elastin and Pelvic-Organ-Prolapse

Pelvic organ prolapse is a frequent health problem in women, encountered worldwide, its physiopathology being still incompletely understood. The integrity of the pelvic-supportive structures is a key element that prevents the prolapse of the pelvic organs. Numerous researchers have underlined the role of connective tissue molecular changes in the pathogenesis of pelvic organ prolapse and have raised the attention upon oxidative stress as an important element involved in its appearance. The advancements made over the years in terms of molecular biology have allowed researchers to investigate how the constituent elements of the pelvic-supportive structures react in conditions of oxidative stress. The purpose of this paper is to underline the importance of oxidative stress in the pathogenesis of pelvic organ prolapse, as well as to highlight the main oxidative stress molecular changes that appear at the level of the pelvic-supportive structures. Sustained mechanical stress is proven to be a key factor in the appearance of pelvic organ prolapse, correlating with increased levels of free radicals production and mitochondrial-induced fibroblasts apoptosis, the rate of cellular apoptosis depending on the intensity of the mechanical stress, and the period of time the mechanical stress is applied. Oxidative stress hinders normal cellular signaling pathways, as well as different important cellular components like proteins, lipids, and cellular DNA, therefore significantly interfering with the process of collagen and elastin synthesis.

Topics: Animals; Biomarkers; Citric Acid Cycle; Collagen; Disease Susceptibility; Elastin; Female; Humans; Mitochondria; Oxidative Stress; Pelvic Organ Prolapse

The Utah Project on Exfoliation Syndrome (UPEXS) study was created to investigate the association between exfoliation syndrome (XFS) and systemic disorders or pathologies. The study utilizes the resources of the Utah Population Database, which is linked to the Utah genealogy, a compilation of large pedigrees extending back 3 to ≥11 generations, representing most families in the state. These family members medical and health records are linked to vital records and can be used effectively to identify familial clustering of disorders, like XFS, with comorbid diseases or health-related data. There is growing evidence that XFS patients have an increased risk for systemic disorders that may reflect the systemic tissue involvement of this disease. Epidemiologic studies of individuals with XFS have reported an increased risk of various pathologies that have abnormalities in extracellular matrix metabolism and repair. For this reason, the UPEXS has focused on disorders that involve the extracellular matrix in general and elastin specifically, such as pelvic organ prolapse, atrial fibrillation, inguinal hernias, and chronic obstructive pulmonary disease. In this paper we present our results from the analysis of pelvic organ prolapse, as well as, preliminary data for atrial fibrillation.

Topics: Atrial Fibrillation; Databases, Factual; Elastin; Exfoliation Syndrome; Extracellular Matrix; Humans; Intraocular Pressure; Pelvic Organ Prolapse; Utah

To review the scientific and clinical literature to assess the basis for the use of biomaterials in stress urinary incontinence (SUI) and pelvic organ prolapse (POP). Pelvic floor diseases (PFDS), such as SUI and POP, are common and vexing disorders. While synthetic mesh-based repairs have long been considered an option for PFD treatment, and their efficacy established in randomised clinical trials, safety of its use has recently been called into question.. Using the PubMed, MEDLINE and Medical Subject Headings (MeSH) databases, we performed a critical review of English-language publications that contained the following keywords: 'pelvic organ prolapse', 'stress urinary incontinence', 'mesh', 'biomaterial', 'collagen', 'elastin' and 'extracellular matrix'. After reviewing for relevance for mesh use in the pelvis by two independent reviewers with a third available in the case of disagreement, a total of 60 articles were included in the present review.. We found that many of the potential causes of PFDs are due to altered metabolism of patient extracellular matrix (specifically collagen, elastin, and their respective enzymes) and as such, repairs using native tissue may suffer from the same abnormalities leading to a subsequent lack of repair integrity. However, mesh use is not without its unique risks. Several publications have suggested that biomaterials may undergo alteration after implantation, but these findings have not been demonstrated in the normal milieu.. While the decision for the use of synthetic mesh is scientifically sound, its benefits and risks must be discussed with the patient in an informed decision-making process.

Topics: Biocompatible Materials; Collagen; Elastin; Female; Humans; Pelvic Organ Prolapse; Randomized Controlled Trials as Topic; Surgical Mesh; Urinary Incontinence, Stress

The pathophysiology of pelvic organ prolapse (POP) is incompletely understood. The purpose of this study is to describe the current knowledge about histology of the vaginal wall and its possible involvement in the pathogenesis of pelvic organ prolapse.. Eligible studies were selected through a MEDLINE search covering January 1986 to December 2012. The research was limited to English-language publications.. Investigations of changes in the vaginal tissue that occur in women with genital prolapse are currently still limited and produced contrary results. The heterogeneity of the patients and the control groups in terms of age, parity and hormonal status, of the localization of biopsies and the histological methods as well as the lack of validation of the quantification procedures do not allow clear and definitive conclusions to be drawn.. This review shows that current knowledge of the histological changes observed in women with POP are inconclusive and relatively limited. More studies are needed in this specific field to better understand the mechanisms that lead to POP.

Topics: Collagen; Connective Tissue; Elastin; Female; Humans; Pelvic Organ Prolapse; Vagina

Pelvic floor disorders (PFDs) such as stress urinary incontinence (SUI) and pelvic organ prolapse (POP) may share a common pathophysiological process related to pelvic floor tissue laxity and loss of support. We reviewed recent literature on observed biochemical changes in women with SUI and POP, linking them to genetic predisposition. We found that studies of pelvic tissues showed differences between control subjects and women with POP and SUI in collagen and elastin structure at a molecular and fibrillar level. Studies were heterogeneous but showed a trend towards decreased collagen and elastin content. The contribution of matrix metalloproteinases to increased collagenolysis can be related to genetic polymorphisms present in higher frequency in women with PFD. Extracellular matrix (ECM) protein turnover plays a role in the development of POP and SUI, but much remains to be understood of this complex dynamic interplay of enzymes, proteins and molecules. Genotyping of candidate genes participating in ECM formation will elucidate the missing link between the manifestation of the disease and the biochemical changes observed systematically, in addition to those in the pelvic floor.

