(means 7.30 and 4.89 mmHg respectively; p=0,00), lower bladder ca- pacity (means 0.33 and 0.71 ml respectively; p=0,00) and lower compliance (means 0.03 ...
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MP8-04 UNDERACTIVE BLADDER IN OBESE-PRONE RATS FED A HIGH FAT DIET Nazema Siddiqui*, Alexis Dieter, Cindy Amundsen, Jillene Brooks, Danielle Degoski, Matthew Fraser, Durham, NC
Source of Funding: Ann & Robert H. Lurie Children’s Hospital of Chicago
MP8-03 SKELETAL MYOGENIC DIFFERENTIATION OF URINE-DERIVED STEM CELLS, ANGIOGENESIS AND INNERVATION USING HYDROGEL LOADED WITH GROWTH FACTORS FOR POTENTIAL IN TREATMENT OF URINARY INCONTINENCE Guihua Liu, Rajesh Pareta, Rongpei Wu, Yingai Shi, Chunhua Deng, Xiaobo Zhou, Anthony Atala, Emmanuel Opara, Yuanyuan Zhang*, Winston Salem, NC 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), n¼ 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. Source of Funding: None
INTRODUCTION AND OBJECTIVES: Changes in systemic metabolism lead to alterations in lower urinary tract (LUT) physiology. High fat diets (HFD) are implicated in the development of insulin resistance. Obese Prone (OP) and Obese Resistant (OR) rats (Charles River) are used in obesity research, as OP rats become obese on HFD while OR remain lean. To assess the effect of obesity on LUT function, we characterized LUT function in both rat strains during chronic HFD feeding. METHODS: Four OP and 4 OR female rats were placed on the same HFD at 9 weeks (wks) of age (30% fat for 12 wk, followed by 60% fat for 4 wk, Research Diets). Conscious restrained cystometry was performed at 7, 11 and 15 wk HFD. Body mass index (BMI) was calculated, and serum glucose was measured. For all cystometric evaluations, single fill cystometrograms were performed to determine true bladder capacity (TBC), followed by 60 min of continuous cystometry to determine functional bladder capacity (FBC). Voiding efficiency (%VE) was calculated as follows: %VE ¼ [average FBC/TBC] *100. For animals showing overflow incontinence (OI), TBC was a priori considered to be 5 milliliters. RESULTS: Mean BMIs were higher in OP compared to OR rats (0.64 vs. 0.46 at 7wk, 0.72 vs. 0.52 at 11 wk, 0.74 vs. 0.52 at 15 wk, respectively; p ¼ 0.002). At 15 wk HFD, there were no significant differences in mean blood glucose between OP and OR groups (102 þ/-14 vs. 97 þ/- 11, respectively; p¼0.58). TBC and %VE were compared across all time points and results are shown in the figure. At 7 wk HFD, all rats exhibited normal voiding patterns. At 11 wk HFD, 2/4 (50%) of OP rats exhibited OI while the remaining 2/4 OP and all 4 OR rats exhibited normal voiding patterns. At 15 wk HFD, one of the OP rats with OI at 11 wk HFD could no longer be assessed due to catheter issues. Of the remaining OP rats, 2/3 (66%) exhibited overflow incontinence while all OR rats showed normal voiding patterns. Mean %VE decreased in the OP group. CONCLUSIONS: Obese non-diabetic animals exhibited elevated bladder capacities, to the point of OI in the majority, and decreased voiding efficiency after 15 wk HFD. OP rats given a chronic HFD may serve as a promising new model to study underactive bladder. Source of Funding: Charles B Hammond Research Award, Duke University; Dr. Siddiqui is supported by award number K12-DK100024 from the National Institute of Diabetes and Digestive and Kidney Diseases. Dr. Fraser is supported by grants DoD SCIRP IIReSC110031, VA RRD RX-000876-01A1, and R01 DK 57284.
MP8-05 OBESITY-INDUCED METABOLIC SYNDROME PROMOTE LOWER URINARY TRACT SYMPTOMS (LUTS) IN A MOUSE MODEL Qiqi He, Sanjeev Shukla, Cleveland, OH; Zhiping Wang, Lanzhou, China, People’s Republic of; Guiming Liu, Bernadette Erokwu, Chris Flask, Lan Lu, Melissa Babcook, Firouz Daneshgari, Sanjay Gupta*, Cleveland, OH INTRODUCTION AND OBJECTIVES: Obesity is a major risk factor involved in the development of metabolic syndrome (MS). Accumulating evidences suggests that obesity and MS contributes to lower urinary tract symptoms (LUTS) through alterations in the phenotype of bladder and prostate. The relationship between MS and LUTS has recently been evaluated in humans; however, there is lack of appropriate in vivo models to allow further exploration of those associations. Animal models may provide new insights into the molecular mechanisms of obesity-mediated LUTS to facilitate diagnosis and therapeutic interventions. METHODS: Mice on C57BL/6N and B6.V-Lepob/J background were obtained from the Jackson Laboratory at 6 weeks of age and
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maintained on regular diet for 30 weeks. Animals were subjected to the assessment of body weight (BW), body length (BL), waist circumference (WC), body mass index (BMI), blood glucose (BG), plasma insulin (INS), plasma leptin (LEP), total cholesterol (CHO), free fatty acid (FFA) and evaluated for urinary voiding functions. Total body fat measurement, prostate and bladder volumes were analyzed by MRI followed by histological evaluation of the organs. These parameters were used to examine the associations between MS and LUTS. RESULTS: Obesity parameters such as BW, WC, and BMI were significantly higher in B6.V-Lepob/J mice compared to C57BL/6N mice (p
