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1.

Objective

To undertake a comprehensive analysis of the biochemical tissue composition and passive biomechanical properties of ovine vagina and relate this to the histo-architecture at different reproductive stages as part of the establishment of a large preclinical animal model for evaluating regenerative medicine approaches for surgical treatment of pelvic organ prolapse.

Methods

Vaginal tissue was collected from virgin (n = 3), parous (n = 6) and pregnant sheep (n = 6; mean gestation; 132 d; term = 145 d). Tissue histology was analyzed using H+E and Masson''s Trichrome staining. Biochemical analysis of the extracellular matrix proteins used a hydroxyproline assay to quantify total collagen, SDS PAGE to measure collagen III/I+III ratios, dimethylmethylene blue to quantify glycosaminoglycans and amino acid analysis to quantify elastin. Uniaxial tensiometry was used to determine the Young''s modulus, maximum stress and strain, and permanent strain following cyclic loading.

Results

Vaginal tissue of virgin sheep had the lowest total collagen content and permanent strain. Parous tissue had the highest total collagen and lowest elastin content with concomitant high maximum stress. In contrast, pregnant sheep had the highest elastin and lowest collagen contents, and thickest smooth muscle layer, which was associated with low maximum stress and poor dimensional recovery following repetitive loading.

Conclusion

Pregnant ovine vagina was the most extensible, but the weakest tissue, whereas parous and virgin tissues were strong and elastic. Pregnancy had the greatest impact on tissue composition and biomechanical properties, compatible with significant tissue remodeling as demonstrated in other species. Biochemical changes in tissue protein composition coincide with these altered biomechanical properties.  相似文献   

2.

Objective

There are increasing numbers of reports describing human vaginal tissue composition in women with and without pelvic organ prolapse with conflicting results. The aim of this study was to compare ovine and human posterior vaginal tissue in terms of histological and biochemical tissue composition and to assess passive biomechanical properties of ovine vagina to further characterise this animal model for pelvic organ prolapse research.

Study Design

Vaginal tissue was collected from ovariectomised sheep (n = 6) and from postmenopausal women (n = 7) from the proximal, middle and distal thirds. Tissue histology was analyzed using Masson''s Trichrome staining; total collagen was quantified by hydroxyproline assays, collagen III/I+III ratios by delayed reduction SDS PAGE, glycosaminoglycans by dimethylmethylene blue assay, and elastic tissue associated proteins (ETAP) by amino acid analysis. Young''s modulus, maximum stress/strain, and permanent strain following cyclic loading were determined in ovine vagina.

Results

Both sheep and human vaginal tissue showed comparable tissue composition. Ovine vaginal tissue showed significantly higher total collagen and glycosaminoglycan values (p<0.05) nearest the cervix. No significant differences were found along the length of the human vagina for collagen, GAG or ETAP content. The proximal region was the stiffest (Young''s modulus, p<0.05), strongest (maximum stress, p<0.05) compared to distal region, and most elastic (permanent strain).

Conclusion

Sheep tissue composition and mechanical properties showed regional differences along the postmenopausal vaginal wall not apparent in human vagina, although the absolute content of proteins were similar. Knowledge of this baseline variation in the composition and mechanical properties of the vaginal wall will assist future studies using sheep as a model for vaginal surgery.  相似文献   

3.
This study was undertaken to understand elastin's role in the mechanical homeostasis of the arterial wall. The mechanical properties of elastin vary along the aorta, and we hypothesized this maintained a uniform mechanical environment for the elastin, despite regional variation in loading. Elastin's physiological loading was determined by comparing the inflation response of intact and autoclave purified elastin aortas from the proximal and distal thoracic aorta. Elastin's stretch and stress depend on collagen recruitment. Collagen recruitment started in the proximal aorta at systolic pressures (13.3 to 14.6 kPa) and in the distal at sub-diastolic pressures (9.3 to 10.6 kPa). In the proximal aorta collagen did not contribute significantly to the stress or stiffness, indicating that elastin determined the vessel properties. In the distal aorta, the circumferential incremental modulus was 70% higher than in the proximal aorta, half of which (37%) was due to a stiffening of the elastin. Compared to the elastin tissue in the proximal aorta, the distal elastin suffered higher physiological circumferential stretch (29%, P=0.03), circumferential stress (39%, P=0.02), and circumferential stiffness (37%, P=0.006). Elastin's physiological axial stresses were also higher (67%, P=0.003). These findings do not support the hypothesis that the loading on elastin is constant along the aorta as we expected from homeostasis.  相似文献   

