architecture is a strong predictor of in vivo functional capacity and is evaluated in fixed tissues, accommodating the study of human muscles from cadaveric donors. Previous studies evaluating the pelvic floor muscles (PFMs) demonstrated that the rat is the most appropriate small animal model for the study of female PFM architecture, but the rat's suitability for the study of male PFMs is undetermined. We aimed to determine (1) whether PFM architecture exhibits sexual dimorphism in rats or humans, and (2) if the rat is also a suitable animal model for the study of male human PFMs. PFMs were fixed in situ and harvested en bloc from male and female cadaveric donors and 3-month-old male and female Sprague-Dawley rats. Three architectural parameters influenced by species size were used to compare male versus female PFMs within species, while four size-independent measures compared species within sex. All comparisons were made with two-way analysis of variances and Tukey's multiple comparisons tests post hoc. Sarcomere length (rats and humans, p = 0.016 and = 0.002) and normalized fiber length (rats, p < 0.001) were significantly larger in male PFMs. Three of the size-independent measures exhibited similar species trends in both sexes, while the size-independent sarcomere length measure (Ls/Lso) differed between male rats and humans (p < 0.001). Thus, sexual dimorphism is present in rat and human PFM architecture, and the male rat is suitable for studies of human male PFMs.

Novel Application of Photogrammetry to Quantify Fascicle Orientations of Female Cadaveric Pelvic Floor Muscles.

Although critical for understanding and simulating pelvic floor muscle function and pathophysiology, the fascicle arrangements of the coccygeus and levator ani remain mostly undetermined. We performed close-range photogrammetry on cadaveric pelvic floor muscles to robustly quantify surface fascicle orientations. The pelvic floor muscles of 5 female cadavers were exposed through anatomic dissections, removed en bloc, and photographed from every required angle. Overlapping images were mapped onto in silico geometries and muscle fascicles were traced manually. Tangent vectors were calculated along each trace; interpolated to define continuous, 3D vector fields; and projected onto axial and sagittal planes to calculate angles with respect to the pubococcygeal line. Contralateral and ipsilateral pelvic floor muscles were compared within each donor (Kuiper's tests) and using mean values from all donors (William-Watsons tests). Contralateral muscles and all but one ipsilateral muscle pair differed significantly within each donor (p < 0.001). When mean values were considered collectively, no contralateral or ipsilateral statistical differences were found but all muscles compared differed by more than 10° on average. Close-range photogrammetry and subsequent analyses robustly quantified surface fascicle orientations of the pelvic floor muscles. The continuous, 3D vector fields provide data necessary for improving simulations of the female pelvic floor muscles.

Structure-function relationship of the human external anal sphincter.

INTRODUCTION AND HYPOTHESIS: Obstetrical external anal sphincter (EAS) injury and subsequent dysfunction are leading risk factors for female fecal incontinence (FI). Limited knowledge of the EAS structure-function relationship hinders treatment optimization. We directly measured functionally relevant intrinsic parameters of human EAS and tested whether vaginal delivery alters the EAS structure-function relationship. METHODS: Major predictors of in vivo EAS function were compared between specimens procured from vaginally nulliparous (VN, n = 5) and vaginally parous (VP, n = 7) cadaveric donors: operational sarcomere length (Ls), which dictates force-length relationship; physiological cross-sectional area (PCSA), which determines isometric force-generating capacity; fiber length (Lfn), responsible for muscle excursion and contractile velocity; and muscle stiffness. Data were analyzed using unpaired and paired t tests, α < 0.05. Results are presented as mean ± SEM. RESULTS: The VN and VP (median parity 3) groups were similar in age and BMI. No gross anatomical defects were identified. EAS Ls (2.36 ± 0.05 µm) was shorter than the optimal Lso (2.7 µm), at which contractile force is maximal, P = 0.0001. Stiffness was lower at Ls than Lso (5.4 ± 14 kPa/µm vs 35.3 ± 12 kPa/µm, P < 0.0001). This structural design allows active and passive tension to increase with EAS stretching. EAS relatively long Lfn (106 ± 24.8 mm) permits rapid contraction without decreased force, whereas intermediate PCSA (1.3 ± 0.3 cm2) is conducive to maintaining resting tone. All parameters were similar between groups. CONCLUSIONS: This first direct examination of human EAS underscores how EAS intrinsic design matches its intended function. Knowledge of the EAS structure-function relationship is important for understanding the pathogenesis of FI and the optimization of treatments for EAS dysfunction.

