Functional Tissue Engineering for Stress Incontinence

描述（由申请人提供）：利用我们实验室开发的技术，我们建议对基于肌肉干细胞的组织工程治疗压力性尿失禁（SUI）进行系统的体外和体内生理和生物力学分析。SUI是一个严重的医疗问题，影响着大约2500万美国女性。尽管SUI的发病率很高，但很少有利用组织工程技术进行治疗的研究。使用这些技术，我们建议开发一个真正的生理吊带，而不是从合成或尸体组织，而是一个工程化的，功能性的干细胞肌肉支架，可以植入修复受损的尿道sphincter.Hypothesis：1）肌肉衍生干细胞（MDSC）提高小肠粘膜下层（SIS）支架的体外力学性能。2)MDSC/SIS尿道悬吊术改善了SUI大鼠模型的力学性能，本基金的主要目的包括：1）建立MDSC接种SIS后双轴力学性能改变的机制：a）建立MDSC接种SIS的可行性; B）建立MDSC在SIS上的长期持久性;和c）通过在不同时间点的综合机械测试，在体外和体内评价单独的SIS和MDSC/SIS的顺应性性质。2)评价MDSC/SIS的收缩性能：a）将不同密度的MDSC接种于SIS上，培养7天，测试MDSC/SIS的肌肉和神经诱发收缩性能：B）采用2a中的最佳接种密度，测试MDSC/SIS的肌肉和神经诱发收缩性能，在不同时间点进行体外培养;和c）使用来自2a的最佳接种密度，测试在体内和在不同时间点培养的MDSC/SIS的肌肉和神经诱发的收缩性能。3)评估模拟产伤后MDSC/SIS与SIS尿道下吊带放置的功能结局：a）评估SIS或MDSC/SIS尿道下吊带在连续膀胱测压期间对排尿的影响; B）使用可靠且灵敏的垂直倾斜膀胱内压力钳方法评估SIS或MDSC/SIS对漏尿点压力（LPP）的影响; c）评估植入的SIS和MDSC/SIS尿道下悬带的免疫组织学;以及d）在LPP测试之后，出于相关目的，重新测试MDSC/SIS尿道下悬带的机械特性以及肌肉和神经诱发的收缩特性。通过功能性组织工程重建有缺陷的尿道括约肌，我们计划显著改善SUI的治疗。我们要强调我们的研究完全符合联邦关于胚胎干细胞研究的指导方针。这些干细胞不是从胚胎（动物或人类）或胚胎干细胞的细胞系中获得的。

DESCRIPTION (provided by applicant): 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.