Cavernosal Tissue Research Articles

New methods for the monitoring of nocturnal erections.

The golden standard for measuring nocturnal erections is the RigiScan Plus. It is a relatively big and uncomfortable device dating from the previous century. The aim of this perspective is to conceptualize a user-friendly sensor that can be used at home for monitoring nocturnal erections. A literary search is carried out to explore the physiological changes during nocturnal tumescence and detumescence that can be measured non-invasively. Five sensor concepts are considered: plethysmography for penile arterial pulse, displacement sensor for axial length, strain gauges for radial rigidity and circumference, temperature sensors for measuring skin and cavernosal temperature, and a saturation sensor to measure hypoxia in cavernosal tissue during maximal rigidity. We think that due to practical issues, measuring penile length during sleep is impossible. Further research is recommended to investigate the remaining sensor concepts. Whether a combination of these techniques is favorable or only one of them should be studied more thoroughly.

Characterization of the Renin-Angiotensin System in Aged Cavernosal Tissue and its Role in Penile Fibrosis

BackgroundThe renin-angiotensin system (RAS) plays an important role in erectile function. The RAS contains 2 major axes: one deleterious, composed of ACE-Ang II-AT1 receptor, and another protective, composed of ACE2-Ang-(1-7)-Mas receptor. While aging is a well-known cause for development of male sexual disorders, little is known about local regulation of the RAS in age-related erectile dysfunction (ED). AimThe present study aimed to assess regulation of the RAS in aging-associated ED rat model and evaluate possible options for disease management through pharmacological modulation of the RAS. MethodsPenile tissues were harvested from 3-, 12-, and 24-month-old Wistar rats. Local expression of major RAS components and ED markers was measured by RT-PCR. Protein expression of RAS components was assessed by western blot. Collagen deposition was measured by Sirius Red and immunohistochemical staining. Evaluation of collagen content was also performed in penile sections of Mas-knockout mice by Sirius Red and Masson’s trichrome stainings. Finally, the effect of Ang-(1-7) pretreatment on TGF-β–induced myofibroblast activation was studied in primary cavernosal and immortalized fibroblasts. OutcomesExperimental results highlighted the essential role of the RAS in modulation of cavernosal fibrosis. ResultsThe present study demonstrates local expression of angiotensinogen mRNA alongside with major RAS components, which suggests local autonomous functioning of the RAS within penile tissue. Gene expression analysis revealed strong positive correlation between ACE-Ang II-AT1 axis with markers for inflammation and fibrosis. While corpus cavernosum from 24-month-old rats was characterized by increased collagen deposition, protein expression of ACE, AT1, and Mas was shown to be upregulated in the penile tissue of this group. At the same time, penile sections from Mas-knockout mice (FVB/N background) were also shown to have increased collagen deposition. Finally, it was demonstrated that Ang-(1-7) treatment of primary cavernosal and immortalized fibroblasts was able to alleviate TGF-β–induced fibroblast-to-myofibroblast transition. Clinical TranslationThe present study suggests Ang-(1-7) treatment as a possible strategy for pharmacological management of fibrosis-associated ED in aging. Strengths & LimitationsThe link between the RAS and penile fibrosis, indicated by a holistic screening of different ED markers, was confirmed by in vivo and in vitro data. However, results, presented in the manuscript, need to be further reinforced by human data. Important to note, the main goal of the study was to characterize RAS regulation in aging condition rather than state any causal relationships. ConclusionPresent study characterizes RAS regulation in aging-associated ED and indicates its important role in cavernosal fibrosis.Bragina ME, Costa-Fraga F, Sturny M, et al. Characterization of the Renin-Angiotensin System in Aged Cavernosal Tissue and its Role in Penile Fibrosis. J Sex Med 2020;17:2129–2140.