Architectural assessment of rhesus macaque pelvic floor muscles: comparison for use as a human model.

INTRODUCTION AND HYPOTHESIS: Animal models are essential to further our understanding of the independent and combined function of human pelvic floor muscles (PFMs), as direct studies in women are limited. To assure suitability of the rhesus macaque (RM), we compared RM and human PFM architecture, the strongest predictor of muscle function. We hypothesized that relative to other models, RM best resembles human PFM. METHODS: Major architectural parameters of cadaveric human coccygeus, iliococcygeus, and pubovisceralis (pubococcygeus + puborectalis) and corresponding RM coccygeus, iliocaudalis, and pubovisceralis (pubovaginalis + pubocaudalis) were compared using 1- and 2-way analysis of variance (ANOVA) with post hoc testing. Architectural difference index (ADI), a combined measure of functionally relevant structural parameters predictive of length-tension, force-generation, and excursional muscle properties was used to compare PFMs across RM, rabbit, rat, and mouse. RESULTS: RM and human PFMs were similar with respect to architecture. However, the magnitude of similarity varied between individual muscles, with the architecture of the most distinct RM PFM, iliocaudalis, being well suited for quadrupedal locomotion. Except for the pubovaginalis, RM PFMs inserted onto caudal vertebrae, analogous to all tailed animals. Comparison of the PFM complex architecture across species revealed the lowest, thus closest to human, ADI for RM (1.9), followed by rat (2.0), mouse (2.6), and rabbit (4.7). CONCLUSIONS: Overall, RM provides the closest architectural representation of human PFM complex among species examined; however, differences between individual PFMs should be taken into consideration. As RM is closely followed by rat with respect to PFM similarity with humans, this less-sentient and substantially cheaper model is a good alternative for PFM studies.

An unembalmed cadaveric preparation for simulating pleural effusion: A pilot study of chest percussion involving medical students.

Cadaveric simulations are an effective way to add clinical context to an anatomy course. In this study, unembalmed (fresh) cadavers were uniquely prepared to simulate pleural effusion to teach chest percussion and review thoracic anatomy. Thirty first-year medical students were assigned to either an intervention (Group A) or control group (Group B). Group A received hands-on training with the cadaveric simulations. They were instructed on how to palpate bony landmarks for identifying the diaphragm and lobes of the lungs, as well as on how to properly perform chest percussion to detect abnormal fluid in the pleural space. Students in Group B practiced on each other. Students in Group A benefited from the training in several ways. They had more confidence in their percussive technique (A = mean 4.3/5.0, B = 2.9/5.0), ability to count the ribs on an intact body (A = mean 4.0/5.0, B = 3.0/5.0), and ability to identify the lobes of the lungs on an intact body (A = mean 3.8/5.0, B = 2.3/5.0). They also demonstrated a greater ability to locate the diaphragm on an intact body (A = 100%, B = 60%) and detect abnormal pleural fluid (A = 93%, B = 53%) with greater confidence (A = mean 3.7/5.0, B = 2.5/5.0). Finally, the hands-on training with the unembalmed cadavers created more excitement around learning in Group A compared with Group B. This study shows that simulating pleural effusion in an unembalmed cadaver is a useful way to enhance anatomy education. Anat Sci Educ 10: 160-169. © 2016 American Association of Anatomists.

Age-related alterations in female obturator internus muscle.