Topics: Collagen; Elastin; Extracellular Matrix; Female; Genetic Predisposition to Disease; Humans; Matrix Metalloproteinases; Pelvic Floor; Pelvic Organ Prolapse; Risk Factors; Urinary Incontinence, Stress

Pelvic organ prolapse (POP) is the descent of the bladder, uterus, and/or rectum into the vagina. POP is associated with altered vaginal fibroblast functionality and connective tissue composition in the vaginal wall. The results of surgical intervention are poor, which may be related to the lack of true restoration of the connective tissue. An innovative treatment addresses tissue repair after surgery by the introduction of a bioactive supplement that enhances the healing process through collagen and elastin deposition. As a novel strategy, we first studied the effects in an in vitro model. Here, we investigate how the presence of cell binding GRGDS (RGD) peptides on the highly biomimetic polyisocyanide (PIC) gel facilitates and promotes the function of primary vaginal fibroblasts isolated from a POP patient. Fibroblast function was analyzed in terms of morphology, proliferation, and extracellular matrix (ECM) deposition and remodeling. RGD modification of the gel facilitated cell spread and proliferation. Quantitative outcomes of the ECM content indicated increased production of collagen and elastin by fibroblasts on gels with the highest RGD density. The in vitro results suggest that PIC-RGD hydrogel application may translate into improved connective tissue healing in the pelvic floor, which is essential for its use as a regeneration promoting additive in surgery.

Topics: Collagen; Elastin; Female; Fibroblasts; Humans; Hydrogels; Pelvic Organ Prolapse; Peptides; Vagina

Women with pelvic organ prolapse (POP) have bothersome complaints that significantly affect their quality of life. While native tissue repair is associated with high recurrence rates, polypropylene knitted implants have caused specific implant-related adverse events that have detrimental, often irreversible, effects. We hypothesize that surgical outcome can be improved with a tissue-engineered solution using an absorbable implant that mimics the natural extracellular matrix (ECM) structure, releases estrogen, and activates collagen metabolism by fibroblasts as the main regulators of wound healing. To this aim, we produced electrospun poly-4-hydroxybutyrate (P4HB) scaffolds and biofunctionalized them with estradiol (E2). The cell-implant interactions relevant for POP repair were assessed by seeding primary POP vaginal fibroblasts isolated from patients on electrospun P4HB scaffolds with 1%, 2%, or 5% E2 and without E2. To test our hypothesis on whether ECM mimicking structures should improve regeneration, electrospun P4HB was compared to knitted P4HB implants. We evaluated vaginal fibroblast proliferation, ECM deposition, and metabolism by quantification of collagen, elastin, and matrix metalloproteinases and by gene expression analysis for 28 days. We established effective E2 drug loading with a steady release over time. Significantly higher cell proliferation, collagen-, and elastin deposition were observed on electrospun P4HB scaffolds as compared to knitted P4HB. For this study, physical properties of the scaffolds were more determinant on the cell response than the release of E2. These results indicate that making these electrospun P4HB scaffolds E2-releasing appears to be technically feasible. In addition, electrospun P4HB scaffolds promote the cellular response of vaginal fibroblasts and further studies are merited to assess if their use results in improved surgical outcomes in case of POP repair.

Topics: Collagen; Elastin; Female; Humans; Hydroxybutyrates; Pelvic Organ Prolapse; Quality of Life

Nondegradable transvaginal polypropylene meshes for treating pelvic organ prolapse (POP) are now generally unavailable or banned due to serious adverse events. New tissue engineering approaches combine degradable scaffolds with mesenchymal stem/stromal cells from human endometrium (eMSC). In this study, we investigate effect of microRNA-138 (miR-138) regulation on bone marrow-derived mesenchymal stem cells (BMSCs) and the efficacy of BMSC transplantation therapy in a rat POP model. We first identified FBLN5 as a target of miR-138. miR-138, fibulin-5 (FBLN5), interleukin-1β (IL-1β), and elastin expression in uterosacral ligament of POP patients and controls were detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blot analysis. After isolation and identification, BMSCs were treated to alter their expression of miR-138 or FBLN5. Proliferation of BMSCs was analyzed by CCK-8. After establishing the rat pelvic floor dysfunction (PFD) model, we evaluated efficacy of BMSC injection by applying leak point pressure (LPP) and the conscious cystometry (CMG) tests. miR-138 inhibition resulted in increased viability of BMSCs and elevated their secretion of elastin, while downregulating IL-1β expression. BMSCs with inhibited miR-138 improved LPP and conscious CMG results

Topics: Aged; Animals; Bone Marrow Cells; Cell Differentiation; Elastin; Extracellular Matrix Proteins; Female; Gene Silencing; Humans; Interleukin-1beta; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; MicroRNAs; Middle Aged; Pelvic Organ Prolapse; Rats; Rats, Sprague-Dawley; Recombinant Proteins; Signal Transduction

Current surgical therapies for pelvic organ prolapse (POP) do not repair weak vaginal tissue and just provide support; these therapies may trigger severe complications. Stem cell-based regenerative therapy, due to its ability to reconstruct damaged tissue, may be a promising therapeutic strategy for POP. The objective of this study is to evaluate whether mesenchymal stem cell (MSC) therapy can repair weak vaginal tissue in an ovariectomized rhesus macaque model.. A bilateral ovariectomy model was established in rhesus macaques to induce menopause-related vaginal injury. Ten bilaterally ovariectomized rhesus macaques were divided into two groups (n=5/group): the saline group and the MSC group. Three months after ovariectomy, saline or MSCs were injected in situ into the injured vaginal wall. The vaginal tissue was harvested 12 weeks after injection for histological and biochemical analyses to evaluate changes of extracellular matrix, microvascular density, and smooth muscle in the vaginal tissue. Biomechanical properties of the vaginal tissue were assessed by uniaxial tensile testing. Data analysis was performed with unpaired Student's t test or Mann-Whitney.. Twelve weeks after MSC transplantation, histological and biochemical analyses revealed that the content of collagen I, elastin, and microvascular density in the lamina propria of the vagina increased significantly in the MSC group compared with the saline group. And the fraction of smooth muscle in the muscularis of vagina increased significantly in the MSC group. In addition, MSC transplantation improved the biomechanical properties of the vagina by enhancing the elastic modulus.. Vaginal MSC transplantation could repair the weak vaginal tissue by promoting extracellular matrix ingrowth, neovascularization, and smooth muscle formation and improve the biomechanical properties of the vagina, providing a new prospective treatment for POP.