Thiol/Disulfide Homeostasis in Patients With Erectile Dysfunction

BackgroundAlthough there are no sufficient data on association between oxidative stress and erectile dysfunction (ED), numerous studies have reported that imbalance between the formation of reactive oxygen species and body's antioxidant defenses may play a role in the pathogenesis of ED. AimThe aim of this study was to determine and compare the oxidant and antioxidant status in patients with ED and healthy controls with a novel automated assay for thiol/disulphide homeostasis test. MethodsOur study included 123 patients with ED and 90 healthy individuals. ED was evaluated by asking questions 1–5 and 15 of the International Index of Erectile Function form. In this study, we used Erel and Neselioglu’s thiol/disulfide homeostasis test, which is one of the novel methods that can measure both variables of the oxidative/antioxidative balance individually and collectively. OutcomesThis method measured serum antioxidant (total thiol [toSH], native thiol [SH]) and oxidant (disulfide [SS]) levels. The statistical comparisons were performed between patients with ED (ED+ group) and without ED (ED− group) first and then within the ED+ group. After toSH, SH, and SS levels were determined; SS/toSH%, SS/SH%, and SH/toSH% levels were analyzed separately and compared statistically. ResultsWe found a significant difference between ED− and ED+ groups in terms of toSH, SH, SS/toSH%, and SS/SH% ratios. SS parameters were increased in patients with ED, but there was no significant difference in terms of SS and SH/toSH% values. Clinical ImplicationsClarification of the factors involved in the etiology of ED such as oxidative/antioxidative balance may open new grounds in the early diagnosis and treatment of the disease. Strengths & LimitationsIt is a prospective, randomized clinical study with the use of a novel, reliable, and fully automated technique. The limitations of the study are use of a subjective tool such as the International Index of Erectile Function, obtaining blood samples from the peripheral vein instead of penile cavernosal tissue, and relatively small sample size. ConclusionThe results of this study showed that thiol/disulfide homeostasis is altered in ED, and this imbalance may be a factor in its pathophysiology. We determined that as ED gets more severe, toSH and SH parameters decrease, whereas SS parameter increases.Micoogullari U, Karatas OF, Kisa E, et al. Thiol/Disulfide Homeostasis in Patients With Erectile Dysfunction. J Sex Med 2020;17:1934–1941.

Erectile dysfunction is associated with defective L-cysteine/hydrogen sulfide pathway in human corpus cavernosum and penile arteries

H2S signaling was proposed to participate in erectile physiology. L-cysteine (CYS)/H2S pathway stimulation causes cGMP-dependent relaxation of human corpus cavernosum (HCC) and penile arteries (HPRA). The aim was to evaluate the impact of ED on CYS/H2S pathway at functional and molecular level in human penile vascular tissues. NaHS- and CYS-induced responses were evaluated in HCC and HPRA from organ donors without ED (NoED, n = 29) and from ED patients undergoing penile prosthesis insertion (n = 45). cGMP accumulation and cystathionine β-synthase and cystathionine γ-lyase expression were also determined.NaHS-induced relaxations were slightly but significantly impaired in HCC but not in HPRA from ED patients. In contrast, CYS-induced relaxations were markedly impaired in HCC (Emax 67.6 ± 4.9% vs 46.2 ± 4.6%, P < 0.01) and HPRA (Emax 80.8 ± 4.0% vs 48.1 ± 8.6%, P < 0.05) from men with ED. Impairment of CYS-induced responses was observed even after separating diabetic ED patients. In HPRA from ED patients, CYS- but not NaHS-induced vasodilation was significantly associated to endothelial function measured as vasodilatory capacity of acetylcholine (ACh) in these preparations (r2 = 0.481, P < 0.01). Impairment of CYS-induced relaxations was related to significant reduction in CYS-induced accumulation of cGMP in cavernosal tissue. Furthermore, the expression of H2S synthesizing enzymes was significantly reduced in HCC from ED patients with respect to NoED. This was confirmed by immunofluorescence in HCC and HPRA sections. ED involves impairment of CYS/H2S pathway in penile vascular tissues associated with decreased expression of H2S generating enzymes, CBS and CSE. These evidences support a therapeutic potential for modulation of CYS/H2S signaling in the management of ED.

Melatonin prevents deterioration of erectile function in streptozotocin-induced diabetic rats via sirtuin-1 expression.

A review of the literature indicated that sirtuin-1 expression, a regulator of nitric oxide bioavailability in erectile dysfunction (ED) after melatonin therapy, has not yet been investigated. The objective of this study was to evaluate the protective effects of melatonin for erectile function with sirtuin-1 protein expression in type 1 diabetic rat models. Fifty male Sprague Dawley rats were placed into five groups. Except for those in the control group (C), each animal received a single dose (60mg/kg) of streptozotocin to induce diabetes. The animals were placed into the diabetes (D) group, insulin (I) group (6 U/kg/day), melatonin (Mel) group (10mgkg-1 day-1 ) and combined treatment (I+Mel) group. Ten weeks later, the serum testosterone levels, intracavernosal pressure (ICP), mean arterial pressure (MAP), malondialdehyde (MDA), cyclic guanosine monophosphate (c-GMP), 8-hydroxydeoxyguanosine (8-OHdG), nitric oxide synthase (NOS), caspase-3 activity, sirtuin-1 and endothelial nitric oxide synthase (eNOS) protein expression and histological findings were assessed. The mean ICP/MAP ratio for the D group was lower than the mean ratios for the other groups. The treatment groups, particularly the I+Mel group, exhibited lower 8-OHdG and MDA levels and caspase-3 activity than the D group. The sirtuin-1 and eNOS expression and cavernosal tissue (CT) histology seemed to have been preserved by the melatonin and/or insulin therapy. These results were indicative of a profound protective effect of melatonin by the activation of sirtuin-1 protein expression against hyperglycemia-induced oxidative CT injury.

