Development of a muscle progenitor cell-based therapeutic approach for the treatment of stress urinary incontinence

The urethra serves a dual function by maintaining continence during bladder filling and aiding the release of urine during micturition. Within the urethra, a sphincter region containing both smooth and striated muscle layers normally prevents involuntary leakage of urine. However, patients with stress urinary incontinence lose this ability upon sudden increases in intravesical pressure (i.e. from coughing, straining, etc.). This condition has been associated with a decline in striated muscle, which may be susceptible to direct muscle or associated nerve damage. Cellular uromyoplasty proposes to augment this muscle layer through the transplantation of myogenic progenitors. The goal of this work was to address current deficiencies regarding the isolation and identification of efficient progenitors, and the urethral biomechanical consequences of striated muscle restoration. Both issues are essential for effective clinical implementation of this therapeutic approach.The ability of various progenitor populations to regenerate skeletal (striated) muscle was assessed in a dystrophic mouse model. Both cell surface protein expression and behavioral characteristics were investigated for their potential use as indicators of regenerative efficiency. The results demonstrate the limited utility of surface proteins due to fluctuations in expression and lack of regenerative consistency between directly-isolated and cultured cell populations. Behavioral characteristics related to the ability of cells to maintain a proliferative phenotype under differentiation-inducing conditions appears more promising in this regard, and indicates that in vivo expansion of transplanted cells may be a critical variable in the regeneration process. A new ex vivo method to assess the regional biomechanical function of the intact urethra, under physiologic loading conditions, was introduced and validated. Quantitative characterization and comparison of tissue responses to applied intralumenal pressures was performed in the presence or absence of selected muscle activity. The dominant smooth muscle influence observed suggests that a large degree of striated regeneration may be necessary to impart functional changes in urethra mechanics. Importantly, these results also indicate that muscle fiber orientation may significantly impact urethra closure function. Together, this information will be useful in progressing uromyoplasty therapy toward clinical utility, and aid the broader scientific community investigating myogenic cell transplantation and lower urinary tract function

erectile function after radical prostatectomy

In this study our aim is to increase the understanding of the prostate and related organs anatomy for better continence and erectile function results after urological surgery. Prostate and related organs were dissected from seven cadavers. After dissection, 165 serial sections with 300 mu m thickness were derived at a 100 mu m interval. The histological images were examined and imported to the computer. Three-dimensional (3D) remodeling had been performed. The findings were evaluated into three categories: macroscopic, microscopic and 3D reconstruction. Striated muscle fibers had been detected at the anterior fibromuscular stroma in histological sections. In 3D remodeling, urethra seemed to be a complete functional unit, beginning from the trigone up to the membranous urethra. The neurovascular bundles run under the pelvic fascia on both sides and go through to the bladder neck at 5 and 7 o'clock. Computer remodeling demonstrated that neurovascular structures had a close association with the bladder neck and the seminal vesicle. Computer program made it possible to rotate all 3D-reconstructed figures by 3601 and examine them from all possible angles. All reconstructed structures can be examined together at the same time or one by one. Surgeons must pay special attention to the continence area described as a single unit, beginning from trigone to the membranous urethra, during the surgery. Meticulous dissection of the neurovascular bundles, especially close to the seminal vesicles and bladder neck, during the radical prostatectomy is necessary. These reconstructions can be used for the educational purpose of medical students as well as the urology surgeons

Attenuation of leukocyte sequestration by selective blockade of PECAM-1 or VCAM-1 in murine endotoxemia

Background: Molecular mechanisms regulating leukocyte sequestration into the tissue during endotoxemia and/or sepsis are still poorly understood. This in vivo study investigates the biological role of murine PECAM-1 and VCAM-1 for leukocyte sequestration into the lung, liver and striated skin muscle. Methods: Male BALB/c mice were injected intravenously with murine PECAM-1 IgG chimera or monoclonal antibody (mAb) to VCAM-1 ( 3 mg/kg body weight); controls received equivalent doses of IgG2a ( n = 6 per group). Fifteen minutes thereafter, 2 mg/kg body weight of Salmonella abortus equi endotoxin was injected intravenously. At 24 h after the endotoxin challenge, lungs, livers and striated muscle of skin were analyzed for their myeloperoxidase activity. To monitor intravital leukocyte-endothelial cell interactions, fluorescence videomicroscopy was performed in the skin fold chamber model of the BALB/c mouse at 3, 8 and 24 h after injection of endotoxin. Results: Myeloperoxidase activity at 24 h after the endotoxin challenge in lungs (12,171 +/- 2,357 mU/g tissue), livers ( 2,204 +/- 238 mU/g) and striated muscle of the skin ( 1,161 +/- 110 mU/g) was significantly reduced in both treatment groups as compared to controls, with strongest attenuation in the PECAM-1 IgG treatment group. Arteriolar leukocyte sticking at 3 h after endotoxin (230 +/- 46 cells x mm(-2)) was significantly reduced in both treatment groups. Leukocyte sticking in postcapillary venules at 8 h after endotoxin ( 343 +/- 69 cells/mm(2)) was found reduced only in the VCAM-1-mAb-treated animals ( 215 +/- 53 cells/mm(2)), while it was enhanced in animals treated with PECAM-1 IgG ( 572 +/- 126 cells/mm(2)). Conclusion: These data show that both PECAM-1 and VCAM-1 are involved in endotoxin-induced leukocyte sequestration in the lung, liver and muscle, presumably through interference with arteriolar and/or venular leukocyte sticking. Copyright (C) 2004 S. Karger AG, Basel