Topics: Animals; Elastin; Female; Humans; Macaca mulatta; Mesenchymal Stem Cell Transplantation; Pelvic Organ Prolapse; Prospective Studies

Pelvic organ prolapse (POP) decreases quality of life for many women, but its pathophysiology is poorly understood. We have previously shown that Lysyl oxidase-like 1 knockout (Loxl1 KO) mice reliably prolapse with age and increased parity, similar to women. Both this model and clinical studies also indicate that altered elastin metabolism in pelvic floor tissues plays a role in POP manifestation, although it is unknown if this is a cause or effect. Using Loxl1 KO mice, we investigated the effects of genetic absence of Loxl1, vaginal parity, and presence of POP on the expression of genes and proteins key to the production and regulation of elastic matrix. Cultured cells isolated from vaginal explants of mice were assayed with Fastin for elastic matrix, as well as RT-PCR and Western blot for expression of genes and proteins important for elastin homeostasis. Elastin synthesis significantly decreased with absence of LOXL1 and increased with parity (p < .001), but not with POP. Cells from prolapsed mice expressed significantly decreased MMP-2 (p < .05) and increased TIMP-4 (p < .05). The results suggest changes to elastin structure rather than amounts in prolapsed mice as well as poor postpartum elastin turnover, resulting in accumulation of damaged elastic fibers leading to abnormal tropoelastin deposition. POP may thus, be the result of an inability to initiate the molecular mechanisms necessary to clear and replace damaged elastic matrix in pelvic floor tissues after vaginal birth.

Topics: Amino Acid Oxidoreductases; Animals; Cells, Cultured; Disease Models, Animal; Elastin; Female; Homeostasis; Mice, Inbred C57BL; Mice, Knockout; Pelvic Organ Prolapse; Vagina

Fibroblast to myofibroblast differentiation is a key feature of wound-healing in soft tissues, including the vagina. Vaginal fibroblasts maintain the integrity of the vaginal wall tissues, essential to keep pelvic organs in place and avoid pelvic organ prolapse (POP). The micro-environment of vaginal tissues in POP patients is stiffer and has different extracellular matrix (ECM) composition than healthy vaginal tissues. In this study, we employed a series of matrices with known stiffnesses, as well as vaginal ECMs, in combination with vaginal fibroblasts from POP and healthy tissues to investigate how matrix stiffness and composition regulate myofibroblast differentiation in vaginal fibroblasts. Stiffness was positively correlated to production of α-smooth muscle actin (α-SMA). Vaginal ECMs induced myofibroblast differentiation as both α-SMA and collagen gene expressions were increased. This differentiation was more pronounced in cells seeded on POP-ECMs that were stiffer than those derived from healthy tissues and had higher collagen and elastin protein content. We showed that stiffness and ECM content regulate vaginal myofibroblast differentiation. We provide preliminary evidence that vaginal fibroblasts might recognize POP-ECMs as scar tissues that need to be remodeled. This is fundamentally important for tissue repair, and provides a rational basis for POP disease modelling and therapeutic innovations in vaginal reconstruction.

Topics: Actins; Cell Differentiation; Cells, Cultured; Collagen; Elastin; Extracellular Matrix; Female; Fibroblasts; Gene Expression; Humans; Myofibroblasts; Pelvic Organ Prolapse; Vagina

Polypropylene meshes cause severe complications in some patients. Materials that are biomechanically compatible and can better integrate into host tissues are urgently needed.. To design an oestradiol-releasing electrospun poly-l-lactic acid (PLA) mesh and evaluate its ability to stimulate new extracellular matrix and new blood vessel formation.. Human adipose derived mesenchymal cells (ADMSC) were isolated from fat. PLA meshes with micro- to nano-sized fibres containing 1%, 5%, and 10% oestradiol were constructed and used for in vitro and in vivo experiments.. The angiogenic potential of the fibrous meshes was evaluated using an in vivo chorioallantoic membrane and an in vitro chick aortic arch assays.. Oestradiol release was measured fluorometrically. The effect of fibrous meshes on proliferation and extracellular matrix (ECM) production of ADMSC was assessed using immunohistology. Mechanical properties were tested using a tensiometer.. The ultrastructure of the mesh was not affected by the inclusion of oestradiol and mechanical properties were only slightly modified. Oestradiol was released from PLA meshes over a 5-mo period. ADMSCs cultured on oestradiol-releasing PLA meshes produced more ECM involving collagen I, collagen III, and elastin. Oestradiol-releasing meshes doubled new blood vessel formation in the chorioallantoic membrane assay (p=0.001) and outgrowth of pro-angiogenic cells in the aortic arch assay (p=0.001). Further studies in longer-term animal models are required to confirm these results.. Oestradiol-releasing PLA meshes increase ECM production and stimulate angiogenesis. As such, they are promising candidate materials to be used in pelvic floor repair and to improve the initial healing phase of a repair material following implantation.. In this study, we designed a tissue engineered material to be used to support weakened pelvic floor tissues in women to avoid the complications associated with current surgical mesh. Our results showed that this material can stimulate new blood vessel formation in simple chick assays and tissue production in vitro. Both properties should help with the integration of this material into patients' tissues and merit further study in physiologically relevant animal models.

Topics: Biocompatible Materials; Chorioallantoic Membrane; Collagen; Elastin; Estradiol; Extracellular Matrix; Female; Humans; Microscopy, Electron, Scanning; Neovascularization, Physiologic; Pelvic Floor; Pelvic Organ Prolapse; Polypropylenes; Surgical Mesh; Tissue Engineering; Treatment Outcome; Urinary Incontinence, Stress

Pelvic organ prolapse (POP) is a multifactorial disorder that impairs the quality of life (QoL) of older women in particular. The purpose of this study was to elucidate the pathogenesis of POP by focusing on the extracellular matrix (ECM).. Patients were classified into two groups-with or without cervical elongation-using the POP quantification system. Specimens were obtained from 29 women with POP during hysterectomy. The expression of fibulin-5, elastin, integrin β1 (ITGβ1), lysyl oxidase-like protein-1 (LOXL1) and collagen in the vagina, uterosacral ligament, and uterine cervix was investigated by quantitative real-time polymerase chain reaction (RT-PCR) and correlation between gene levels and severity of POP examined. The location of proteins was analyzed using immunohistochemical staining and expression of fibulin-5 protein analyzed by Western blotting.. Fibulin-5 and elastin were mainly expressed in lamina propria and fibromuscular layers of the vagina and uterosacral ligament. Gene levels of fibulin-5 and ITGβ1 in uterosacral ligaments increased with severity of POP in women with cervical elongation, while no correlation was observed in women with a normal cervix. In women with uterine cervical elongation, each ECM-related gene significantly increased with POP staging. Furthermore, fibulin-5 protein also increased in the uterosacral ligament and uterine cervix.. The severity of POP and gene expression of ECM-related proteins were inversely correlated in vaginal tissue in a normal and elongated cervix. These results suggested that the differing progression of the two types of POP have a relationship with ECM-related protein.