The protective effect of Ferula elaeochytris on age-related erectile dysfunction

Ethnopharmacological relevanceFerula elaeochytris Korovin (FE) is a perennial medicinal plant of Apiaceae family. Ferula elaeochytris Korovin, known as ‘Çakşır’ in Anatolia, is widely used as an aphrodisiac as well as antioxidant, anti-inflammatory, and anti-diabetic.Aim of the study: Erectile dysfunction (ED) is a serious public health problem that has a high prevalence and negatively affects the quality of life in elderly men. In the treatment and prophylaxis of many diseases, because of widely increasing use of plant extracts as therapeutic agents, preclinical studies related to plant extracts are becoming more important by the day. In this study, we aimed to investigate the protective effect of Ferula elaeochytris Korovin (FE) root extract on age-related ED. Materials and methodsSeventy-two male Wistar albino rats were equally divided into four groups: 4-month aged rats (Y), 24-month aged rats (AG), and FE-administered (20 and 40 mg/kg/day; oral gavage; over 8 weeks) 24-month aged rats (AG + FE). The measurements included: changes in smooth muscle cells and collagen fibrils, tumor necrosis factor alpha (TNF-α), penile neuronal nitric oxide synthase (nNOS) and endothelial nitric oxide synthase (eNOS) expression, serum testosterone concentrations (ST), neurogenic- and endothelial-dependent relaxations of the corpus cavernosum (CC), intracavernosal pressure/mean arterial pressure (ICP/MAP), area under the curve (total ICP), total antioxidant status (TAS), and total oxidant status (TOS) on corpus cavernosal tissue. ResultsThese results have an important role in the development of ED. ICP/MAP, total ICP, eNOS/nNOS expressions and ST levels increased in AG+40 mg FE group compared to the AG group, whereas TNF-α levels decreased and oxidative and antioxidant parameters balanced. ConclusionsOur findings show that FE may have a useful effect on decelerating the development of age-related ED.

MP84-09 DEFECTIVE L-CYSTEINE/HYDROGEN SULFIDE PATHWAY IN HUMAN CORPUS CAVERNOSUM AND PENILE ARTERIES FROM MEN WITH ERECTILE DYSFUNCTION

You have accessJournal of UrologySexual Function/Dysfunction: Basic Research & Pathophysiology (MP84)1 Apr 2020MP84-09 DEFECTIVE L-CYSTEINE/HYDROGEN SULFIDE PATHWAY IN HUMAN CORPUS CAVERNOSUM AND PENILE ARTERIES FROM MEN WITH ERECTILE DYSFUNCTION Alejandro Sevilleja-Ortiz, José M. La Fuente, Juan Ignacio Martínez-Salamanca*, Esaú Fernández-Pascual, Joaquín Carballido, and Javier Angulo Alejandro Sevilleja-OrtizAlejandro Sevilleja-Ortiz More articles by this author , José M. La FuenteJosé M. La Fuente More articles by this author , Juan Ignacio Martínez-Salamanca*Juan Ignacio Martínez-Salamanca* More articles by this author , Esaú Fernández-PascualEsaú Fernández-Pascual More articles by this author , Joaquín CarballidoJoaquín Carballido More articles by this author , and Javier AnguloJavier Angulo More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000000976.09AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Hydrogen sulfide (H2S) signaling has been proposed to participate in erectile physiology. L-cysteine (CYS)/H2S pathway stimulation causes cGMP-dependent relaxation of human corpus cavernosum (HCC) and penile arteries (HPRA). The aim of this work was to evaluate the impact of erectile dysfunction (ED) on CYS/H2S pathway at functional and molecular level in human penile vascular tissues. METHODS: NaHS- and CYS-induced responses were evaluated in HCC and HPRA from organ donors without notice of ED (NoED, n=29) and from ED patients undergoing penile prosthesis insertion (n=45). cGMP accumulation and cystathionine β-synthase (CBS) and cystathionine γ-lyase (CSE) expressions were also determined in these tissues. RESULTS: NaHS-induced relaxations were slightly but significantly impaired in HCC (pEC50 4.58±0.09 vs. 4.19±0.07, p<0.01) but not in HPRA (pEC50 4.16±0.10 vs. 4.00±0.07) from ED patients. In contrast, CYS-induced relaxations were markedly impaired in both HCC (Emax 67.6±4.9% vs 46.2±4.6%, p<0.01) and HPRA (Emax 80.8±4.0% vs 48.1±8.6%, p<0.05) from men with ED. Impairment of CYS-induced responses was observed even after separating diabetic ED patients. In HPRA from ED patients, CYS- but not NaHS-induced vasodilation was significantly associated to endothelial function measured as vasodilatory capacity of acetylcholine (ACh) in these preparations (pEC25 CYS vs. Emax ACh: r2=0.481, p<0.01). Impairment of CYS-induced relaxations was related to significant reduction in CYS-induced accumulation of cGMP in cavernosal tissue. Furthermore, the expression of H2S synthesizing enzymes CBS and CSE, as determined by Western blot, was significantly reduced in HCC from ED patients with respect to NoED tissues. This diminished expression of CBS and CSE was clearly manifested in immunofluorescence assays in HCC and HPRA sections. CONCLUSIONS: ED involves impairment of CYS/H2S pathway in penile vascular tissues associated with decreased expression of H2S generating enzymes, CBS and CSE. These evidences support a therapeutic potential for modulation of CYS/H2S signaling in the management of ED. Source of Funding: None © 2020 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 203Issue Supplement 4April 2020Page: e1268-e1268 Advertisement Copyright & Permissions© 2020 by American Urological Association Education and Research, Inc.MetricsAuthor Information Alejandro Sevilleja-Ortiz More articles by this author José M. La Fuente More articles by this author Juan Ignacio Martínez-Salamanca* More articles by this author Esaú Fernández-Pascual More articles by this author Joaquín Carballido More articles by this author Javier Angulo More articles by this author Expand All Advertisement PDF downloadLoading ...

