Utilizing techniques developed in our laboratory, we propose to mount a systematic in vitro and in vivo physiological and biomechanical analysis of muscle stem cell based tissue engineering treatment of stress urinary incontinence (SUI). SUI is a significant medical problem affecting approximately 25 million American women 1. Despite the high prevalence of SUI, there is very little treatment-oriented research utilizing tissue engineering techniques. Using such techniques, we propose to develop a truly physiologic sling, not from synthetic or cadaveric tissue, but rather an engineered, functional stem cell muscle scaffold that can be implanted to repair a damaged urethral sphincter.Hypothesis: 1) Muscle Derived Stem Cells (MDSC) improve the mechanical properties of small intestinal submucosa (SIS) scaffold in vitro. 2) MDSC/SIS suburethral sling improves continence in an in vivo rat model of SUI.The Key Aims of this grant include: 1) Establish the mechanisms underlying the alterations in biaxial mechanical properties of MDSC-seeded SIS: a) Establish the feasibility of seeding MDSC to SIS; b) Establish the long-term persistence of MDSC on SIS; and c) Evaluate the compliance properties of SIS alone and MDSC/SIS in vitro and in vivo with comprehensive mechanical testing at different time points. 2) Evaluate the contractile properties of MDSC/SIS: a) Test muscle and neurally evoked contractile properties of MDSC/SIS based on seeding different densities of MDSC onto SIS and culturing for 7 days; b) Using the best seeding density from 2a, test muscle and neurally evoked contractility properties of MDSC/SIS, culture in vitro at different time points; and c) Using the best seeding density from 2a, test muscle and neurally evoked contractility properties of MDSC/SIS cultured in vivo and at different time points. 3) Assess the functional outcome of MDSC/SIS versus SIS suburethral sling placement following simulated birth trauma: a) Assess the effect of SIS or MDSC/SIS suburethral slings on micturition during continuous cystometry; b) Assess the effect of SIS or MDSC/SIS on leak point pressure (LPP) using the reliable and sensitive vertical tilt intravesical pressure clamp method; c) Evaluate the immunohistology of implanted SIS and MDSC/SIS suburethral slings; and d) Retest the mechanical properties as well as muscle and neurally evoked contractile properties of MDSC/SIS suburethral slings following LPP testing for correlative purposes. By reengineering the deficient urinary sphincter through functional tissue engineering, we plan to significantly improve the treatment of SUI. We want to strongly emphasize that our research is in complete compliance with the federal guidelines on embryonic stem cell research. These stem cells have not been obtained from embryos (animal or human) or cell lines of embryonic stem cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR049398-04
Application #
7124691
Study Section
Special Emphasis Panel (ZRG1-UROL (01))
Program Officer
Nuckolls, Glen H
Project Start
2003-05-01
Project End
2008-04-30
Budget Start
2006-05-01
Budget End
2008-04-30
Support Year
4
Fiscal Year
2006
Total Cost
$275,829
Indirect Cost
Name
University of Pittsburgh
Department
Urology
Type
Schools of Medicine
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Kitta, Takeya; Yoshikawa, Satoru; Kawamorita, Naoki et al. (2016) The effect of ovariectomy on urethral continence mechanisms during sneeze reflex in middle-aged versus young adult rats. Neurourol Urodyn 35:122-7
Kawamorita, Naoki; Kaiho, Yasuhiro; Miyazato, Minoru et al. (2015) Roles of the spinal glutamatergic pathway activated through ?-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptors and its interactions with spinal noradrenergic and serotonergic pathways in the rat urethral continence mechanisms. Neurourol Urodyn 34:475-81
Sumino, Yasuhiro; Yoshikawa, Satoru; Mimata, Hiromitsu et al. (2014) Therapeutic effects of IGF-1 on stress urinary incontinence in rats with simulated childbirth trauma. J Urol 191:529-38
Yoshikawa, Satoru; Kitta, Takeya; Miyazato, Minoru et al. (2014) Inhibitory role of the spinal cholinergic system in the control of urethral continence reflex during sneezing in rats. Neurourol Urodyn 33:443-8
Koike, Yusuke; Furuta, Akira; Suzuki, Yasuyuki et al. (2013) Pathophysiology of urinary incontinence in murine models. Int J Urol 20:64-71
Yoshimura, Naoki; Miyazato, Minoru (2012) Neurophysiology and therapeutic receptor targets for stress urinary incontinence. Int J Urol 19:524-37
Kitta, Takeya; Haworth-Ward, Donna J; Miyazato, Minoru et al. (2011) Effects of ovariectomy and estrogen replacement on the urethral continence reflex during sneezing in rats. J Urol 186:1517-23
Furuta, Akira; Suzuki, Yasuyuki; Asano, Koji et al. (2011) Urethral compensatory mechanisms to maintain urinary continence after pudendal nerve injury in female rats. Int Urogynecol J 22:963-70
Kitta, Takeya; Miyazato, Minoru; Chancellor, Michael B et al. (2010) Alpha2-adrenoceptor blockade potentiates the effect of duloxetine on sneeze induced urethral continence reflex in rats. J Urol 184:762-8
Smaldone, M C; Chen, M L; Chancellor, M B (2009) Stem cell therapy for urethral sphincter regeneration. Minerva Urol Nefrol 61:27-40

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