Topics: Aged; Amino Acid Oxidoreductases; Cervix Uteri; Collagen; Elastin; Extracellular Matrix Proteins; Female; Humans; Integrin beta1; Ligaments; Middle Aged; Pelvic Organ Prolapse; RNA; Severity of Illness Index; Transcriptome; Vagina

The perineal body connects muscles from the pelvic floor and is critical for support of the lower part of the vagina and proper function of the anal canal. We determined mechanical parameters and volume fractions of main components of the human female postmenopausal perineal body.. The specimens were taken from 15 fresh female cadavers (age 74 ± 10, mean ± standard deviation). Seventy-five specimens from five regions of the perineal body were processed histologically to assess volume fractions of tissue components using stereological point testing grid. Fifteen specimens taken from the midline region were loaded uniaxially with 6 mm/min velocity until tissue rupture to determine Young's modulus of elasticity, ultimate stresses, and strains.. The perineal body was composed of collagen (29%), adipose cells (27%), elastin (7%), smooth muscle (11%), and skeletal muscle (3%). The residual tissue (19%) constituted mostly peripheral nerves, lumina of blood vessels, fibroblasts, and fibrocytes. Young's modulus of elasticity at midline region was 18 kPa (median) at small and 232 kPa at large deformations, respectively. The ultimate stress was 172 kPa and the ultimate strain was 1.4.. We determined the structural and mechanical parameters of the perineal body. The resultant data could be used as input for models simulating pelvic floor prolapse or dysfunction.

Topics: Adipose Tissue; Aged; Aged, 80 and over; Anal Canal; Biomechanical Phenomena; Cadaver; Collagen; Elasticity; Elastin; Female; Humans; Middle Aged; Muscle, Skeletal; Muscle, Smooth; Pelvic Floor; Pelvic Organ Prolapse; Perineum; Postmenopause; Vagina

Pelvic organ prolapse, various types of incontinence (urinary incontinence, defecatory dysfunction), chronic cystourethritis, and sexual dysfunctions remain between the most common disorders in urogynecology. Currently, it is believed that the nature and number of births plays a major role in their development. Moreover, after these events, pelvic floor tissues may not recover to their original statuses. The close anatomical relationship among the vaginal wall, bladder and rectum often contribute to the emergence of anatomical-functional failure of adjacent organs and systems.. The aim of this study was to investigate the effect of consecutive pregnancies on pelvic floor soft tissues, conducting biomechanical and histological analysis. Fifteen Swifter ewes: virgins, parous and pregnant were used. Samples, for uniaxial tension tests and histological analysis, were cut out from fresh tissue. A description of the mechanical properties of native tissue was obtained from the stress-strain curve. Histological samples were stained with Miller's Elastica staining and analyzed using ImageJ software. Collagen, elastin, and smooth muscle contents (%) were analyzed along the full wall thickness of the selected organs. The links between mechanical properties of the soft tissues and histological parameters were analyzed.. Mechanically, vaginal wall tissue and cervix of pregnant sheep were more compliant. In contrast, bladder and rectum became stiffer and had the highest total collagen content. Parous sheep rectum and bladder were stiffer, compared to virgin sheep.. Tensile strength appears to be linked to total collagen content. Elastin and smooth muscle show a direct influence on tissue compliance.

Topics: Animals; Biomechanical Phenomena; Cervix Uteri; Collagen; Elasticity; Elastin; Female; Pelvic Organ Prolapse; Pelvis; Pregnancy; Pregnancy, Animal; Rectum; Sheep, Domestic; Tensile Strength; Urinary Bladder; Vagina

Synthetic non-absorbable meshes are widely used to augment surgical repair of pelvic organ prolapse (POP) and stress urinary incontinence (SUI), but these meshes are associated with serious complications. This study compares the attachment and extracellular matrix (ECM) production of adipose-derived stem cells (ADSCs) on different biodegradable nanomaterials to develop tissue engineered repair materials.. Rat ADSCs were isolated and cultured on electrospun poly-L-lactic acid (PLA) and electrospun poly(L-lactide)-trimethylene carbonate-gycolide (PLTG) terpolymers for 1 and 2 weeks. Samples were tested for cell proliferation (cell counting kit-8), microstructure, and morphology (scanning electron microscopy), production of ECM components (immunostaining for collagen I, collagen III, and elastin) and biomechanical properties (uniaxial tensile methods).. The ADSCs showed good attachment and proliferation on both PLA and PLTG scaffolds. The production of collagen I and collagen III on both scaffolds was greater at 14 days than at 7 days and was greater on PLTG scaffolds than on PLA scaffolds, but these differences were not significant. The addition of ADSCs onto scaffolds led to a significant increase in the biomechanical properties of both PLA and PLTG scaffolds compared with unseeded scaffolds.. These data support the use of both PLA and PLTG as tissue-engineered repair materials for POP or SUI.

