Pregnancy-induced adaptations in intramuscular extracellular matrix of rat pelvic floor muscles

Marianna Alperin, Timothy Kaddis, Rajeswari Pichika, Mary C. Esparza, Richard L. Lieber

Birth trauma to pelvic floor muscles is a major risk factor for pelvic floor disorders. Intramuscular extracellular matrix determines muscle stiffness, supports contractile component, and shields myofibers from mechanical strain. Our goal was to determine whether pregnancy alters extracellular matrix mechanical and biochemical properties in a rat model, which may provide insights into the pathogenesis of pelvic floor muscle birth injury. To examine whether these effects were unique to pelvic floor muscles, we also studied a hind limb muscle. Passive mechanical properties of Coccygeus, Iliocaudalis, Pubocaudalis, and Tibialis Anterior were compared among 3-month old Sprague-Dawley virgin, late-pregnant, and postpartum rats. Muscle tangent stiffness was calculated as the slope of the stress-sarcomere length curve between 2.5 and 4.0 μm, obtained from a stress-relaxation protocol at a bundle level. Elastin and collagen isoform concentrations were quantified using enzyme-linked immunosorbent assay. Enzymatic and glycosylated collagen crosslinks were determined by high-performance liquid chromatography. Data were compared using repeated measures two-way analysis of variance with Tukey's post-hoc testing. Correlations between mechanical and biochemical parameters were assessed by linear regressions. Significance was set to p < 0.05. Results are reported as mean ± standard error of mean. Pregnancy significantly increased stiffness in coccygeus (P<0.05) and pubocaudalis (P<0.0001) relative to virgin controls, with no change in iliocaudalis. Postpartum, pelvic floor muscle stiffness did not differ from virgins, (P>0.3). A substantial increase in collagen V in in coccygeus and pubocaudalis was observed in late-pregnant, compared to virgin animals, (P<0.001). Enzymatic crosslinks decreased in coccygeus (P<0.0001) and pubocaudalis (P<0.02) in pregnancy, while glycosylated crosslinks were significantly elevated in late-pregnant rats in all pelvic floor muscles (P<0.05). Correlations between muscle stiffness and biochemical parameters were inconsistent. In contrast to the changes observed in pelvic floor muscles, the tibialis anterior was unaltered by pregnancy. In contrast to other pelvic tissues, pelvic floor muscle stiffness increased in pregnancy, returning to pre-pregnancy state postpartum. This adaptation may shield myofibers from excessive mechanical strain during parturition. Biochemical alterations in pelvic floor muscle extracellular matrix due to pregnancy include increase in collagen V and a differential response in enzymatic vs. glycosylated collagen crosslinks. The relationships between pelvic floor muscle biochemical and mechanical parameters remain unclear.