Vaginal Distension Research Articles

The effect of tramadol on sneeze-induced urethral continence reflex through μ-opioid receptors in the spinal cord in rats.

We examined the efficacy of tramadol on the urethral reflex during sneezing, as well as the role of µ-opioid receptors in the spinal cord, in rats. Forty-one female Sprague-Dawley rats were used. The rats were divided into normal female rats and rats with vaginal distension (VD), which mimics stress urinary incontinence (SUI) in humans. Under urethane anesthesia, the sneeze-induced amplitude of urethral responses (AUR) and baseline pressure (BP) were examined after intravenous injection of tramadol using a microtransducer-tipped catheter in both rat groups. The effect of intrathecal cyprodime, a selective µ-opioid receptor antagonist, following intravenous tramadol injection was examined in normal rats. The tilt leak point pressure (tilt LPP) after intravenous tramadol injection was also evaluated in both groups. In normal rats, tramadol enhanced the AUR and BP by 33.2% and 19.5%, respectively. Tramadol also increased BP by 13.9% in rats with VD, but it did not change AUR. Intrathecal cyprodime alone did not change AUR, but it decreased BP. However, tramadol-provoked increments in AUR were blocked by intrathecal cyprodime, while BP was recovered to the level that it was before administration of cyprodime. Tramadol was associated with a significant elevation in tilt LPP: 24.8% and 19.5% in normal and VD rats, respectively. These findings suggest that tramadol effectively enhances the AUR at the spinal level and BP peripherally. Therefore, stimulation of the spinal µ-opioid receptors may be useful for the treatment of SUI.

Pelvic muscles’ mechanical response to strains in the absence and presence of pregnancy-induced adaptations in a rat model

Maternal birth trauma to the pelvic floor muscles is thought to be consequent to mechanical demands placed on these muscles during fetal delivery that exceed muscle physiological limits. The above is consistent with studies of striated limb muscles that identify hyperelongation of sarcomeres, the functional muscle units, as the primary cause of mechanical muscle injury and resultant muscle dysfunction. However, pelvic floor muscles' mechanical response to strains have not been examined at a tissue level. Furthermore, we have previously demonstrated that during pregnancy, rat pelvic floor muscles acquire structural and functional adaptations in preparation for delivery, which likely protect against mechanical muscle injury by attenuating the strain effect. We sought to determine the mechanical impact of parturition-related strains on pelvic floor muscles' microstructure, and test the hypothesis that pregnancy-induced adaptations modulate muscle response to strains associated with vaginal delivery. Three-month-old Sprague-Dawley late-pregnant (N= 20) and nonpregnant (N= 22) rats underwent vaginal distention, replicating fetal crowning, with variable distention volumes. Age-matched uninjured pregnant and nonpregnant rats served as respective controls. After sacrifice, pelvic floor muscles, which include coccygeus, iliocaudalis, and pubocaudalis, were fixed in situ and harvested for fiber and sarcomere length measurements. To ascertain the extent of physiological strains during spontaneous vaginal delivery, analogous measurements were obtained in intrapartum rats (N= 4) sacrificed during fetal delivery. Data were compared with repeated measures and 2-way analysis of variance, followed by pairwise comparisons, with significance set at P < .05. Gross anatomic changes were observed in the pelvic floor muscles following vaginal distention, particularly in the entheseal region of pubocaudalis, which appeared translucent. The above appearance resulted from dramatic stretch of the myofibers, as indicated by significantly longer fiber length compared to controls. Stretch ratios, calculated as fiber length after vaginal distention divided by baseline fiber length, increased gradually with increasing distention volume. Paralleling these macroscopic changes, vaginal distention resulted in acute and progressive increase in sarcomere length with rising distention volume. The magnitude of strain effect varied by muscle, with the greatest sarcomere elongation observed in coccygeus, followed by pubocaudalis, and a smaller increase in iliocaudalis, observed only at higher distention volumes. The average fetal rat volume approximated 3 mL. Pelvic floor muscle sarcomere lengths in pregnant animals undergoing vaginal distention with 3 mL were similar to intrapartum sarcomere lengths in all muscles (P > .4), supporting the validity of our experimental approach. Vaginal distention resulted in dramatically longer sarcomere lengths in nonpregnant compared to pregnant animals, especially in coccygeus and pubocaudalis (P < .0001), indicating significant attenuation of sarcomere elongation in the presence of pregnancy-induced adaptations in pelvic floor muscles. Delivery-related strains lead to acute sarcomere elongation, a well-established cause of mechanical injury in skeletal muscles. Sarcomere hyperelongation resultant from mechanical strains is attenuated by pregnancy-induced adaptations acquired by the pelvic floor muscles prior to parturition.

Protective role of Nrf2 against mechanical-stretch-induced apoptosis in mouse fibroblasts: a potential therapeutic target of mechanical-trauma-induced stress urinary incontinence.