Topics: Adipose Tissue; Animals; Cell Proliferation; Collagen Type I; Collagen Type II; Disease Models, Animal; Elastin; Female; Humans; Pelvic Organ Prolapse; Rats; Rats, Sprague-Dawley; Stem Cells; Tissue Engineering; Tissue Scaffolds; Triglycerides; Urinary Incontinence

Pelvic organ prolapse (POP) is a global health problem that may seriously impact the quality of life of the sufferer. The present study aimed to investigate the potential mechanisms underlying alterations in extracellular matrix (ECM) metabolism in the pathogenesis of POP, by investigating the expression of ECM components in human parametrial ligament fibroblasts (hPLFs) subject to various mechanical strain loads. Fibroblasts derived from parametrial ligaments were cultured from patients with POP and without malignant tumors, who underwent vaginal hysterectomy surgery. Fibroblasts at generations 3‑6 of exponential phase cells were selected, and a four‑point bending device was used for 0, 1,333 or 5,333 µ mechanical loading of cells at 0.5 Hz for 4 h. mRNA and protein expression levels of collagen type I α 1 chain (COL1A1), collagen type III α 1 chain (COL3A1), elastin, matrix metalloproteinase (MMP) ‑2 and ‑9, and transforming growth factor (TGF)‑β1 were detected by reverse transcription‑quantitative polymerase chain reaction and western blotting, respectively. Under increased mechanical strain (5,333 µ), mRNA and protein expression levels of COL1A1, COL3A1 elastin and TGF‑β1 decreased, particularly COL1A1; however, mRNA and protein expression levels of MMP‑2 and ‑9 were significantly increased, compared with the control group (0 µ strain). Following 1,333 µ mechanical strain, mRNA and protein expression levels of COL1A1, COL3A1 elastin and MMP‑2 increased, and MMP‑9 decreased, whereas no significant differences were observed in TGF‑β1 mRNA and protein expression levels. In conclusion, ECM alterations may be involved in pathogenesis of POP, with decreased synthesis and increased degradation of collagen and elastin. Furthermore, the TGF‑β1 signaling pathway may serve an important role in this process and thus may supply a new target and strategy for understanding the etiology and therapy of POP.

Topics: Cells, Cultured; Collagen Type I; Collagen Type I, alpha 1 Chain; Collagen Type III; Elastin; Extracellular Matrix; Fibroblasts; Humans; Ligaments; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Neoplasms; Pelvic Organ Prolapse; Stress, Mechanical; Transforming Growth Factor beta1

The present study aimed to reveal the metabolic alterations of the extracellular matrix (ECM) in uterosacral ligament (USL) with pelvic organ prolapse (POP) and to explore the role of transforming growth factor‑β1 (TGF‑β1) in pathogenesis of POP. For this purpse, 60 participants who underwent hysterectomy for benign indications were enrolled, 30 of which had symptomatic POP (grade II, III or IV) and composed the POP group, and the other 30 had asymptomatic POP (grade I or less) and served as the controls. Collagen fibers, elastin,matrix metalloproteinase (MMP)‑2/9, tissue inhibitor of matrix metalloproteinases (TIMP)‑2 and TGF‑β1 were examined by Masson's trichrome staining, immunohistochemistry and RT-qPCR using USL biopsies. In vitro, human USL fibroblasts (hUSLFs) were primary cultured, pre-treated with recombinant TGF‑β1 (0, 5, or 10 ng/ml) and then subjected to cyclic mechanical stretching (CMS; 0 or 5,333 µε strain). Changes in the expression levels of collagen type I/III, elastin, TIMP‑2, MMP‑2/9 and Smad were detected. Our results revealed that at the tissue level, the expression of collagen fibers, elastin, TIMP‑2 and TGF‑β1 was significantly reduced in the POP group, while the activities of MMP‑2/9 were significantly upregulated, compared with the control group. Statistical analysis indicated that the mRNA expression of TGF‑β1 inversely correlated with the severity of POP partially. Our in vitro experimental data demonstrated that a CMS of 5333 µε strain promoted the degradation of ECM proteins, inhibited the synthesis of TIMP‑2, and upregulated the proteolytic activities of MMP‑2/9. Pre-treatment with TGF‑β1 attenuated the loss of ECM by stimulating the synthesis of TIMP‑2 and inhibiting the activities of MMP‑2/9 through the TGF‑β1/Smad3 signaling pathway. On the whole, our data indicate that the reduced anabolism and increased catabolism of ECM proteins in USL are the pathological characteristics of POP. TGF‑β1 not only has a specific value in predicting the severity of POP, but should also be considered as a novel therapeutic target for POP.

Topics: Cells, Cultured; Collagen; Elastin; Extracellular Matrix; Female; Humans; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Middle Aged; Pelvic Organ Prolapse; Proteolysis; Signal Transduction; Smad Proteins; Tissue Inhibitor of Metalloproteinase-2; Transforming Growth Factor beta1

The aim of the study was to correlate histological and biomechanical characteristics of the vaginal wall in women with pelvic organ prolapse (POP).. Tissue samples were collected from the anterior [point Ba; POP Questionnaire (POP-Q)] and/or posterior (point Bp; POP-Q) vaginal wall of 15 women who underwent vaginal surgery for POP. Both histological and biomechanical assessments were performed from the same tissue samples in 14 of 15 patients. For histological assessment, the density of collagen and elastin fibers was determined by combining high-resolution virtual imaging and computer-assisted digital image analysis. For biomechanical testing, uniaxial tension tests were performed to evaluate vaginal tissue stiffness at low (C0) and high (C1) deformation rates.. Biomechanical testing highlights the hyperelastic behavior of the vaginal wall. At low strains (C0), vaginal tissue appeared stiffer when elastin density was low. We found a statistically significant inverse relationship between C0 and the elastin/collagen ratio (p = 0.048) in the lamina propria. However, at large strain levels (C1), no clear relationship was observed between elastin density or elastin/collagen ratio and stiffness, likely reflecting the large dispersion of the mechanical behavior of the tissue samples.. Histological and biomechanical properties of the vaginal wall vary from patient to patient. This study suggests that elastin density deserves consideration as a relevant factor of vaginal stiffness in women with POP.

