Myostatin Overexpression and Smad Pathway in Detrusor Derived from Pediatric Patients with End-Stage Lower Urinary Tract Dysfunction.

Cell therapies and tissue engineering approaches using smooth muscle cells (SMCs) may provide treatment alternatives for end-stage lower urinary tract dysfunction (ESLUTD). Myostatin, a negative regulator of muscle mass, is a promising target to improve muscle function through tissue engineering. The ultimate goal of our project was to investigate the expression of myostatin and its potential impact in SMCs derived from healthy pediatric bladders and pediatric ESLUTD patients. Human bladder tissue samples were evaluated histologically, and SMCs were isolated and characterized. The proliferation of SMCs was assessed by WST-1 assay. The expression pattern of myostatin, its pathway and the contractile phenotype of the cells were investigated at gene and protein levels by real-time PCR, flow cytometry, immunofluorescence, WES and gel contraction assay. Our results show that myostatin is expressed in human bladder smooth muscle tissue and in isolated SMCs at gene and protein levels. A higher expression of myostatin was detected in ESLUTD-derived compared to control SMCs. Histological assessment of bladder tissue confirmed structural changes and decreased muscle-to-collagen ratios in ESLUTD bladders. A decrease in cell proliferation and in the expression of key contractile genes and proteins, α-SMA, calponin, smoothelin and MyH11, as well as a lower degree of in vitro contractility was observed in ESLUTD-derived compared to control SMCs. A reduction in the myostatin-related proteins Smad 2 and follistatin, and an upregulation in the proteins p-Smad 2 and Smad 7 were observed in ESLUTD SMC samples. This is the first demonstration of myostatin expression in bladder tissue and cells. The increased expression of myostatin and the changes in the Smad pathways were observed in ESLUTD patients. Therefore, myostatin inhibitors could be considered for the enhancement of SMCs for tissue engineering applications and as a therapeutic option for patients with ESLUTD and other smooth muscle disorders.

Improved contractile potential in detrusor microtissues from pediatric patients with end stage lower urinary tract dysfunction.

Autologous cell-based tissue engineering has been proposed as a treatment option for end stage lower urinary tract dysfunction (ESLUTD). However, it is generally accepted that cells isolated from patient bladders retain the pathological properties of their tissue of origin and therefore need to be improved before they can serve as a cell source for tissue engineering applications. We hypothesize that human three-dimensional (3D) microtissues of detrusor smooth muscle cells (SMCs) are valuable ex vivo disease models and potent building blocks for bladder tissue engineering. Detrusor SMCs isolated from bladder wall biopsies of pediatric ESLUTD patients and healthy controls were expanded and cultured into 3D microtissues. Gene and protein analyses were performed to explore the effect of microtissue formation on SMC viability, contractile potential, bladder wall specific extracellular matrix (ECM) composition and mediators of ECM remodeling. Through microtissue formation, remodeling and intensified cell-cell interactions, the ESLUTD SMCs lost their characteristic disease phenotype. These microtissues exhibited similar patterns of smooth muscle related contractile proteins and essential bladder wall-specific ECM components as microtissues from healthy control subjects. Thus, the presented data suggest improved contractile potential and ECM composition in detrusor SMC microtissues from pediatric ESLUTD patients. These findings are of great relevance, as 3D detrusor SMC microtissues might be an appropriate cell source for autologous cell-based bladder tissue engineering.

Seroprevalence of SARS-CoV-2 antibodies, associated factors, experiences and attitudes of nursing home and home healthcare employees in Switzerland.

BACKGROUND: Many studies in hospital settings exist and have shown healthcare employees to be particularly exposed to SARS-CoV-2. While research focused on hospital staff, little evidence exists for employees in nursing homes and home care. The aims of this study were to assess the seroprevalence in nursing homes and home care employees in the Canton of Zurich, compare it to the general population, assess factors associated with seropositivity and explore the perspective of the employees regarding how the pandemic changed their daily work. METHODS: This cross-sectional study is part of the national Corona Immunitas research program of coordinated, seroprevalence studies in Switzerland. Six nursing homes and six home healthcare organizations providing at home care services in Zurich were selected and 296 and 131 employees were recruited, respectively. Assessments included standardized questionnaires, blood sampling for antibodies, and additional work-specific questions. All participants were recruited between 21st September and 23rd October 2020, before the second wave of the pandemic hit Switzerland, and were possibly exposed to SARS-CoV-2 at their work during the first wave in spring 2020. RESULTS: Seroprevalence of SARS-CoV-2 was 14.9% (95% CI 11.1%-19.6%; range 3.8% to 24.4%) for nursing home employees and 3.8% (95% CI 1.4-9.1%; range 0% to 10%) for home healthcare employees, compared to the general population of Zurich at 3.5% in September 2020 for those aged 20-64. Nurses were 2.6 times more likely to have SARS-CoV-2 antibodies than those employees who were not nurses (95% CI 1.1-6.2). The employees (nursing homes vs. home healthcare) perceived the implementation of general safety measures (44.9% vs. 57.3%) and wearing masks during work (36.8% vs. 43.5%), especially due to the limited communication with residents/clients, as the most crucial changes. CONCLUSIONS: Nursing home employees who worked through SARS-CoV-2 outbreaks at their work were substantially more affected by SARS-CoV-2 infection compared to the general population and to home healthcare employees who similarly worked through outbreaks in their communities. Employees reported that important resources to cope with the burdensome changes they perceived in their daily work were personal resources and team support. TRIAL REGISTRATION: Current Controlled Trials ISRCTN18181860 dated 09/07/2020. Retrospectively registered.