We investigated the protective effect and underlying molecular mechanism of nuclear factor-E2-related factor 2 (Nrf2) against mechanical-stretch-induced apoptosis in mouse fibroblasts. Normal cells, Nrf2 silencing cells, and Nrf2 overexpressing cells were respectively divided into two groups-nonintervention and cyclic mechanical strain (CMS)-subjected to CMS of 5333μ (1.0Hz for 4h), six groups in total (control, CMS, shNfe212, shNfe212+CMS, LV-shNfe212, and LV-shNfe212+CMS). After treatment, cell apoptosis; cell-cycle distribution; expressions of Nrf2, Bax, Bcl-2, Cyt-C, caspase-3, caspase-9, cleaved-caspase-3, and cleaved-caspase-9; mitochondrial membrane potential (ΔΨm); reactive oxygen species (ROS); and malondialdehyde (MDA) levels were measured. Thirty virgin female C57BL/6 mice were divided into two groups: control (without intervention) and vaginal distension (VD) groups, which underwent VD for 1 h with an 8-mm dilator (0.3ml saline). Leak-point pressure (LPP) was tested on day 7 after VD; Nrf2 expression, apoptosis, and MDA levels were then measured in urethra and anterior vaginal wall. Mechanical stretch decreased Nrf2 messenger RNA (mRNA) and protein expressions. Overexpression of Nrf2 alleviated mechanical-stretch-induced cell apoptosis; S-phase arrest of cell cycle; up-regulation of Bax, cytochrome C (Cyt-C), ROS, MDA, ratio of cleaved-caspase-3/caspase-3 and cleaved-caspase-9/caspase-9; and exacerbated the decrease of Bcl2 and ΔΨm in L929 cells. On the contrary, silencing of Nrf2 showed opposite effects. Besides, VD reduced LPP levels and Nrf2 expression and increased cell apoptosis and MDA generation in the urethra and anterior vaginal wall. Nrf2 exhibits a protective role against mechanical-stretch -induced apoptosis on mouse fibroblasts, which might indicate a potential therapeutic target of mechanical-trauma-induced stress urinary incontinence (SUI).

Controlled release of insulin-like growth factor 1 enhances urethral sphincter function and histological structure in the treatment of female stress urinary incontinence in a rat model.

To determine the effects of controlled release of insulin-like growth factor 1 (IGF-1) from alginate-poly-L-ornithine-gelatine (A-PLO-G) microbeads on external urethral sphincter (EUS) tissue regeneration in a rat model of stress urinary incontinence (SUI), as SUI diminishes the quality of life of millions, particularly women who have delivered vaginally, which can injure the urethral sphincter. Despite several well-established treatments for SUI, growth factor therapy might provide an alternative to promote urethral sphincter repair. In all, 44 female Sprague-Dawley rats were randomised into four groups: vaginal distension (VD) followed by periurethral injection of IGF-1-A-PLO-G microbeads (VD + IGF-1 microbeads; 1 × 104 microbeads/1 mL normal saline); VD + empty microbeads; VD + saline; or sham-VD + saline (sham). Urethral function (leak-point pressure, LPP) was significantly lesser 1 week after VD + saline [mean (sem) 23.9 (1.3) cmH2 O] or VD + empty microbeads [mean (sem) 21.7 (0.8) cmH2 O) compared to the sham group [mean (sem) 44.4 (3.4) cmH2 O; P < 0.05), indicating that the microbeads themselves do not create a bulking or obstructive effect in the urethra. The LPP was significantly higher 1 week after VD + IGF-1 microbeads [mean (sem) 28.4 (1.2) cmH2 O] compared to VD + empty microbeads (P < 0.05), and was not significantly different from the LPP in sham rats, demonstrating an initiation of a reparative effect even at 1 week after VD. Histological analysis showed well-organised skeletal muscle fibres and vascular development in the EUS at 1 week after VD + IGF-1 microbeads, compared to substantial muscle fibre attenuation and disorganisation, and less vascular formation at 1 week after VD + saline or VD + empty microbeads. Periurethral administration of IGF-1-A-PLO-G microbeads facilitates recovery from SUI by promoting skeletal myogenesis and revascularisation. This therapy is promising, but detailed and longer term studies in animal models and humans are needed.

Effects of multiple simulated birth traumas on urethral continence function in rats.

Multiple vaginal parities have been reported to be an important risk factor for stress urinary incontinence (SUI). Simulated birth trauma with single vaginal distention (VD) has been used to induce the SUI condition in animals; however, the effect of multiple simulated birth traumas on the urethral continence function has not been well characterized. Therefore, we examined the effects of multiple VDs on urethral functions in vivo and the changes in gene expressions of several molecules in the urethra using female SD rats, which were divided into three groups; sham, VD-1 (single VD), and VD-3 groups (3 times of VDs every 2 wk). Two weeks after the final VD, leak point pressure (LPP) and urethral responses during sneezing were evaluated. Also, changes in mRNA levels of urethral molecules were quantified with RT-PCR. The VD-1 group did not show any change in LPP with only a tendency of decrease in amplitudes of the urethral responses during sneezing (A-URS); however, the VD-3 group showed a significant decrease in LPP and urethral responses such as baseline urethral pressure and A-URS accompanied with SUI episodes during sneezing. Nicotinic receptor subtypes and transforming growth factor (TGF)-β1 were significantly increased in both VD-1 and VD-3 groups while TNF receptor (TNFR)-1, IL-6, collagens, and matrix metalloproteinases-9 were significantly increased only in the VD-3 group. These data indicate that rats with multiple simulated birth traumas exhibit profound impairment of the urethral continence function and that these functional changes are associated with those in cytokines, extracellular matrix molecules, and nicotinic receptor subtypes in the urethra.