Topics: Aged; Biomechanical Phenomena; Collagen; Elasticity; Elastin; Female; Humans; Image Processing, Computer-Assisted; Middle Aged; Pelvic Organ Prolapse; Stress, Mechanical; Vagina

Pelvic floor dysfunction (PFD) is a group of clinical conditions including stress urinary incontinence (SUI) and pelvic organ prolapse (POP). The abnormality of collagen and elastin metabolism in pelvic connective tissues is implicated in SUI and POP.. To reconstitute the connective tissues with normal distribution of collagen and elastin, we transduced elastin to bone marrow-derived mesenchymal stem cells (BMSC). Elastin-expressing BMSCs were then differentiated to fibroblasts using bFGF, which produced collagen and elastin. To achieve the sustained release of bFGF, we formulated bFGF in poly (lactic-co-glycolic acid) (PLGA) nanoparticles (NP).. In an in vitro cell culture system of 7 days, when no additional bFGF was administrated, the initial PLGA-loaded bFGF NP induced prolonged production of collagen and elastin from elastin-expressing BMSCs. In vivo, co-injection of PLGA-loaded bFGF NP and elastin-expressing BMSCs into the PFD rats significantly improved the outcome of urodynamic tests. Together, these results provided an efficient model of connective tissue engineering using BMSC and injectable PLGA-loaded growth factors.. Our results provided the first instance of a multidisciplinary approach, combining both stem cell and nanoparticle technologies, for the treatment of PFD.

Topics: Animals; Bone Marrow Cells; Cell Differentiation; Collagen; Disease Models, Animal; Drug Compounding; Elastin; Female; Fibroblast Growth Factor 2; Fibroblasts; Gene Expression; Lactic Acid; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Nanoparticles; Pelvic Organ Prolapse; Polyglycolic Acid; Polylactic Acid-Polyglycolic Acid Copolymer; Rats; Rats, Sprague-Dawley; Transduction, Genetic; Urinary Incontinence

Mesh-augmented vaginal surgery for treatment of pelvic organ prolapse (POP) does not meet patients' needs. This study aims to test the hypothesis that fascia tissue engineering using adipose-derived stem cells (ADSCs) might be a potential therapeutic strategy for reconstructing the pelvic floor.. Human ADSCs were isolated, differentiated, and characterized in vitro. Both ADSCs and fibroblastic-differentiated ADSCs were used to fabricate tissue-engineered fascia equivalents, which were then transplanted under the back skin of experimental nude mice.. ADSCs prepared in our laboratory were characterized as a group of mesenchymal stem cells. In vitro fibroblastic differentiation of ADSCs showed significantly increased gene expression of cellular collagen type I and elastin (p < 0.05) concomitantly with morphological changes. By contrast, ADSCs cultured in control medium did not demonstrate these changes. Both of the engrafted fascia equivalents could be traced up to 12 weeks after transplantation in the subsequent animal study. Furthermore, the histological outcomes differed with a thin (111.0 ± 19.8 μm) lamellar connective tissue or a thick (414.3 ± 114.9 μm) adhesive fibrous tissue formation between the transplantation of ADSCs and fibroblastic-differentiated ADSCs, respectively. Nonetheless, the implantation of a scaffold without cell seeding (the control group) resulted in a thin (102.0 ± 17.1 μm) fibrotic band and tissue contracture.. Our results suggest the ADSC-seeded implant is better than the implant alone in enhancing tissue regeneration after transplantation. ADSCs with or without fibroblastic differentiation might have a potential but different role in fascia tissue engineering to repair POP in the future.

Topics: Adipose Tissue; Animals; Cell Differentiation; Collagen Type I; Elastin; Fascia; Female; Fibroblasts; Guided Tissue Regeneration; Humans; Mice; Mice, Nude; Pelvic Organ Prolapse; Stem Cells; Tissue Engineering; Tissue Scaffolds; Tissue Transplantation

Synthetic non-absorbable meshes are widely used to augment surgical repair of stress urinary incontinence (SUI) and pelvic organ prolapse (POP); however, there is growing concern such meshes are associated with serious complications. This study compares the potential of two autologous cell sources for attachment and extra-cellular matrix (ECM) production on a biodegradable scaffold to develop tissue engineered repair material (TERM).. Human oral fibroblasts (OF) and human adipose-derived stem cells (ADSC) were isolated and cultured on thermo-annealed poly-L-lactic acid (PLA) scaffolds for two weeks under either unrestrained conditions or restrained (either with or without intermittent stress) conditions. Samples were tested for cell metabolic activity (AlamarBlue® assay), contraction (serial photographs analyzed with image J software), total collagen production (Sirius red assay), and production of ECM components (immunostaining for collagen I, III, and elastin; and scanning electron microscopy) and biomechanical properties (BOSE tensiometer). Differences were statistically tested using two sample t-test.. Both cells showed good attachment and proliferation on scaffolds. Unrestrained scaffolds with ADSC produced more total collagen and a denser homogenous ECM than OF under same conditions. Restrained conditions (both with and without intermittent stress) gave similar total collagen production, but improved elastin production for both cells, particularly OF. The addition of any cell onto scaffolds led to an increase in biomechanical properties of scaffolds compared to unseeded scaffolds.. OF and ADSC both appear to be suitable cell types to combine with biodegradable scaffolds, in the development of a TERM for the treatment of SUI and POP. Neurourol. Urodynam. 33:531-537, 2014. © 2013 Wiley Periodicals, Inc.

Topics: Absorbable Implants; Cell Proliferation; Collagen Type I; Collagen Type II; Collagen Type III; Elastin; Extracellular Matrix; Fibroblasts; Humans; Lactic Acid; Microscopy, Electron, Scanning; Mouth; Pelvic Organ Prolapse; Polyesters; Polymers; Stem Cells; Subcutaneous Fat; Tissue Engineering; Tissue Scaffolds; Urinary Incontinence, Stress

Aim of the study was the comparison of the connective tissue components in the uterine ligaments between the women with and without pelvic organ prolapse.. This study included 15 postmenopausal women without pelvic organ prolapse who underwent total abdominal hysterectomy and bilateral salpingo-oophorectomy for benign pathologies (control group) and 15 postmenopausal women who underwent vaginal hysterectomy because of pelvic organ prolapse (POP group). During the operation, samples for histological examination were taken from the cardinal and sacrouterine ligaments. The contents of collagen type I, collagen type III and elastin, and number and mitotic activity of fibroblasts in uterine ligaments were histopatholocially determined.. The collagen type I content for the POP group was significantly higher than that of the control group. Collagen type III content, elastin content, and number and mitotic activity of fibroblasts were not significantly different between the control and POP groups.. Collagen type I was found to be increased in the uterine ligaments of the POP group. The other connective tissue components did not differ between the two groups.

