Secretomes of human pluripotent stem cell-derived smooth muscle cell progenitors upregulate extracellular matrix metabolism in the lower urinary tract and vagina

Background Adult mesenchymal stem cells (MSCs) have been studied extensively for regenerative medicine; however, they have limited proliferation in vitro, and the long culture time induces cell senescence. MSCs also contribute to tissue repair through their paracrine function. In this study, we sought to examine the paracrine effects of human smooth muscle cell progenitors (pSMC) on the urethra and adjacent vagina of stress urinary incontinence rodents. We use human pluripotent stem cell (PSC) lines to derive pSMCs to overcome the issue of decreased proliferation in tissue culture and to obtain a homogenous cell population. Method Three human PSC lines were differentiated into pSMCs. The conditioned medium (CM) from pSMC culture, which contain pSMC secretomes, was harvested. To examine the effect of the CM on the extracellular matrix of the lower urinary tract, human bladder smooth muscle cells (bSMCs) and vaginal fibroblasts were treated with pSMC-CM in vitro. Stress urinary incontinence (SUI) was induced in rats by surgical injury of the urethra and adjacent vagina. SUI rats were treated with pSMC-CM and monitored for 5 weeks. Urethral pressure testing was performed prior to euthanasia, and tissues were harvested for PCR, Western blot, and histological staining. Kruskal-Wallis one-way ANOVA test and Student t test were used for statistical comparisons. Results pSMC-CM upregulated MMP-2, TIMP-2, collagen, and elastin gene expression, and MMP-9 activity in the human bladder and vaginal cells consistent with elastin metabolism modulation. pSMC-CM treatment in the SUI rat improved urethral pressure (increase in leak point pressure compared to intact controls, p < 0.05) and increased collagen and elastin expression in the urethra and the adjacent vagina. Conclusion Conditioned media from smooth muscle cell progenitors derived from human pluripotent stem cells improved urethral leak point pressure and collagen and elastin content in the SUI rat. These findings suggest a novel therapeutic potential for PSC-based treatments for SUI and pelvic floor disorders where tissues are affected by collagen, elastin, and smooth muscle loss.

Performance of Urodynamics During the COVID-19 Pandemic: A Questionnaire Survey

AimsThe principal aim of this multicentre, international cohort study is to explore the safety concerns of clinicians when performing urodynamic studies (UDS) during the COVID-19 pandemic. This study will also assess provider preference on personal protective equipment (PPE) during UDS and awareness of relevant international guidelines. The outcome is to offer a practical means of reducing the risk of aerosol transmis-sion during UDS, to include a protocol for screening patients, and to consider safer methods of inducing urethral leak-point pressure. MethodsA 14-point, multiple-response, online questionnaire was designed through Survey Monkey. This was distributed to healthcare workers involved in UDS via email, WhatsApp, and Twitter. This included, of varying experience; urologists, urogynaecologists, clinical nurse specialists, and clinical scientists across the United Kingdom (UK), United States of America (USA), Australia, Europe, and Middle-East.ResultsThe study was conducted over two weeks and there were 104 responses to the survey. The majority of respondents were consultants (60%) and 73% of respondents performed UDS regularly. There were 56.7% of respondents who felt that UDS was safe to be re-instated and there were 52.8% who did not feel coughing was safe during the procedure. At the time of the survey, 83.5% were unaware of any national or international urological guidelines on the best practice for urodynamics during this era and 52.8% of clinicians stated they would wear PPE during UDS if it were reinstated. ConclusionThe majority of respondents felt safe to reinstate UDS; however, they felt more official guidance and recommendation is needed. Our proposed protocol can provide a safe and practical approach that keeps healthcare workers and patients safe.

Verification and Defined Dosage of Sodium Pentobarbital for a Urodynamic Study in the Possibility of Survival Experiments in Female Rat

Objectives. To evaluate the effects of pentobarbital dosages on lower urinary tract function and to define an appropriate dosage of sodium pentobarbital that would be suitable for urodynamic studies in which recovery from anesthesia and long term survive were needed for subsequent experiment. Methods. Twenty-four 8-week-old, female, virgin, Sprague-Dawley rats (200-250 g) were used in this study. Rats in study groups received gradient doses of pentobarbital intraperitoneally, and those in the control group received urethane intraperitoneally. External urethral sphincter electromyography (EUS-EMG) was recorded simultaneously during cystometry and leak point pressure tests. The toe-pinch reflex was used to determine the level of anesthesia. Results. Micturition was normally induced in both the urethane group and 32 mg/kg pentobarbital group. However, in groups of 40 mg/kg or 36 mg/kg pentobarbital, micturition failed to be induced; instead, nonvoiding contractions accompanied by EUS-EMG tonic activity were observed. There were no significant differences in leak point pressure or EUS-EMG amplitude or frequency between the urethane and 32 mg/kg pentobarbital groups. Conclusions. This study confirmed significant dose-dependent effects of pentobarbital on lower urinary tract function and 32 mg/kg pentobarbital as an appropriate dosage for recovery urodynamic testing, which enable the achievement of expected essential micturition under satisfactory anesthesia in female rats.