Balloon pneumo-occluder for vaginal distension at the time of high dose rate (HDR) brachytherapy in patients with cervical cancer

Balloon pneumo-occluder for vaginal distension at the time of high dose rate (HDR) brachytherapy in patients with cervical cancer

PD50-09 A POTENTIAL NEW TARGET FOR STRESS URINARY INCONTINENCE: A μ-OPIOID RECEPTOR IN THE SPINAL CORD BY TRAMADOL, IN RATS

You have accessJournal of UrologyUrodynamics/Lower Urinary Tract Dysfunction/Female Pelvic Medicine: Female Incontinence: Therapy II1 Apr 2017PD50-09 A POTENTIAL NEW TARGET FOR STRESS URINARY INCONTINENCE: A μ-OPIOID RECEPTOR IN THE SPINAL CORD BY TRAMADOL, IN RATS Asuka Ashikari, Minoru Miyazato, Takuma Oshiro, and Seiichi Saito Asuka AshikariAsuka Ashikari More articles by this author , Minoru MiyazatoMinoru Miyazato More articles by this author , Takuma OshiroTakuma Oshiro More articles by this author , and Seiichi SaitoSeiichi Saito More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.2221AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Stress urinary incontinence (SUI) is the most common type of urinary incontinence in women. As the efficacy of pharmacotherapy for SUI is generally unsatisfactory, surgery seems to be the best option for achieving long-term continence. Thus, new effective drugs for SUI should be developed. Tramadol is widely used as an analgesic. It combines the effects of μ-opioid receptors with inhibition of serotonin and noradrenaline reuptake inhibitors. Both effects may be useful for treatment of SUI, but the efficacy of tramadol for SUI has not been examined yet. We previously established a rat model that can be used for examining the active urethral closure mechanism during sneezing. We therefore investigated the effect of tramadol on urethral continence reflex by using this model. METHODS Healthy female rats and rats with SUI induced by vaginal distension (VD) were used. (1) The effects of tramadol on tilt leak point pressure (tilt LPP) were examined before and after intravenous (i.v.) injection of tramadol under urethane anesthesia in both rats. (2) To investigate the effect of tramadol on mid-urethral responses during sneezing, the amplitude of the urethral responses during sneezing (A-URS) and urethral baseline pressure (UBP) were measured by inserting a microtip transducer catheter in the middle urethra in both rats. (3) To confirm the effects of tramadol on μ-opioid receptors in the spinal cord, the effect of tramadol in the presence of intrathecal (i.t.) cyprodime, a selective μ-opioid receptor antagonist, was examined in the healthy rats. RESULTS (1) Tilt LPP was lower in the VD rats than in the healthy rats (43.9 vs 48.6 cmH2O). Tramadol (i.v.) significantly increased the tilt LPP by 25.9% and 21.2% in the healthy and VD rats, respectively. (2) In the healthy rats, tramadol (i.v.) enhanced the A-URS and UBP by 37.0% and 74.9%, respectively. In the VD rats, tramadol administration also enhanced UBP by 11.1% but did not affect A-URS. (3) Administration of cyprodime alone (i.t.) did not affect A-URS and UBP. However, in the presence of cyprodime, tramadol-induced increases in A-URS were suppressed, while UBP was elevated significantly. CONCLUSIONS These results indicate that tramadol, a μ-opioid agonist, is effective for enhancing the active urethral continence reflex during sneezing at the spinal level (a microtip transducer catheter measurement), thereby preventing SUI (a LPP measurement). Therefore, activation of μ-opioid receptors in the spinal cord may be a new candidate treatment for SUI in humans. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e982-e983 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Asuka Ashikari More articles by this author Minoru Miyazato More articles by this author Takuma Oshiro More articles by this author Seiichi Saito More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Therapeutic Effect and Mechanism of Electrical Stimulation in Female Stress Urinary Incontinence

To investigate the therapeutic effect and underlying molecular mechanism of electrical stimulation (ES) in a murine stress urinary incontinence (SUI) model. Sixty female C57BL/6 mice were divided into 4 groups: CON group, no intervention; VD group, vaginal distension (VD) with an 8-mm dilator for 1 hour; VD + ES 20 group, 20 Hz ES for 0.5 hour for 7 days after VD; and VD + ES 50 group, 50 Hz ES for 7 days after VD. For functional studies, assessments of urodynamics and sneezing test were performed; then, anterior vaginal wall specimens were collected. Pathological changes were validated by Masson's trichrome and Van Gieson staining, and the expressions of collagen, transforming growth factor (TGF)-β1-Smad2/3 pathway components, and T-type calcium channels were detected by Western blotting and reverse transcription polymerase chain reaction. ES significantly increased maximum bladder capacity, leak point pressure, and sneezing positive rate in SUI mice. The staining results showed that collagen was disorganized in the VD group but became organized after ES, especially at 50 Hz. The same results were found for collagens I and III. The expression of TGF-β1, p-Smad2 and p-Smad3 significantly decreased in the VD group and significantly increased in the VD + ES groups, especially in the VD + ES 50 group. The expression of 2 T-type calcium channel subtypes (Cav 3.1 and Cav 3.2) decreased in the VD group compared with the CON group, but increased in the VD + ES group compared with the VD group. Dysregulation of collagen metabolism is involved in the pathogenesis of SUI. ES can ameliorate the symptoms of SUI by activating collagen regeneration through the TGFβ1-Smad2/3 pathway. T-type calcium channels might be involved in these processes.