Topics: Aged; Case-Control Studies; Collagen Type I; Collagen Type III; Connective Tissue; Elastin; Female; Fibroblasts; Humans; Hysterectomy; Ligaments; Middle Aged; Mitosis; Pelvic Organ Prolapse; Postmenopause; Prospective Studies; Uterus

The purpose of this study was to analyze the histomorphometric properties of the vaginal wall in women with pelvic organ prolapse (POP).. In 15 women undergoing surgery for POP, full-thickness biopsies were collected at two different sites of location from the anterior and/or posterior vaginal wall. Properties of the precervical area (POP-Q point C/D) were compared with the most distal portion of the vaginal wall (POP-Q point Ba/Bp) using histological staining and immunohistochemistry. The densities of total collagen fibers, elastic fibers, smooth muscle cells, and blood vessels were determined by combining high-resolution virtual imaging and computer-assisted digital image analysis.. The mean elastin density was significantly decreased in the lamina propria and muscularis layer of the vaginal wall from the most distal portion of the prolapsed vaginal wall compared with the precervical area. This difference was statistically significant in the lamina propria for both anterior (8.4 ± 1.2 and 12.1 ± 2.0, p = 0.048) and posterior (6.8 ± 0.5 and 10.1 ± 1.4, p = 0.040) locations, and in the muscularis for the anterior (5.2 ± 0.4 and 8.4 ± 1.2, p = 0.009) vaginal wall. There were no statistically significant differences in the mean densities of collagen fibers, smooth muscle cells or blood vessels between the two locations.. In this study, we observed changes in elastin density in two different locations of the vaginal wall from women with POP. The histomorphometric properties of the vaginal wall can be variable from one place to another in the same patient. This result supports the existence of most vulnerable locations within the vaginal wall and the potential benefit of site-specific prolapse surgery.

Topics: Aged; Biopsy; Collagen; Elasticity; Elasticity Imaging Techniques; Elastin; Female; Humans; Image Processing, Computer-Assisted; Middle Aged; Muscle, Smooth; Pelvic Organ Prolapse; Vagina

To determine the composition of the vaginal wall and to explore the connective tissue layer, muscularis and basement membrane in relation to the pathogenesis of pelvic organ prolapse (POP).. We performed a histopathological study with respect to the composition of the vaginal wall of 33 patients and 9 controls. Tissue samples from the vaginal wall were analysed for collagens II and IV, desmin, elastin, fibronectin, smooth muscle actin (SMA) and transforming growth factor (TGF)-β1 using (immuno)histochemistry. Morphometric analyses were also performed.. Morphometric characteristics and expression of SMA, TGF-β1, elastin and collagen II were significantly altered in women with POP.. Our results suggest that there could be an altered tissue composition of the vaginal wall in women with POP. SMA expression could play a role in the pathogenesis of POP. The alterations in elastin and TGF-β1 expression are likely a result of POP.

Topics: Actins; Adult; Aged; Antigens; Case-Control Studies; Collagen Type II; Elastin; Female; Humans; Immunohistochemistry; Middle Aged; Muscle, Smooth; Pelvic Organ Prolapse; Transforming Growth Factor beta1; Vagina

Pelvic organ prolapse is associated with defects in connective tissues, including elastic fibers. The purpose of this study was to investigate the expression of fibulin-5 and lysyl oxidase-like 1 (LOXL1), which play an essential role in the synthesis and assembly of elastic fibers, in the uterosacral ligaments of women with advanced pelvic organ prolapse (POP) compared with controls.. Specimens were obtained prospectively during transvaginal or abdominal hysterectomy from 30 women with advanced pelvic organ prolapse and 30 controls matched to the POP group for age and parity among postmenopausal women with benign gynecologic diseases. The expressions of elastin, fibulin-5 and LOXL1 in uterosacral ligaments were measured by immunohistochemistry.. We detected a decreased, sometimes absent, expression of fibulin-5 and LOXL1 in the uterosacral ligaments of women with POP, despite a positive expression of elastin. There was a decrease in positive percentage of LOXL1 in the POP group (23.3%) compared with the controls (60%) (P = 0.004). With immunolabeling intensity classified as negative, weak, moderate or strong, there was a decrease in the expression of fibulin-5 in the POP group (P = 0.049). We also detected a significantly decreased expression of LOXL1 in the POP group (P = 0.001).. There was decreased expression of fibulin-5 and LOXL1 in the uterosacral ligaments of patients with pelvic organ prolapse, while the elastin expression was equivalent, which may suggest the possibility of defects in elastic fiber remodeling in the postpartum period and contribute to POP.

Topics: Aged; Amino Acid Oxidoreductases; Cohort Studies; Down-Regulation; Elastin; Extracellular Matrix Proteins; Female; Humans; Hysterectomy; Ligaments; Middle Aged; Pelvic Organ Prolapse; Postmenopause; Prospective Studies; Sacrum; Severity of Illness Index; Uterus

The aim of this study was to identify any histological changes between the prolapsed and non-prolapsed vaginal skin to enable us to answer the broader question of whether to excise or re-suture the prolapsed tissue during vaginal reconstructive surgery. Two tissue samples were taken from 20 women who underwent prolapse surgery, one prolapsed and another non-prolapsed tissue. The samples were analysed blind histologically by a pathologist. Our study showed statistically significant differences in three tissue components namely myofibroblast differentiation (0.047), elastin (0.048) and collagen (0.095), depending on whether the tissue was prolapsed or non-prolapsed. However, the study had certain limitations and practical difficulties faced in histological approach of tissue analysis. Histology alone is an insufficient indicator to answer our question and definitive understanding of the prolapsed tissue may result from a randomised study on a larger study population.