Developing cardiac and skeletal muscle share fast-skeletal myosin heavy chain and cardiac troponin-I expression

Skeletal muscle derived stem cells (MDSCs) transplanted into injured myocardium can differentiate into fast skeletal muscle specific myosin heavy chain (sk-fMHC) and cardiac specific troponin-I (cTn-I) positive cells sustaining recipient myocardial function. We have recently found that MDSCs differentiate into a cardiomyocyte phenotype within a three-dimensional gel bioreactor. It is generally accepted that terminally differentiated myocardium or skeletal muscle only express cTn-I or sk-fMHC, respectively. Studies have shown the presence of non-cardiac muscle proteins in the developing myocardium or cardiac proteins in pathological skeletal muscle. In the current study, we tested the hypothesis that normal developing myocardium and skeletal muscle transiently share both sk-fMHC and cTn-I proteins. Immunohistochemistry, western blot, and RT-PCR analyses were carried out in embryonic day 13 (ED13) and 20 (ED20), neonatal day 0 (ND0) and 4 (ND4), postnatal day 10 (PND10), and 8 week-old adult female Lewis rat ventricular myocardium and gastrocnemius muscle. Confocal laser microscopy revealed that sk-fMHC was expressed as a typical striated muscle pattern within ED13 ventricular myocardium, and the striated sk-fMHC expression was lost by ND4 and became negative in adult myocardium. cTn-I was not expressed as a typical striated muscle pattern throughout the myocardium until PND10. Western blot and RT-PCR analyses revealed that gene and protein expression patterns of cardiac and skeletal muscle transcription factors and sk-fMHC within ventricular myocardium and skeletal muscle were similar at ED20, and the expression patterns became cardiac or skeletal muscle specific during postnatal development. These findings provide new insight into cardiac muscle development and highlight previously unknown common developmental features of cardiac and skeletal muscle. © 2012 Clause et al

Direct Regulation of Striated Muscle Myosins by Nitric Oxide and Endogenous Nitrosothiols

, both through activation of guanylyl cyclase and through modification of cysteines in proteins to yield S-nitrosothiols. While NO affects the contractile apparatus directly, the identities of the target myofibrillar proteins remain unknown. Here we report that nitrogen oxides directly regulate striated muscle myosins..These data show that nitrosylation signaling acts as a molecular “gear shift” for myosin—an altogether novel mechanism by which striated muscle and cellular biomechanics may be regulated

The histology of voluntary muscle in cattle and changes which occur during fattening

Abstract derived from the Introduction section of the thesis: The present investigation does not enter into a general consideration of the histology or voluntary striated muscle but is confined to a few phases of the subject. The occurrence of fat in the muscle fiber will be considered in some detail. Some observations bearing on the changes in striated muscle incident to fattening, growth, and inanition will be presented

Optical properties of tissue measured using terahertz pulsed imaging.

The first demonstrations of terahertz imaging in biomedicine were made several years ago, but few data are available on the optical properties of human tissue at terahertz frequencies. A catalogue of these properties has been established to estimate variability and determine the practicality of proposed medical applications in terms of penetration depth, image contrast and reflection at boundaries. A pulsed terahertz imaging system with a useful bandwidth 0.5-2.5 THz was used. Local ethical committee approval was obtained. Transmission measurements were made through tissue slices of thickness 0.08 to 1 mm, including tooth enamel and dentine, cortical bone, skin, adipose tissue and striated muscle. The mean and standard deviation for refractive index and linear attenuation coefficient, both broadband and as a function of frequency, were calculated. The measurements were used in simple models of the transmission, reflection and propagation of terahertz radiation in potential medical applications. Refractive indices ranged from 1.5 ± 0.5 for adipose tissue to 3.06 ± 0.09 for tooth enamel. Significant differences (P<0.05) were found between the broadband refractive indices of a number of tissues. Terahertz radiation is strongly absorbed in tissue so reflection imaging, which has lower penetration requirements than transmission, shows promise for dental or dermatological applications