Potential therapeutic role of punicalagin against mechanical-trauma-induced stress urinary incontinence via upregulation of Nrf2 and TGF-β1 signaling

Introduction and hypothesisWe investigated the effect of punicalagin (PUN; 2,3-hexahydroxydiphenoyl-gallagyl-D-glucose), on mechanical-trauma-induced stress urinary incontinence (SUI) in mouse and the mechanisms underlying any effects.MethodsNinety virgin female C57BL/6 mice were randomized into six groups: five groups underwent vaginal distention (VD) for 1 h and leak-point pressure (LPP) was measured on the 1st, 3rd, 7th, 14th, and 28th day following (VD groups 1 d, 3 d, 7 d, 14 d, and 28 d). The sixth group was a noninstrumented control (NC) group. Then, 75 virgin female C57BL/6 mice were randomized into five groups: a VD group (that just underwent VD) and an NC group were orally administered saline every day for 7 days; and three VD + PUN groups that underwent VD and were orally administered PUN respectively at 2.5, 5, and 10 mg/kg every day for 7 days. LPP was tested on the day 7, then all mice were sacrificed and their urethras and anterior vaginal walls harvested for Masson staining, immunohistochemistry study, Western blot analysis, and quantitative polymerase chain reaction (qPCR).ResultsLPPs after VD were significantly lower than the NC group, and the LPPs of mice on days 14 and 28 day after VD were significantly higher than on the days 1, 3, and 7. PUN significantly improved VD-induced drops in LPP and alleviated VD-induced decrease of collagen I, collagen III, α-smooth muscle actin (SMA), transforming growth factor (TGF)-β1, and p-Smad3, nuclear factor-erythroid 2 p45-related factor 2 (Nrf2), and glutathione peroxidase (GPx1) protein levels, and increase of 8-hydroxydeoxyguanosine (OHdG) in urethra and anterior vaginal wall. PUN also up-regulated the expression of manganese superoxide dismutase (MnSOD), whereas protein levels of Smad 2, p-Smad2, and Smad3 were not changed.ConclusionsPUN exerts certain therapeutic effect on mechanical-trauma-induced SUI in mice, which might be through the activation of TGF-β1/Smad3 and Nrf2/antioxidant response element (ARE) signaling activation.

Comparison and analysis of three types of stress urinary incontinence mice models

Objective To compare the 3 kinds of stress urinary incontinence (SUI) mice models. Methods 140 female mice were randomly divided into 4 groups: control group, vaginal distension (VD) group, ovariectomy (OV) group, and VD+ OV group. There were 65 mice in control group, and 25 mice each in the rest three groups. The postoperative infection rate and mortality rate were recorded, and maximum bladder capacity (MBC) and leak point pressures (LPPs) were measured. The contents and activity of matrix metalloproteinase (MMP)-2 and MMP-9 were detected by zymography assay after surgery. The advantages and disadvantages of the three models were comparatively analyzed. Results The MBC/abdominal leak point pressure (ALPP)/bladder leak point pressure (BLPP) in control group, VD group, OV group, and VD+ OV group were respectively (0.101±0.014) ml/(46.420±6.399) cmH2O/(44.180±4.912) cmH2O (1 cmH2O=0.098 kPa); (0.081±0.015) ml/(30.990±5.021) cmH2O /(26.680±6.485) cmH2O; (0.095±0.010) ml/(44.330±6.465) cmH2O /(41.450±6.474) cmH2O and (0.071±0.013) ml/(28.920±5.232) cmH2O /(22.120±7.412) cmH2O, reduced significantly in VD group (P value respectively were 0.000, 0.000 and 0.000) and VD+ OV group (P value respectively were 0.000, 0.000 and 0.000), compared with control group and OV group, but the difference between VD+ OV group and VD group was not notable (P value respectively were 0.051, 0.245 and 0.062), or between OV group and control group (P value respectively were 0.093, 0.238 and 0.064); The expression of MMP-2/MMP-9 in these group were respectively 13.930±0.823/9.610±0.771, 14.990±0.908/11.100±0.735, 13.890±0.707/9.761±0.680, 15.470±1.070/11.240±0.750, increased notably in VD group (P value respectively were 0.003/0.000 and 0.001/0.000) and VD+ OV group (P value respectively were 0.000/0.000 and 0.000/0.000) compared with control grou, but the difference of MMP-2/MMP-9 between the VD+ OV group and VD group was not significant (P value respectively were 0.216/0.604), or between OV group and control group (P value respectively were 0.912/0.573). Conclusion Vaginal distension and vaginal distension with ovariectomy can both establish SUI mice models, but vaginal distension is preferable. Key words: Stress Urinary Incontinence; Vaginal Distension; Ovariectomy; Urine Dynamics Measurement

Mechanism of Mechanical Trauma-Induced Extracellular Matrix Remodeling of Fibroblasts in Association with Nrf2/ARE Signaling Suppression Mediating TGF-β1/Smad3 Signaling Inhibition.

