VEGFexpressing control. Additionally, the total number of implanted cells expressing human nuclear markers was significantly higher in VEGF-expressing grafts. More cells also expressed endothelial mark- ers (CD 31 and vWF) and smooth muscle markers (-smooth muscle actin, desmin and myosin) in the VEGF group. Finally, the number of peripheral nerve fibers observed in the USC/Ad-VEGF plus endothelial cells group increased compared to the other controls in vivo. However, a few human nuclei staining were observed in the newly-formed nerve fibers, suggesting that most of the regenerated nerve fibers were derived from host nerve tissue. CONCLUSIONS: VEGF over-expression in implanted USC en- hanced the in vivo survival of these cells and increased endothelial and smooth muscle differentiation of USC. Neovascularization and nerve regeneration significantly increased within the USC/Ad-VEGF grafts. This approach might have important clinical implications for urological cell therapy, including the correction of neurovascular ED. Source of Funding: None 758 OXIDATIVE STRESS-INDUCED SYSTEMIC AND CAVERNOSAL MOLECULAR ALTERATIONS – THEIR ROLE IN THE PROGRESSION OF DIABETIC ERECTILE DYSFUNCTION Angela Castela, Pedro Gomes, Pedro Coelho, Pedro Vendeira, Porto, Portugal; Ronald Virag, Paris, France; Carla Costa*, Porto, Portugal INTRODUCTION AND OBJECTIVES: Erectile Dysfunction (ED) is a common complication of diabetes. Increased oxidative stress (OS) contributes to corporeal deterioration, altering diabetic penile tissue homeostasis. However, it remains unclear the effects of OS in diabetic corpus cavernosum (CC) with the progression of the disease and its role in the development of ED. We aimed to evaluate/quantify systemic and cavernosal alterations caused by OS in early and estab- lished diabetes. METHODS: Male Wistar rats were divided into groups (n=5/ group): 2 and 8 weeks of streptozotocin-induced diabetes and age- matched controls. Blood was withdrawn, 24-hour urine was collected and penises were harvested. All samples were processed for the evaluation/quantification of OS-induced injuries using established bio- markers. Systemic OS was assessed on blood samples by the chro- matographic detection of oxidized glutathione (GSSG)/reduced gluta- thione (GSH) and in urine by evaluating the production of hydrogen peroxide (H2O2). Locally, in CC, the noxious effects of OS were analyzed by: quantifying the production of H2O2, evaluating protein structural alterations by immunohistochemistry for 3-nitrotyrosine (3- NT) and by assessing DNA-induced oxidative lesions through 8-hy- droxydeoxyguanosine (8-OHdG) immunodetection. The expression of 3-NT and 8-OHdG was also evaluated in human non-diabetic and diabetic cavernosal samples. Immunohistochemical quantification of these markers was performed using ImageJ. RESULTS: Our results showed that particularly at 8-week of diabetes there was an increase in blood ratio of GSSG/GSH and of urinary H2O2 levels, indicative of systemic OS augmentation. In CC, H2O2 production was only significant in the established group. Con- cordantly, 3-NT and 8-OHdG expression was increased in 8-week diabetic erectile tissue, as well as, in human diabetic CC, indicating that more severe alterations, as protein nitration and DNA nucleoside modification, occur at a later stage of the disease. CONCLUSIONS: Our study suggested that systemic and penile OS effects are detected mostly in established diabetes. Locally, OS- induced penile protein and DNA lesions present in advanced diabetes may be responsible for hampering functional molecular and cellular processes, culminating with the progression of diabetic-associated ED. Source of Funding: Portuguese Foundation for Science and Technology (PTDC/SAU-OSM/65599/2006) 759 ADIPOSE DERIVED STEM