4.
Arteries display a nonlinear anisotropic behavior dictated by the elastic properties and structural arrangement of its main constituents, elastin, collagen, and vascular smooth muscle. Elastin provides for structural integrity and for the compliance of the vessel at low pressure, whereas collagen gives the tensile resistance required at high pressures. Based on the model of Zulliger et al. (Zulliger MA, Rachev A, Stergiopulos N. Am J Physiol Heart Circ Physiol 287: H1335-H1343, 2004), which considers the contributions of elastin, collagen, and vascular smooth muscle cells (VSM) in an explicit form, we assessed the effects of enzymatic degradation of elastin on biomechanical properties of rabbit carotids. Pressure-diameter curves were obtained for controls and after elastin degradation, from which elastic and structural properties were derived. Data were fitted into the model of Zulliger et al. to assess elastic constants of elastin and collagen as well as the characteristics of the collagen engagement profile. The arterial segments were also prepared for histology to visualize and quantify elastin and collagen. Elastase treatment leads to a diameter enlargement, suggesting the existence of significant compressive prestresses within the wall. The elastic modulus was more ductile in treated arteries at low circumferential stretches and significantly greater at elevated circumferential stretches. Abrupt collagen fiber recruitment in elastase-treated arteries leads to a much stiffer vessel at high extensions. This change in collagen engagement properties results from structural alterations provoked by the degradation of elastin, suggesting a clear interaction between elastin and collagen, often neglected in previous constituent-based models of the arterial wall.  相似文献   

5.
Contractility of the proximal and distal vaginal wall smooth muscle may play distinct roles in the female sexual response and pelvic support. The goal of this study was to determine whether differences in contractile characteristics of smooth muscle from these regions reside in differences in the expression of isoforms of myosin, the molecular motor for muscle contraction. Adult female Sprague-Dawley rats were killed on the day of estrus, and the vagina was dissected into proximal and distal segments. The Vmax at peak force was greater for tissue strips of the proximal vagina compared with that of distal (P < 0.01), although, at steady state, the Vmax for the muscle strips from the two regions was not different. Furthermore, at steady state, muscle stress was higher (P < 0.001) for distal vaginal strips (n = 5). Consistent with the high Vmax for the proximal vaginal strips, RT-PCR results revealed a higher %SM-B (P < 0.001) in the proximal vagina. A greater expression of SM-B protein (P < 0.001) was also detected by Western blotting (n = 4). Interestingly, there was no regional difference noted in SM-1/SM-2 isoforms (n = 6). The proximal vagina had a higher expression of myosin heavy chain protein (P < 0.01) and a greater percentage of smooth muscle bundles (P < 0.001). The results of this study are the first demonstration of a regional heterogeneity in Vmax and myosin isoform distribution in the vagina wall smooth muscle and confirm that the proximal vaginal smooth muscle exhibits phasic contractile characteristics compared with the distal vaginal smooth muscle, which is tonic.  相似文献   

6.
Aortas from four sheep and three fetal lambs were fixed at physiological pressure in 10% neutral buffered formalin. The regions with branches were serially sectioned in either cross or longitudinal section at 7-micron intervals and stained for elastin with Gomori-aldehyde-fuchsin. A large model of one aortointercostal junction was made from Plexiglas to show that bundles of elastin appeared to be continuous from the aorta into the branch. These bundles were then studied from large photomicrographs of the other junctions. At the intercostals and lumbars, the elastin lamellae ran continuously from the outer third of the media into the branch. There was often an added "pad" of elastin and other acellular material on the flow divider (distal lip). The large muscular branches which arose from the abdominal aorta have much less elastin than the intercostals. In them the aortic elastin appears to merge into a raphe on the proximal and lateral sides of the junction, with a very abrupt transition. A "tongue" of muscle from the branch often penetrated into the media of the aorta distally. Occasionally a small acellular cap was seen on the apex of the flow divider. There were few significant differences between the lambs and the sheep, probably because embryologically the arteries develop very early. The proximal and distal lips of all junctions were easily distinguished from each other, and the small and large branches were also different. We suspect these regions may respond differently to pressure, but we did not test this hypothesis.  相似文献   