Stress urinary incontinence (SUI) is a common hygienic problem affecting the quality of women's life worldwide. In this research, we revealed the involvement and regulation of extracellular matrix (ECM) remodeling, oxidative damage, and TGF-β1 signaling in the pathological mechanisms of mechanical trauma-induced SUI. We found that excessive mechanical strain significantly increased apoptosis rate, decreased cell viability and ECM production, and broke the balance of MMPs/TIMPs compared with the nonstrain control (NC) group. The expression levels of TGFβ1, p-Smad3, Nrf2, GPx1, and CAT were downregulated, the production of ROS, 8-OHdG, 4-HNE, and MDA was increased, and the nuclear translocation of Smad2/3 was suppressed after 5333 μstrain's treatment. Both mTGF-β1 pretreatment and Nrf2 overexpression could reverse mechanical injury-induced TGFβ1/Smad3 signaling inhibition and ECM remodeling, whereas mTGF-β1 had no effect on Nrf2 expression. Nrf2 overexpression significantly alleviated mechanical injury-induced ROS accumulation and oxidative damage; in contrast, Nrf2 silencing exhibited opposite effects. Besides, vaginal distention- (VD-) induced in vivo SUI model was used to confirm the in vitro results; Nrf2 knockout aggravates mechanical trauma-induced LPP reduction, ECM remodeling, oxidative damage, and TGF-β1/Smad3 suppression in mice. Therefore, we deduce that mechanical injury-induced ECM remodeling might be associated with Nrf2/ARE signaling suppression mediating TGF-β1/Smad3 signaling inhibition. This might reflect a new molecular target for SUI researches.

AB303. SPR-30 Neurotrophin dysregulation after dual injury childbirth model is corrected via electrical stimulation of the pudendal nerve in a rat model

ObjectiveStress urinary incontinence (SUI), the leakage of urine due to an increase in abdominal pressure, affects 30% of women over the age of 40. Childbirth is the primary risk factor for SUI since the child’s head passing through the birth canal can injure the EUS and the nerve that innervates it, the pudendal nerve (PN). Animal models with combined EUS [vaginal distension (VD)] and PN crush (PNC) injuries have shown that the nerve does not recover as well as with PNC alone and the animals have delayed return of function. Typically after a nerve injury, the neuron upregulates neurotrophin factors [i.e., brain derived neurotrophin factor (BDNF)] and their receptors [i.e., tyrosine kinase B (Trk B) and p75], and Schwann cells also upregulate neurotrophin factors [i.e., glial derived neurotrophin factor (GDNF) and NT4] as part of the regeneration process. The impaired nerve regeneration in the dual injury model was shown to be due to a dysregulation of BDNF in the PN motoneuron, so any potential treatment would need to upregulate BDNF. Electrical stimulation (ES) has been shown to upregulate BDNF and Trk B in neurons and GDNF in Schwann cells. Our previous work has shown daily and 4 times per week stimulation for 2 weeks accelerates functional recovery after PNC + VD. The aim of the study was to determine if daily stimulation causes an upregulation of neurotrophin factor and their receptors in motoneurons of the PN after injury. We hypothesize that the accelerated recovery from ES is mediated by an upregulation of BDNF and NT4 and their receptor Trk B, as well as GFRα1, the receptor of GDNF.MethodsAnimals were divided into five groups: Sham PNC, PNC, PNC + VD, PNC + VD with sham stimulation (SS) or PNC + VD + ES. ES (20 Hz, 0.3 mA, 0.1 ms) was conducted daily for 1 or 2 weeks after injury, when the spinal cord was harvested from L4–S3. The spinal cord was sectioned and Onuf’s nucleus, the motoneuron nucleus for the PN, was harvested via laser capture microscopy. RNA was then isolated and preamplified before running PCR and compared to GAPDH expression. A one way Anova was used to determine statistically significant differences between groups.ResultsOne week after PNC + VD, BDNF was not upregulated as previously shown. Neither were p75 or GFRα1, as expected. NT4 was not downregulated after PNC + VD, as expected. P75 was significantly increased in the PNC + VD + ES group compared to the PNC + VD group 2 weeks after injury. BDNF was also upregulated 2 weeks after PNC + VD + ES compared to PNC + VD.ConclusionsP75, NT4 and GFRα1 expression data suggest that PNC + VD causes dysregulation of other regenerative proteins other than BDNF. BDNF was upregulated 2 weeks after PNC + VD with ES. Future research will be designed to investigate the mechanism of this effect.Funding Source(s)VA Merit A1262-R and the RR&D Service of the VA

Clinical Implications of Obstructed Hemivagina and Ipsilateral Renal Anomaly (OHVIRA) Syndrome in the Prepubertal Age Group.