Comparison of urodynamic parameters with respect to neurological levels in post-traumatic spinal cord injury patients

Background: Urodynamic evaluation is mandatory in order to correctly assess and classify bladder dysfunction in spinal cord injury (SCI) patients. Study investigated patterns of neurogenic bladder dysfunction in patients with post traumatic spinal cord injury and assessed the relationship of detrusor leak point pressure with compliance, post void residual urine volume and maximum cystometric capacity.Methods: Eighty six patients with neurogenic bladder secondary to traumatic spinal cord injury (SCI) underwent cystometry with electromyography (EMG). T-test was used to compare detrusor leak point pressure (LPP) between complete and incomplete injury groups. Pearson correlation test was used to seek correlation between detrusor LPP and compliance, post void residual volume (PRV) and maximum cystometric capacity (MCC).Results: Mean detrusor LPP in suprasacral complete injury group, suprasacral incomplete injury group and sacral complete injury was 52±21 cm of H2O, 53±18 cm of H2O and 16±9 cm of H2O respectively. No significant difference in detrusor LPP was found between suprasacral complete and incomplete group on t-Test (p= 0.571068). Significant difference in detrusor LPP was found between suprasacral and sacral group (p= 5.71891E-12). Mean compliance in sacral injury group was 24±16 and in suprasacral complete injury group was 5±6. Mean compliance in suprasacral incomplete injury group was 4±2. Pearson correlation showed negative correlation (r = -0.6918934) between detrusor leak point pressure and compliance (p= 1.2744E-13). Negative correlation (r = -0.311409922) was observed between detrusor leak point pressure and post leak/ void residual urine volume (p= 0.003335033) and between detrusor LPP and maximum cystometric capacity (r = -0.31354), (p= 0.003115).Conclusions: Significant difference in urodynamic parameters exists between sacral and suprasacral injury patients. However there is no significant difference in urodynamic parameters between complete and incomplete injury at suprasacral level.

Mid-Urethral Sling for Stress Incontinence: Does Urodynamic Pressure Reading Affect Post-Operative Outcome?

The aim of our study was to determine the effect of preoperative urodynamic reading of valsalva leak point pressure on the result of mid-urethral sling surgery. From January 2010 to December 2014, a total of 207 patients underwent mid-urethral sling surgery at the Toronto Western Hospital. An incontinence questionnaire was sent to 94 patients who accepted to be involved in the study to examine satisfaction post-surgery. Forty-five patients replied and were included in the analysis. The patients were divided into three groups according to valsalva leak point pressure (< 60, 60-80 and > 80 cm H2O) determined on urodynamic testing. Of the forty-five patients who responded to the questionnaire, seven were excluded for only having had stress testing done and two were excluded due to intermittent catheterization. Thirteen patients had evidence of detrusor overactivity on urodynamic testing. Simple linear regression analysis was done for the three groups of the valsalva leak point pressure values and correlation to satisfaction was found -0.263, -0.236, and -0.148, respectively. In this current study, we could not find a relation between valsalva leak point pressure values and the self-reported satisfaction post-surgical correction.

Role of the serotonergic system in urethral continence reflexes during sneezing in rats

To clarify the role of serotonin (5-HT) in the prevention of stress urinary incontinence (SUI) during sneezing, we investigated the effect of intraperitoneal application of p-chlorophenylalanine (PCPA; a serotonin synthesis inhibitor) and intravenous application of CP-809101 (a 5-HT2C agonist) or LP44 (a 5-HT7 agonist) using female rats, in which the neurally evoked continence reflex during sneezing was examined. Amplitudes of urethral pressure response during sneezing (A-URS), urethral baseline pressure (UBP) at the middle urethra, and sneeze-induced leak point pressure (S-LPP) were measured in normal female adult rats with or without drug administration. PCPA decreased A-URS by 35.1 cmH2O and UBP by 13.3 cmH2O compared with normal rats. In PCPA-administrated rats, CP-809101 increased A-URS by 24.1 cmH2O and UBP by 15.1 cmH2O, and LP44 also increased A-URS by 20.6 cmH2O and UBP by 11.4 cmH2O compared with rats treated with PCPA alone. SUI was observed with S-LPP of 40.1 cmH2O in PCPA-administrated rats, in which CP-809101 and LP44 increased S-LPP by 28.0 and 15.2 cmH2O, respectively, compared with rats treated with PCPA alone. The effects of CP-809101 and LP44 were antagonized by SB-242084 (a selective 5-HT2C antagonist) and SB-269970 (a selective 5-HT7 antagonist), respectively. These results indicate that activation of 5-HT receptors enhances the active urethral closure reflex during sneezing, at least in part via 5-HT2C and 5-HT7 receptors.