7.
Mechanical characterization of living tissues and computer-based simulations related to medical issues, has become increasingly important to improve diagnostic processes and treatments evaluation. This work proposes a link between the mechanical testing and the material model predictions through histological data of vaginal tissue. Histological data was used to link tensile testing experiments with material-dependent parameters; the approach was adequate to capture the nonlinear response of ovine vaginal tissue over a large strain range.The experimental data obtained on a previous study, has two main components: tensile testing and histological analysis of the ovine vaginal tissue. Uniaxial tensile test data and histological data were collected from three sheep groups: virgins, pregnant and parous. The distal part of vaginal wall was selected since it is prone to tears induced by vaginal delivery.The HGO (Holzapfel-Gasser-Ogden) model parameters were fitted using a stochastic approach, namely the Simple Genetic Algorithm (SGA). The SGA was able to fit the experimental data successfully (R2 > 0.986). The dimensionless coefficient ξ, was highly correlated with histological data. The ratio was seen to increase linearly with increasing collagen content.Coefficient ξ brings a new way of interpreting and understanding experimental data; it connects the nonlinear mechanical behaviour (tensile test) with tissue’s morphology (histology). It can be used as an ‘inverse’ (approximate) method to estimate the mechanical properties without direct experimental measurements, through basic histology.In this context, the proposed methodology appears very promising in estimating the response of the tissue via histological information.  相似文献   

8.
In prairie voles (Microtus ochrogaster), most virgin females are infanticidal. To determine the onset of maternal responsiveness, female prairie voles were tested for maternal behavior as virgins and at different times throughout pregnancy. Female voles that were infanticidal as virgins by and large remained infanticidal throughout pregnancy. In contrast, about 30% of voles that were maternal as virgins became infanticidal during pregnancy. To test whether events associated with parturition facilitate the onset of maternal behavior, females had their litters delivered by Caesarean section within a day of expected delivery or were allowed to give birth naturally with sham surgery occurring shortly before or after birth. Females that gave birth naturally were fully maternal and did not attack unrelated pups, but females subjected to artificial delivery remained infanticidal. This suggests that events closely related to parturition are crucial for full development of maternal behavior in female prairie voles.  相似文献   

9.
10.
Elastin and collagen are the principal scleroproteins of the aortic wall, and they largely determine its physical and mechanical properties. During perinatal development of the aorta, elastin and collagen accumulate rapidly, being present as inverse gradients by the time of birth. Elastin is most prevalent in the thoracic aorta, decreasing distally, while collagen shows the opposite trend. The present studies have determined the relative and absolute rates of collagen and elastin synthesis in the porcine aorta between 60 days of fetal development (mid-gestation) and 110 days after birth. Although there was measurable elastin synthesis in the upper thoracic aorta at the earliest time evaluated, there was a fourfold increase in relative elastin synthesis (from 4 to 16% of total protein synthesis) between 60 fetal days and birth. Elastin synthesis was maximal in successively distal segments between 1 and 3 weeks after birth. Relative collagen synthesis progressively increased in distal aortic regions between 90 fetal days and 60 days postpartum. Greater than twofold increases over thoracic levels were measured. Both elastin and collagen synthesis largely subsided by 110 days of development. When expressed as absolute rates of protein synthesis, these scleroproteins were maximally expressed in the first 3 postnatal weeks. Elastin mRNA levels were determined with a cloned sheep gene fragment by molecular hybridization. Gradients of elastin message were present at 60 fetal days and at 4 and 14 days after birth, elastin mRNA levels being maximal in the upper thoracic aorta at 14 days after birth. The differentiation of the aortic wall thus follows discrete patterns of phenotypic change which may be coupled to the rheologic stresses accompanying development of the circulatory system.  相似文献   