INTRODUCTION AND HYPOTHESIS: Pelvic floor muscle rehabilitation is a widely utilized, but often challenging therapy for pelvic floor disorders, which are prevalent in older women. Regimens involving the use of appendicular muscles, such as the obturator internus (OI), have been developed for strengthening of the levator ani muscle (LAM). However, changes that lead to potential dysfunction of these alternative targets in older women are not well known. We hypothesized that aging negatively impacts OI architecture, the main determinant of muscle function, and intramuscular extracellular matrix (ECM), paralleling age-related alterations in LAM. METHODS: OI and LAM were procured from three groups of female cadaveric donors (five per group): younger (20 - 40 years), middle-aged (41 - 60 years), and older (≥60 years). Architectural predictors of the excursional (fiber length, L f), force-generating (physiological cross-sectional area, PCSA) and sarcomere length (L s) capacity of the muscles, and ECM collagen content (measure of fibrosis) were determined using validated methods. The data were analyzed using one-way ANOVA and Tukey's post-hoc test with a significance level of 0.05, and linear regression. RESULTS: The mean ages of the donors in the three groups were 31.2 ± 2.3 years, 47.6 ± 1.2 years, and 74.6 ± 4.2 years (P < 0.005). The groups did not differ with respect to parity or body mass index (P > 0.5). OI L f and L s were not affected by aging. Age >60 years was associated with a substantial decrease in OI PCSA and increased collagen content (P < 0.05). Reductions in OI and LAM force-generating capacities with age were highly correlated (r 2 = 0.9). CONCLUSIONS: Our findings of age-related decreases in predicted OI force production and fibrosis suggest that these alterations should be taken into consideration, when designing pelvic floor fitness programs for older women.

Architectural design of the pelvic floor is consistent with muscle functional subspecialization.

INTRODUCTION AND HYPOTHESIS: Skeletal muscle architecture is the strongest predictor of a muscle's functional capacity. The purpose of this study was to define the architectural properties of the deep muscles of the female pelvic floor (PFMs) to elucidate their structure-function relationships. METHODS: PFMs coccygeus (C), iliococcygeus (IC), and pubovisceral (PV) were harvested en bloc from ten fixed human cadavers (mean age 85 years, range 55-102). Fundamental architectural parameters of skeletal muscles [physiological cross-sectional area (PCSA), normalized fiber length, and sarcomere length (L(s))] were determined using validated methods. PCSA predicts muscle-force production, and normalized fiber length is related to muscle excursion. These parameters were compared using repeated measures analysis of variance (ANOVA) with post hoc t tests, as appropriate. Significance was set to α = 0.05. RESULTS: PFMs were thinner than expected based on data reported from imaging studies and in vivo palpation. Significant differences in fiber length were observed across PFMs: C = 5.29 ± 0.32 cm, IC = 7.55 ± 0.46 cm, PV = 10.45 ± 0.67 cm (p < 0.001). Average L(s) of all PFMs was short relative to the optimal L(s) of 2.7 µm of other human skeletal muscles: C = 2.05 ± 0.02 µm, IC = 2.02 ± 0.02 µm, PC/PR = 2.07 ± 0.01 µm (p = <0.001 compared with 2.7 µm; p = 0.15 between PFMs, power = 0.46). Average PCSA was very small compared with other human muscles, with no significant difference between individual PFMs: C = 0.71 ± 0.06 cm(2), IC = 0.63 ± 0.04 cm(2), PV = 0.59 ± 0.05 cm(2) (p = 0.21, power = 0.27). Overall, C had shortest fibers, making it a good stabilizer. PV demonstrated the longest fibers, suggesting that it functions to produce large excursions. CONCLUSIONS: PFM design shows individual muscles demonstrating differential architecture, corresponding to specialized function in the pelvic floor.

Human skeletal muscle biochemical diversity.

The molecular components largely responsible for muscle attributes such as passive tension development (titin and collagen), active tension development (myosin heavy chain, MHC) and mechanosensitive signaling (titin) have been well studied in animals but less is known about their roles in humans. The purpose of this study was to perform a comprehensive analysis of titin, collagen and MHC isoform distributions in a large number of human muscles, to search for common themes and trends in the muscular organization of the human body. In this study, 599 biopsies were obtained from six human cadaveric donors (mean age 83 years). Three assays were performed on each biopsy - titin molecular mass determination, hydroxyproline content (a surrogate for collagen content) and MHC isoform distribution. Titin molecular mass was increased in more distal muscles of the upper and lower limbs. This trend was also observed for collagen. Percentage MHC-1 data followed a pattern similar to collagen in muscles of the upper extremity but this trend was reversed in the lower extremity. Titin molecular mass was the best predictor of anatomical region and muscle functional group. On average, human muscles had more slow myosin than other mammals. Also, larger titins were generally associated with faster muscles. These trends suggest that distal muscles should have higher passive tension than proximal ones, and that titin size variability may potentially act to 'tune' the protein's mechanotransduction capability.