PurposeObstructed hemivagina and ipsilateral renal anomaly (OHVIRA) syndrome is a rare syndrome characterized by Müllerian duct and renal anomalies. It is usually regarded as a disease of adolescence; however, due to a number of possible problems, the management of patients before puberty should not be overlooked. We assessed the clinical course of prepubertal patients to propose appropriate management.Materials and MethodsWe retrospectively assessed 43 prepubertal OHVIRA syndrome patients who were diagnosed and followed up at our institution from July 2004 to June 2015. We reviewed medical records, focusing on presentation, radiologic findings, surgical management, and the overall clinical course.ResultsMedian age at diagnosis was 1.3 months and median follow-up period was 25.5 months. The most common accompanying ipsilateral urologic anomalies were ectopic ureter and ureterocele, while the most common contralateral anomaly was vesicoureteral reflux. During the follow-up period, six patients (14.0%) required surgery at a median age of 31.2 months due to recurrent urinary tract infection, uncontrolled vaginal distention compressing adjacent organs, urinary incontinence, or intractable abdominal pain.ConclusionsWhile OHVIRA syndrome is known as a postpubertal disease, about 13% of prepubertal patients in our study required surgery. When ectopic ureter insertion into the vagina is present, further treatment may be needed to address the complications caused by continuous urine production. Patients should be monitored for complications arising from either obstructed hemivagina or renal anomalies with regular follow-up, especially before the age of five years.

Exploring the dynamics of progenitor cells in the urethra after simulated birth trauma in mice

ObjectiveTo examine the alteration in the cellular dynamics of the urethral tissue after a simulated birth trauma in a mouse model. Materials and MethodsA total of 36 B6 mice received vaginal distention treatment, and four untreated mice were used as controls. Specimens were collected every 24 hours after the injury for 9 consecutive days and examined using immunofluorescent staining for cell markers including c-kit, smooth muscle actin (SMA), and vimentin. Confocal microscopy was used to localize the stained cells and determine the cell number. ResultsThe number of c-kit positive cells increased after the 1st day and peaked on the 3rd day. The amount of SMA positive cells rapidly reduced to its lowest count on the 1st day and maintained a statistically significant low cell number than that at the basal level for 4 days after vaginal distension. The cell number finally returned to basal level on the 9th day. The amount of vimentin positive cells increased dramatically after the 1st day and plateaued from the 3rd day to the 9th day. The number of vimentin positive cells in the plateau phase was significantly higher than that of the control group. ConclusionOur study confirmed that the dynamic change in different cell types after the urethral injury was dependent on the nature and physiology of the wound repairing cells during the tissue healing process. It might be a simple animal model to study birth trauma repair; however, the varied progenitor cell activity in different species should also be considered.

Local release from affinity-based polymers increases urethral concentration of the stem cell chemokine CCL7 in rats

The protein chemokine (C-C motif) ligand 7 (CCL7) is significantly over-expressed in urethral and vaginal tissues immediately following vaginal distention in a rat model of stress urinary incontinence. Further evidence, in this scenario and other clinical scenarios, indicates CCL7 stimulates stem cell homing for regenerative repair. This CCL7 gradient is likely absent or compromised in the natural repair process of women who continue to suffer from SUI into advanced age. We evaluated the feasibility of locally providing this missing CCL7 gradient by means of an affinity-based implantable polymer. To engineer these polymers we screened the affinity of different proteoglycans, to use them as CCL7-binding hosts. We found heparin to be the strongest binding host for CCL7 with a 0.323 nM dissociation constant. Our experimental approach indicates conjugation of heparin to a polymer backbone (using either bovine serum albumin or poly (ethylene glycol) as the base polymer) can be used as a delivery system capable of providing sustained concentrations of CCL7 in a therapeutically useful range up to a month in vitro. With this approach we are able to detect, after polymer implantation, significant increase in CCL7 in the urethral tissue directly surrounding the polymer implants with only trace amounts of human CCL7 present in the blood of the animals. Whole animal serial sectioning shows evidence of retention of locally injected human mesenchymal stem cells (hMSCs) only in animals with sustained CCL7 delivery, 2 weeks after affinity-polymers were implanted.

PD35-05 HUMAN URINE-DERIVED STEM CELLS OR THEIR SECRETOME ALONE FACILITATE FUNCTIONAL RECOVERY IN A RAT MODEL OF STRESS URINARY INCONTINENCE