Topics: Adult; Aged; Cell Differentiation; Collagen; Elastin; Female; Humans; Middle Aged; Muscle, Smooth; Myofibroblasts; Pelvic Organ Prolapse; Plastic Surgery Procedures; Skin; Vagina

Although there are many studies about the effects of vaginal birth, the effects of menopause on pelvic floor support have not been identified. We compared elastin metabolism in the uterosacral ligament of women with and without pelvic organ prolapse, and defined the menopausal regulation of this process.. The study group consisted of 35 women who underwent hysterectomy for pelvic organ prolapse. The control group consisted of 39 women without pelvic organ prolapse. A questionnaire was administered to assess age, parity, body mass index, and menopausal status. Expression levels of mRNA, and protein for neutrophil elastase, matrix metalloproteinase-2, and matrix metalloproteinase-9 were determined by real-time quantitative polymerase chain reaction and ELISA, respectively, using uterosacral ligament samples from each patient. Expression of alpha-1-antitrypsin, an inhibitor of neutrophil elastase, was also determined. ANOVA, the Kruskal-Wallis test and multivariate linear regression were used for statistical analysis.. Expression of neutrophil elastase and matrix metalloproteinase-2 mRNA was higher in women with pelvic organ prolapse than in those without pelvic organ prolapse. Compared to before menopause, neutrophil elastase and matrix metalloproteinase-2 showed a significant decrease in postmenopausal women without pelvic organ prolapse, although these remained increased in postmenopausal women with pelvic organ prolapse. Alpha-1-antitrypsin was significantly less in postmenopausal women with pelvic organ prolapse than in postmenopausal women without pelvic organ prolapse. The activities of neutrophil elastase, matrix metalloproteinase-2 and matrix metalloproteinase-9 were increased in women with pelvic organ prolapse, and these trends were similar to neutrophil elastase and matrix metalloproteinase-2 expression even after adjustment for age, parity and menopausal status.. After menopause increased elastolytic protease has a significant role in the development of pelvic organ prolapse.

Topics: Aged; Analysis of Variance; Body Mass Index; Case-Control Studies; Elastin; Enzyme-Linked Immunosorbent Assay; Female; Humans; Hysterectomy; Leukocyte Elastase; Ligaments; Linear Models; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Parity; Pelvic Organ Prolapse; Polymerase Chain Reaction; Postmenopause; Premenopause; RNA, Messenger; Statistics, Nonparametric; Surveys and Questionnaires

Pelvic organ prolapse (POP) is a disabling disorder in women characterized by a loss of pelvic floor support leading to the herniation of the uterus into or through the vagina. POP is a complex problem that likely involves multiple mechanisms, and available therapies are limited. In this issue of the JCI, Budatha et al. explore the dual role carried out by fibulin-5 in facilitating the assembly of normal elastic fibers and inhibiting MMP-9 activity, revealing a new mechanism critical to the maintenance of pelvic organ support.

Topics: Animals; Disease Models, Animal; Elastic Tissue; Elastin; Extracellular Matrix Proteins; Female; Humans; Matrix Metalloproteinase 9; Mice; Mice, Transgenic; Pelvic Floor; Pelvic Organ Prolapse; Protein Structure, Tertiary; Tropoelastin; Vagina

Topics: Animals; Elastic Tissue; Elastin; Extracellular Matrix; Extracellular Matrix Proteins; Female; Humans; Matrix Metalloproteinase 9; Mice; Mice, Knockout; Pelvic Organ Prolapse; Recombinant Proteins; Vagina

The objective of this study is to define the diagnosis of hypertrophic cervical elongation clinically and to perform histochemical and histological evaluations of patients with and without hypertrophic cervical elongation.. This prospective study was conducted at Louisiana State University between December 2005 and May 2008. Fourteen women with cervical elongation and 28 women without prolapse were studied.. The amounts of elastin, collagen, and smooth muscle did not differ between study and control groups. Estrogen and progesterone receptor content in cervical elongation were elevated compared to the cervix of women without prolapse. Hypertrophic cervical elongation was defined as the difference between point C and point D of the Pelvic Organ Prolapse Quantification system greater than 8 cm.. Estrogen and progesterone receptor levels are greater in women with hypertrophic cervical elongation compared with a normal cervix.

Topics: Adult; Case-Control Studies; Cervix Uteri; Collagen; Elastin; Female; Humans; Hypertrophy; Middle Aged; Pelvic Organ Prolapse; Pilot Projects; Prospective Studies; Receptors, Estrogen; Receptors, Progesterone; Retrospective Studies

To investigate the expression and significance of elastin and fibulin-5 in anterior vaginal tissue of women with pelvic organ prolapse (POP).. Between November 2006 and June 2008, 68 patients with POP underwent surgical treatment in Shengjing Hospital of China Medical University were enrolled in this study, who were classified into 10 patients with grade I, 21 patients with grade II, 25 patients with grade III and 12 patients with grade IV in accordance with pelvic organ prolapse quantitation (POP-Q). Meanwhile, 18 cases with early cervical cancer at stage of I b were treated by total hysterectomy and bilateral salpingo-oophorectomy, their anterior vaginal tissues were selected as controls. Immunohistochemical staining was performed to detect the expression of elastin and fibulin-5.. (1) Elastin and fibulin-5 were mainly expressed at extracellular matrix(ECM). (2) The positive rate of fibulin-5 expression in anterior vaginal wall were 5% (2/37) in grade III/IV and 26% (8/31) in grade I/IV POP patients, which reached statistical difference (P = 0.035). However, no statistical different expression was found between postmenopausal (13%, 8/60) and non-menopausal patients (2/8), vaginal delivery < or =2 (19%, 5/27) and >2 patients (12%, 5/41, P > 0.05). (3) The positive rate of elastin expression in anterior vaginal wall in POP group was 31% (21/68), which was significantly lower than 72% (13/18) of control group (P = 0.002). Among POP group, 19% (7/37) of elastin expression in grade III/IV POP was significantly lower than 45% (14/31) in grade I/II of POP patients. However, no statistical difference was found between postmenopausal (30%, 18/60) and non-menopausal patients (3/8), vaginal delivery < or =2 (26%, 7/27) and >2 patients(34%, 14/41, P > 0.05). (4) In POP group, both positive expression of fibulin-5 and elastin of anterior vaginal wall was in 6 cases, both negative expression of fibulin-5 and elastin was in 43 cases. It was illustrated that elastin and fibulin-5 had an positive relationship (P = 0.031).. The decreased expression of elastin and fibulin-5 was correlated with degree of POP, which indicated that elastin and fibulin-5 may play a role in the pathogenesis of POP.

Topics: Aged; Carcinoma, Squamous Cell; Connective Tissue; Elastin; Extracellular Matrix Proteins; Female; Humans; Immunohistochemistry; Menopause; Middle Aged; Pelvic Organ Prolapse; Severity of Illness Index; Uterine Cervical Neoplasms; Vagina