CELLS WITH COX1-10AA-PGIS GENE IMPROVE ERECTILE FUNCTION IN RATS WITH BILATERAL CAVERNOUS NERVE CRUSH INJURY Haocheng Lin*, Houston, TX; Jiuhong Yuan, Chengdu, –; Ke-he Ruan, Houston, TX; Yutian Dai, Nanjing, TX; Run Wang, Houston, TX INTRODUCTION AND OBJECTIVES: Erectile dysfunction (ED) is a common complication after radical prostatectomy. COX1- 10aa-PGIS is a newly engineered protein with COX-1 and prostacyclin synthase activities that converts arachidonic acid directly to prostacy- clin (PGI2). PGI2 is a potent smooth muscle relaxant. This study was designed to use Adipose Derived Stem Cells as the carrier of COX1- 10aa-PGIS gene to improve the penile rehabilitation after bilateral cavernous nerve crush (BCNC) in the rat ED model. METHODS: Adipose Derived Stem Cells (ADSCs) were iso- lated from perigonadal fat of Sprague-Dawley(SD) rats and cultured in DMEM/F12 medium. The COX1-10aa-PGIS gene was transferred into ADSCs. BCNC in adult SD rats was used to replicate radical prosta- tectomy induced ED. SD rats were randomly assigned into three groups: 1. sham surgery; 2. BCNC; 3. BCNC + ADSC with COX1- 10aa-PGIS gene intracavernous injection; 28 days later, intracavern- ous pressure (ICP) is recorded under cavernous nerve stimulation; meanwhile the blood pressure is monitored. At the end of the meas- urement, the penis was harvested and processed for Western blot analysis of COX1-10aa-PGIS.The urine was collected for LC-MS/MS of 6-keto-PGF1a ´. RESULTS: 1. The stable cell line with COX1-10aa-PGIS gene which can over-express PGI2 was identified by LC-MS/MS, G418 and Western blot. 2. Western blot and LC-MS/MS show COX1-10aa-PGIS was successfully expressed. 3. ADSCs with COX1-10aa-PGIS gene therapy improved erectile function in rats after BCNC injury. CONCLUSIONS: These results suggested that COX1-10aa- PGIS gene can be successfully transferred into ADSCs for gene therapy for the treatment of ED after cavernous nerve injury in an animal model. Further study will look at the underlying mechanisms of the ADSC with COX1-10aa-PGIS gene as a potential candidate in post radical prostatectomy penile rehabilitation. Source of Funding: NIH Grants HL56712 and HL79389 760 EXPERIMENTAL EVIDENCE THAT A COMBINATION OF SGC STIMULATOR BAY 60-4552 AND PDE5 INHIBITOR VARDENAFIL MIGHT SALVAGE PATIENTS WITH INSUFFICIENT RESPONSE TO PDE5 INHIBITORS AFTER CAVERNOUS NERVE INJURY Alexandra Oudot, Delphine Behr-Roussel, Sarah Poirier, Orsay, France; Peter Sandner, Wuppertal, Germany; Jacques Bernabe ´, Orsay, France; Francois Giuliano*, Garches, France INTRODUCTION AND OBJECTIVES: Radical prostatectomy (RP) is frequently responsible for erectile dysfunction (ED). Post-RP patients often show insufficient response to PDE5 inhibitor based ED-therapy. This study was undertaken to evaluate the acute effects of the soluble guanylate cyclase (sGC) stimulator, BAY 60-4552 and vardenafil administered alone or in combination on erectile responses to electrical stimulation of the cavernous nerve (ES-CN) in rats with cavernous nerve (CN) crush injury-induced ED. METHODS: Male Sprague-Dawley rats underwent laparotomy (sham, n=10) or bilateral CN crush injury (n=57). After 3 weeks of recovery, erectile function was evaluated in urethane-anesthetized rats following ES-CN at different frequencies. The acute effects of intrave- nous (iv) injection of vehicle, vardenafil 0.03 mg/kg, BAY 60-4552 0.03 mg/kg or 0.3 mg/kg, or a BAY 60-4552 0.03 mg/kg + vardenafil 0.03 mg/kg combination were evaluated in CN crushed rats. RESULTS: Bilateral CN crush injury followed by a 3-week recovery period decreased erectile responses to ES-CN by about 50%. In CN crushed rats, both iv vardenafil 0.03 mg/kg and BAY 60-4552 at Vol. 185, No. 4S, Supplement, Monday, May 16, 2011 THE JOURNAL OF UROLOGY e305