11.
In humans, multiparity (repeated pregnancy) is associated with increased risk of cardiovascular disease. In rats, multiparity increases the pressor response to phenylephrine and to acute stress, due in part to changes in tone of the splanchnic arterial vasculature. Given that the venous system also changes during pregnancy, we studied the effects of multiparity on venous tone and compliance. Cardiovascular responses to volume loading (2 ml/100 g body wt), and mean circulatory filling pressure (MCFP, an index of venomotor tone) were measured in conscious, repeatedly bred (RB), and age-matched virgin rats. In addition, passive compliance and venous reactivity of isolated mesenteric veins were measured by pressure myography. There was a greater increase in mean arterial pressure after volume loading in RB rats (+7.2 +/- 2.5 mmHg, n = 8) than virgin rats (-1.4 +/- 1.7 mmHg, n = 7) (P < 0.05). The increase in MCFP in response to norepinephrine (NE) was also greater in RB rats [half maximal effective dose (ED(50)) 3.1 +/- 0.5 nmol.kg(-1).min(-1), n = 6] than virgins (ED(50): 12.1 +/- 2.7 nmol.kg(-1).min(-1), n = 6) (P < 0.05). Pressure-induced changes in passive diameter were lower in isolated mesenteric veins from RB rats (29.3 +/- 1.8 microm/mmHg, n = 6) than from virgins (36.9 +/- 1.3 microm/mmHg, n = 6) (P < 0.05). Venous reactivity to NE in isolated veins was also greater in RB rats (EC(50): 2.68 +/- 0.37x10(-8) M, n = 5) than virgins (EC(50): 4.67 +/- 0.93 x 10(-8) M, n = 8). We conclude that repeated pregnancy induces a long-term reduction in splanchnic venous compliance and augments splanchnic venous reactivity and sympathetic tonic control of total body venous tone. This compromises the ability of the capacitance (venous) system to accommodate volume overloads and to buffer changes in cardiac preload.  相似文献   

12.
We used a murine model to assess the evolving biomechanical properties of tissue engineered vascular grafts (TEVGs) implanted in the arterial circulation. The initial polymeric tubular scaffold was fabricated from poly(lactic acid)(PLA) and coated with a 50:50 copolymer of poly(caprolactone) and poly(lactic acid)(P[PC/LA]). Following seeding with syngeneic bone marrow derived mononuclear cells, TEVGs (n=50) were implanted as aortic interposition grafts in wild-type mice and monitored serially using ultrasound. A custom biaxial mechanical testing device was used to quantify the in vitro circumferential and axial mechanical properties of grafts explanted at 3 or 7 months. At both times, TEVGs were much stiffer than native tissue in both directions. Repeated mechanical testing of some TEVGs treated with elastase or collagenase suggested that elastin did not contribute significantly to the overall stiffness whereas collagen did contribute. Traditional histology and immunostaining revealed smooth muscle cell layers, significant collagen deposition, and increasing elastin production in addition to considerable scaffold at both 3 and 7 months, which likely dominated the high stiffness seen in mechanical testing. These results suggest that PLA has inadequate in vivo degradation, which impairs cell-mediated development of vascular neotissue having properties closer to native arteries. Assessing contributions of individual components, such as elastin and collagen, to the developing neovessel is needed to guide computational modeling that may help to optimize the design of the TEVG.  相似文献   

13.

Background/Aims

The pacemaker mechanisms activating phasic contractions of vaginal and cervical smooth muscle remain poorly understood. Here, we investigate properties of pacemaking in vaginal and cervical tissues by determining whether: 1) functional pacemaking is dependent on the phase of the estrus cycle or pregnancy; 2) pacemaking involves Ca2+ release from sarcoplasmic/endoplasmic reticulum Ca2+-ATPase (SERCA) -dependent intracellular Ca2+ stores; and 3) c-Kit and/or vimentin immunoreactive ICs have a role in pacemaking.

Methodology/Principal Findings

Vaginal and cervical contractions were measured in vitro, as was the distribution of c-Kit and vimentin positive interstitial cells (ICs). Cervical smooth muscle was spontaneously active in estrus and metestrus but quiescent during proestrus and diestrus. Vaginal smooth muscle was normally quiescent but exhibited phasic contractions in the presence of oxytocin or the K+ channel blocker tetraethylammonium (TEA) chloride. Spontaneous contractions in the cervix and TEA-induced phasic contractions in the vagina persisted in the presence of cyclopiazonic acid (CPA), a blocker of the SERCA that refills intracellular SR Ca2+ stores, but were inhibited in low Ca2+ solution or in the presence of nifedipine, an inhibitor of L-type Ca2+channels. ICs were found in small numbers in the mouse cervix but not in the vagina.

