MP8-05 OBESITY-INDUCED METABOLIC SYNDROME PROMOTE LOWER URINARY TRACT SYMPTOMS (LUTS) IN A MOUSE MODEL

Abstract

INTRODUCTION AND OBJECTIVES: Despite the potential of stem cells in cell-based therapy in treatment of urinary incontinence, major limitations such as cell retention, ingrowth, and trans-differentiation after implantation remain. To provide safe, site-specific delivery and targeted release of growth factors to implanted stem cells, hyaluronic acid hydrogel with growth factors via heparin provides an alternative to correct tissue deficiency. The goal of this study is to determine whether skeletal myogenic, anigogenic, and neurogenic growth factors released from the hydrogel can induce USCs to give rise to a skeletal myogenic lineage, improve revascularization and innervation, and recruit resident cells to participate in sphincter tissue repair for urinary incontinence. METHODS: Human urine-derived stem cells (USCs) were obtained from eight healthy donors. Hyaluronic acid hydrogel with heparin containing an optimized growth factor cocktail, including VEGF, IGF-1, FGF-1, PDGF-BB, HGF, and NGF. Radiolabeled growth factors were loaded separately and used to access in vitro release from the hydrogel with a gamma counter over 4 weeks. USCs were mixed with the hydrogel and subcutaneously injected in vivo for 4 weeks after implantation. Twenty-four athymic mice were divided into 4 groups (G), n1⁄4 6 animals/G: G1, hydrogel alone; G2, growth factors within gel; G3, USCs with gel; G4, growth factors plus USCs within gel. Cell retention, ingrowth, and differentiation of implanted USCs were examined with real time PCR and immunocytochemical staining. RESULTS: The imbedded growth factors were released quickly in the first three days of incubation followed by a steady rate of release for a month. Numbers of the grafted cells expressing human nuclei significantly increased and more cells expressed myogenic (MyoD, myf5, Myosin) and endothelial cell (CD31 and vWF) transcripts and protein markers in growth factors plus USCs group, compared to other groups. In addition, vessel formation and innervation was significantly more in the group of cell and growth factor in combination than those in controls. CONCLUSIONS: Synergistic effect of growth factors released in a controlled manner from hydrogel efficiently improve cell survival, guide USCs to myogenic differentiation, enhance angiogenesis and innervation, and recruit resident cells to participate in tissue regeneration, which is potential alternative for cell therapy in treatment of urinary incontinence.