You have accessJournal of UrologyStem Cell Research1 Apr 2016PD35-05 HUMAN URINE-DERIVED STEM CELLS OR THEIR SECRETOME ALONE FACILITATE FUNCTIONAL RECOVERY IN A RAT MODEL OF STRESS URINARY INCONTINENCE Christine Tran, Abhi Tangada, Hualin Yi, Brian Balog, Yuanyuan Zhang, and Margot Damaser Christine TranChristine Tran More articles by this author , Abhi TangadaAbhi Tangada More articles by this author , Hualin YiHualin Yi More articles by this author , Brian BalogBrian Balog More articles by this author , Yuanyuan ZhangYuanyuan Zhang More articles by this author , and Margot DamaserMargot Damaser More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.1077AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Stem cell based therapy has emerged as a promising alternative to current female stress urinary incontinence (SUI) treatments. Human urine-derived stem cells (USCs) have no risk from biopsy but have not previously been investigated for SUI. One therapeutic mechanism of action of stem cells is secretion of bioactive growth factors or ″secretome″ that drives the regulation of many physiological processes via paracrine actions. We hypothesized that USCs or their secretome alone would promote functional recovery in a rodent model of female SUI. METHODS Thirty two-female Sprague-Dawley rats were randomized into 3 groups and underwent vaginal distension (VD) followed by intraperitoneal delivery of USCs (VD + USC), VD and saline, or sham VD and saline 1 hour after injury. To investigate if factors secreted by USCs facilitate functional recovery after VD, concentrated conditioned media (CCM) was generated by incubating confluent USCs in serum-free media for 24 hours. Cultured supernatant was then extracted, washed, and concentrated to form CCM. Concentrated media (CM) was created by concentrating serum-free media without the influence of cells. Thirty rats randomized into 3 groups underwent VD and intraperitoneal delivery of CCM (VD + CCM), VD and CM, or sham VD and CM 1 hour after injury. One week after injury, treatment efficacy was assessed by measurement of leak point pressure (LPP), and histological and immunocytochemical analysis of the urethra. RESULTS After 1 week, LPP significantly increased after VD in rats treated with USCs or CCM, compared to animals that received saline, and was similar to sham VD animals. On semiquantitative histological analysis, there was no significant difference between both the striated and smooth muscle components of the urethral sphincter of VD + USC and VD + CCM animals compared to sham VD animals; VD + saline and VD + CM animals, however, demonstrated significant muscle fiber attenuation and disorganization. Additionally, elastin fibers in VD + USC and VD + CCM animals were long, thickened, and mostly oriented compared to the short, thin, and disoriented fibers in VD + saline and VD + CM animals. No implanted USCs were found in the region of urethra. CONCLUSIONS Intraperitoneal injection of USCs and their secretions facilitate recovery from SUI in a rat model, likely via paracrine effects. Furthermore, elastogenesis may play a role in recovery of urethral function. USCs represent an attractive, alternate stem cell source with no biopsy risk to target the underlying pathophysiology in female SUI. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e845 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Christine Tran More articles by this author Abhi Tangada More articles by this author Hualin Yi More articles by this author Brian Balog More articles by this author Yuanyuan Zhang More articles by this author Margot Damaser More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

MP65-01 STROMAL CELL DERIVED FACTOR-1 ACCELERATES RECOVERY OF CONTINENCE IN RAT MODEL OF VAGINAL DISTENSION INJURY

You have accessJournal of UrologyUrodynamics/Lower Urinary Tract Dysfunction/Female Pelvic Medicine: Female Incontinence: Therapy II1 Apr 2016MP65-01 STROMAL CELL DERIVED FACTOR-1 ACCELERATES RECOVERY OF CONTINENCE IN RAT MODEL OF VAGINAL DISTENSION INJURY Ahmad Khalifa, Amr Mahran, Michael Kavran, Juliana Woda, Marc Penn, and Adonis Hijaz Ahmad KhalifaAhmad Khalifa More articles by this author , Amr MahranAmr Mahran More articles by this author , Michael KavranMichael Kavran More articles by this author , Juliana WodaJuliana Woda More articles by this author , Marc PennMarc Penn More articles by this author , and Adonis HijazAdonis Hijaz More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.1212AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Vaginal distension (VD) in rats results in a reproducible injury to urethral continence mechanism that is recoverable in 2 weeks. VD injury has been shown to cause ischemia. Stromal cell derived factor-1 (SDF-1), is a known chemokine involved in angiogenesis and chemo attraction of stem cells into injury area. Previously in animal and human studies, delivery of non-viral human SDF-1 plasmid (pSDF-1), which expresses SDF-1 for 2-3 weeks, has shown safety, increased vessel density and improved patient status in ischemic heart and vascular diseases. In this experiment we examined the efficacy of pSDF-1 injection on continence recovery after VD & correlated the changes with histological changes. METHODS A total of 72 Sprague Dawley rats underwent VD and were randomly divided into 2 groups. Group 1 (42 rats) underwent SDF-1 injection followed by Leak point pressure (LPP) measurement at 4, 7, and 14 days (14 rats per time point). Group 2 (30 rats) were sham injected (5% dextrose) after VD and LPP was measured at same time points (10 rats per time point). VD was done as described with inflation of a 10 French modified Foley catheter intravaginal for 4 hours to create injury. SDF-1 and dextrose were injected peri-uretherally immediately after VD. Following LPP measurement, urogenital tissues were harvested for histological examination. Slides were stained with H&E and Masson’s trichrome. Vascularity and collagen content was determined using Adobe Photoshop and Image J software. Data analysis was done by IBM SPSS v.22. RESULTS As expected, the LPP decreased at 4 days in both groups. SDF-1 group exhibited a significant increase in LPP at 7 days compared to the sham (22.3 cmH2O vs 13.62 3 cmH2O, P value <0.001 respectively). The improvement in LPP continued in both groups at day 14, with the statistical difference approaching significance (36.38 cmH2O vs 25.82 cmH2O respectively, p=0.067). Histological evaluation and image analysis revealed no gross differences in the smooth and skeletal muscle thickness of the mid-urethra with no statistically significant difference in the collagen content and vascular density in the urethra at all-time points. CONCLUSIONS pSDF-1 plasmid injection at the time of VD accelerated the recovery of continence in the rat model of simulated birth trauma. The mechanism of improved recovery is not evident from the gross histological evaluation. Further research is needed to explain the mechanism of early improvement, possible hypothesizes are stem cell attraction and decrease in cellular injury or death. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e864 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Ahmad Khalifa More articles by this author Amr Mahran More articles by this author Michael Kavran More articles by this author Juliana Woda More articles by this author Marc Penn More articles by this author Adonis Hijaz More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