Conclusions/Significance

Cervical smooth muscle strips taken from mice in estrus, metestrus or late pregnancy were generally spontaneously active. Vaginal smooth muscle strips were normally quiescent but could be induced to exhibit phasic contractions independent on phase of the estrus cycle or late pregnancy. Spontaneous cervical or TEA-induced vaginal phasic contractions were not mediated by ICs or intracellular Ca2+ stores. Given that vaginal smooth muscle is normally quiescent then it is likely that increases in hormones such as oxytocin, as might occur through sexual stimulation, enhance the effectiveness of such pacemaking until phasic contractile activity emerges.  相似文献   

14.
Information on the passive biomechanical properties of two-layered esophagus is still limited, although this would enhance our understanding of its physiology/pathophysiology and help to address problems in surgery, medical-device applications, and for the optimal design of prostheses. In this study, rabbit esophagi were excised and dissected into mucosa–submucosa and muscle layers that were submitted to histological quantification of elastin and collagen content and orientation, as well as to inflation-extension testing and geometrical analysis, i.e. delineation of the zero-stress state serving as a reference configuration for biomechanical analysis. The pressure–radius data of both layers displayed a monotonically rising slope with inflating pressure, unlike the sigma shape characterizing elastin-rich tissues, for which biphasic constitutive models were initially postulated. Three phenomenological expressions of strain-energy function (SEF), commonly appearing in soft-tissue biomechanics literature, were used in an attempt to model the pseudoelastic response of esophageal tissue, namely the exponential Fung-type SEF, and the combined neo-Hookean (isotropic) or quadratic (anisotropic) and exponential Fung-type SEF. Accurate fits were attained for the pressure–radius–force data, spanning a wide range of longitudinal stretch ratios, when using the exponential form; the biphasic SEFs failed to generate improved fits, being also over-parameterized. According to the calculated material parameters, mucosa–submucosa was stiffer than muscle in both directions, justified by our histological observation of increased collagen content in that layer, and tissue was stiffer longitudinally, substantiated by the increased elastin and collagen contents and their preferential alignment towards that direction. Our results demonstrate that the passive response of esophagus is best modeled with an exponential Fung-type SEF.  相似文献   

15.
16.
First, we report the results of the longest‐known field study (9 years) to examine the effects of elevated carbon dioxide (CO2) on leaf miner densities in a scrub‐oak community at Kennedy Space Center, Florida. Here, the densities of all leaf miner species (6) on all host species (3) were lower in every year in elevated CO2 than they were in ambient CO2. Second, meta‐analyses were used to review the effects of elevated CO2 on both plants (n=59 studies) and herbivores (n=75 studies). The log of the response ratio was chosen as the metric to calculate effect sizes. Results showed that elevated CO2 significantly decreased herbivore abundance (−21.6%), increased relative consumption rates (+16.5%), development time (+3.87%) and total consumption (+9.2%), and significantly decreased relative growth rate (−8.3%), conversion efficiency (−19.9%) and pupal weight (−5.03%). No significant differences were observed among herbivore guilds. Host plants growing under enriched CO2 environments exhibited significantly larger biomass (+38.4%), increased C/N ratio (+26.57%), and decreased nitrogen concentration (−16.4%), as well as increased concentrations of tannins (+29.9%) and other phenolics. Effects of changes on plant primary and secondary chemistry due to elevated CO2 and consequences for herbivore growth and development are discussed.  相似文献   

17.
One of the largest restoration programs in the world, the Comprehensive Everglades Restoration Plan (CERP) aims to restore freshwater inputs to Everglades wetlands and the Florida Bay estuary. This study predicted how the Florida Bay ecosystem may respond to hydrological restoration from CERP within the context of contemporary projected impacts of sea-level rise (SLR) and increased future temperatures. A spatial–temporal dynamic model (Ecospace) was used to develop a spatiotemporal food web model incorporating environmental drivers of salinity, salinity variation, temperature, depth, distance to mangrove, and seagrass abundance and was used to predict responses of biomass, fisheries catch, and ecosystem resilience between current and future conditions. Changes in biomass between the current and future scenario suggest a suite of winners and losers, with many estuarine species increasing in both total biomass and spatial distribution. Notable biomass increases were predicted for important forage species, including bay anchovy (+32%), hardhead halfbeak (+19%), and pinfish (+31%), while decreases were predicted in mullet (−88%), clupeids (−55%), hardhead silverside (−15%), mojarras (−117%), and Portunid crabs (−16%). Increases in sportfish biomass included the angler-preferred spotted seatrout (+9%), red drum (+10%), and gray snapper (+8%), while decreases included sheepshead (−40%), Atlantic tarpon (−73%), and common snook (−507%). Ecosystem resilience and fisheries catch of angler-preferred species were predicted to improve in the future scenario in total, although a localized decline in resilience predicted for the Central Region may warrant further attention. Our results suggest the Florida Bay ecosystem is likely to achieve restoration benefits in spite of, and in some cases facilitated by, the projected future impacts from climate change due to the system's shallow depth and detrital dominance. The incorporation of climate impacts into long-term restoration planning using ecosystem modeling in similar systems facing unknown futures of SLR, warming seas, and shifting species distributions is recommended.  相似文献   

18.
Understanding the mechanical environment of each component within the arterial wall is fundamental for understanding vascular growth and remodelling and for engineering artificial vascular conduits. We have investigated the mechanical status of arterial elastin by measuring the circumferential mechanical properties of purified elastin as function of position along the descending thoracic aorta of the pig. The tensile circumferential secant modulus, E(sec), measured in uniaxial mechanical tests, increased 30% (P<0.001), from a value of 0.88 MPa in the proximal tissue near the aortic arch to 1.14 MPa in the distal tissue near the diaphragm, indicating the stiffness of the elastin sample increased with position. Breaking stress was 54% higher in the distal tissue compared to the proximal (P<0.001), but the breaking stretch ratio did not change. E(sec) correlated with the ratio of radius to wall thickness measured in the no load state, r(nl)/h(nl), suggesting that the rise in stiffness was linked to ring morphology. The higher stiffness and strength of the distal tissue might be explained by a higher proportion of circumferentially oriented fibres in the distal tissue, which would indicate that the elastin meshwork in the thoracic aorta may become progressively anisotropic with distance from the heart. The ratio r(nl)/(h(nl)E (sec))rose only 7%, which suggests that the in vivo circumferential strain on the elastin may be constant along the pig thoracic aorta. The positional variation in elastin's properties should be taken into account in mechanical studies on purified elastin and in mathematical models of aorta mechanics.  相似文献   

19.
The leg tendons of certain avian species normally calcify. The gastrocnemius, or Achilles, tendon of the domestic turkey, Meleagris gallopavo, is one such example. Its structure and biomechanical properties have been studied to model the adaptive nature of this tendon to external forces, including the means by which mineral deposition occurs and the functional role mineralization may play in this tissue. Structurally, the distal rounded, thick gastrocnemius bifurcates into two smaller proximal segments that mineralize with time. Mineral deposition occurs at or near the bifurcation, proceeding in a distal-to-proximal direction along the segments toward caudal and medial muscle insertions of the bird hip. Mineral formation appears mediated first by extracellular matrix vesicles and later by type I collagen fibrils. Biomechanical analyses indicate lower tensile strength and moduli for the thick distal gastrocnemius compared to narrow, fan-shaped proximal segments. Tendon mineralization here appears to be strain-induced, the muscle forces causing matrix deformation leading conceptually to calcium binding through the exposure of charged groups on collagen, release of sequestered calcium by proteoglycans, and increased diffusion. Functionally, the mineralized tendons limit further tendon deformation, reduce tendon strain at a given stress, and provide greater load-bearing capacity to the tissue. They also serve as important and efficient elastic energy storage reservoirs, increasing the amount of stored elastic energy by preventing flexible type I collagen regions from stretching and preserving muscle energy during locomotion of the animals.  相似文献   

20.

Background  

During normal pregnancy the cervix has a load bearing function. The cervical tissue consists mainly of an extracellular matrix (ECM) rich in collagen; important for the biomechanical properties. The aim of the present study was to evaluate how the biomechanical strength of samples from the distal cervix is associated with collagen content in relation to age and parity. This study demonstrates a method to investigate cervical tissue from women who still have their uterus in situ.  相似文献   

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