MP68-03 EVALUATION OF CANDIDATE URINARY BIOMARKERS FOR UROLOGIC CHRONIC PELVIC PAIN SYNDROME (UCPPS)

INTRODUCTION AND OBJECTIVES: Stress urinary incontinence, a major type of urinary incontinence, increases with age and is often developed after partum injury. Low intensity pulsed ultrasound (LIPUS) has been investigated in the treatment of many diseases showing its ability of restoring soft tissue injury. We investigated the therapeutic effect of low intensity pulsed ultrasound in stress urinary incontinence. METHODS: Thirty-two Sprague Dawley rats in SUI group underwent vaginal distension (VD) and bilateral ovariectomy mimicking partum injury. Eight rats served as mock operation control. Eight rats each in SUI group was treated with low-dosage LESW (0.03 mJ/mm2), medium-dosage LESW (0.06 mJ/mm2), or high-dosage LESW (0.09 mJ/mm2). The rest eight rats served as none-treatment group. For functional study, leak point pressure test (LPP) was performed 2 weeks after the last LESW. Masson trichrome staining was performed to validate the pathological changes. RESULTS: The LPP was restored in medium-dosage LESW and high-dosage LESW groups, but not in low-dosage LESW group. More robust striated muscle regeneration was found in these two group comparing with the none-treatment group. CONCLUSIONS: LIPUS ameliorate the symptom of SUI via activating striated muscle regeneration.

MP17-19 CELL THERAPY FOR NEUROGENIC DETRUSOR OVERATIVITY RESULTING FROM CHRONIC SPINAL CORD INJURY (SCI) IN RATS

INTRODUCTION AND OBJECTIVES: Stress urinary incontinence, a major type of urinary incontinence, increases with age and is often developed after partum injury. Low intensity pulsed ultrasound (LIPUS) has been investigated in the treatment of many diseases showing its ability of restoring soft tissue injury. We investigated the therapeutic effect of low intensity pulsed ultrasound in stress urinary incontinence. METHODS: Thirty-two Sprague Dawley rats in SUI group underwent vaginal distension (VD) and bilateral ovariectomy mimicking partum injury. Eight rats served as mock operation control. Eight rats each in SUI group was treated with low-dosage LESW (0.03 mJ/mm2), medium-dosage LESW (0.06 mJ/mm2), or high-dosage LESW (0.09 mJ/mm2). The rest eight rats served as none-treatment group. For functional study, leak point pressure test (LPP) was performed 2 weeks after the last LESW. Masson trichrome staining was performed to validate the pathological changes. RESULTS: The LPP was restored in medium-dosage LESW and high-dosage LESW groups, but not in low-dosage LESW group. More robust striated muscle regeneration was found in these two group comparing with the none-treatment group. CONCLUSIONS: LIPUS ameliorate the symptom of SUI via activating striated muscle regeneration.

MP30-01 IDENTIFYING A SENSITIVE AND SPECIFIC PANEL OF DNA METHYLATION MARKERS IN MODELS OF CYSTITIS

stimulation of the pudendal nerve upregulates neurotrophin expression and improves recovery in a rat model of SUI. METHODS: Rats received 1 hour of electrical stimulation (20 Hz, 0.3 mA, 0.1 ms pulse) or sham stimulation of the pudendal nerve immediately after simulated birth injury, consisting of vaginal distension and pudendal nerve crush. A separate cohort of rats received sham injury and no treatment. All rats underwent stimulation or sham stimulation for 1 hour twice per week for 2 weeks. Brain derived neurotrophic factor (BDNF) and bII-tubulin (neuronal cytoskeletal protein and marker of neuroregenerative response) expression in Onuf’s nucleus was measured with RT-PCR. Leak point pressure (LPP) and external urethral sphincter electromyography (EUS EMG) was performed. Data was analyzed with a two-way ANOVA and Student-Newman-Keuls posthoc pairwise comparisons with p<0.05 statistically significant. RESULTS: Compared to sham stimulation, electrical stimulation significantly increased both BDNF and bII-tubulin expression in Onuf’s nucleus, demonstrating an increased neuroregenerative response. Both LPP and baseline EUS EMG amplitude decreased significantly after injury compared to sham injury and increased significantly with stimulation compared to sham stimulation. During voiding, EUS EMG burst frequency, burst duration, and interburst interval were significantly decreased by injury compared to sham and after injury were significantly increased with electrical stimulation compared to sham. CONCLUSIONS: Pudendal nerve electrical stimulation appears to promote SUI recovery via strengthening the neuroregenerative response by upregulating BDNF expression in pudendal nerve cell bodies. Electrical stimulation may shorten postpartum SUI duration and could potentially serve as a preventative treatment for later SUI development.