Evaluation of effectiveness of antiandrogen treatments on cavernosal tissues in rats.

In this study, we aimed to compare changes in cavernosal tissues in rats with antiandrogen treatment and orchiectomy. A total of 42 Wistar albino rats were divided into four groups. Group I, control group,Group II, LH-RH was given for 1month, Group III-LH-RH+Bicalutamide was given for 1month, and Group IV was defined as orchiectomy and followed up for 1month. Measurements of intracavernosal pressure with different electrical stimuli and pathological findings of smooth muscle collagen in cavernosal tissues were examined. While the cavernosal pressure response in all the different electrical stimuli given in the control group and in all other groups was significantly lower than that in the other groups, it was statistically significant at 7.5 and 10V (p=.005, p<0001). According to the pathologic evaluation, the density of tissue collagen increased significantly in the other groups according to the control group. In groups 3 and 4, the density of 4+ collagen was found to be increased according to Groups 1 and 2. In the LH-RH alone group, it appears that there are no 4+ colloid density and less damage. According to these findings, the negative effect of LH-RH treatment on cavernosal tissues appears to be less.

Enhanced Contribution of Orai Channels to Contractility of Human Penile Smooth Muscle in Erectile Dysfunction

BackgroundStore-operated calcium entry and its key players, stromal interaction molecule (STIM) and Orai calcium channels, have been proposed as emergent therapeutic targets in cardiovascular pathophysiology. We hypothesize alteration of STIM/Orai signaling in erectile dysfunction (ED). AimTo evaluate the contribution of STIM/Orai to human penile tissue contraction and to analyze the influence of ED on STIM/Orai signaling at functional and expression levels in human penile vascular tissues. MethodsHuman penile resistance arteries (HPRA) and human corpus cavernosum (HCC) were dissected from cavernosal specimens from 30 organ donors without history of ED (No ED) and from 48 patients with ED undergoing penile prosthesis insertion and functionally evaluated in wire myographs and organ chambers, respectively. Expression of STIM-1, Orai1, and Orai3 in HCC was localized and quantified by immunofluorescence. Main Outcome MeasuresThe main outcome measures are functional responses in HCC and HPRA and STIM/Orai channel protein expression in human cavernosal tissue. ResultsInhibition of Orai channels with YM-58483 (20 μM) significantly reduced norepinephrine–induced contractions in both HCC and HPRA from either No ED or ED subjects, but the effects were more marked in ED (−20.1 ± 5.9% vs −45.5 ± 13.2% and −15.9 ± 4.0% vs −31.4 ± 6.9% reduction in Emax to norepinephrine in HCC and HPRA, respectively). Thromboxane-induced contractions were reduced and neurogenic contractile and relaxant responses modulated by Orai inhibition in penile tissues from patients with ED. In fact, addition of YM-58483 concentration dependently relaxed precontracted HPRA and HCC. These relaxations were significantly more pronounced in tissues from patients with ED (EC50 7.5 vs 1.3 μM and 10.5 vs 1.3 μM, for HCC and HPRA, respectively). All HCC specimens displayed expression of STIM-1, Orai1, and Orai3. Significantly increased expression of Orai1 and Orai3 but not STIM-1 was observed in patients with ED. Clinical TranslationInhibition of enhanced Orai activity in human penile vascular tissue could facilitate erectile responses, alleviating ED. Strengths and LimitationsEnhanced STIM/Orai activity contribution to penile smooth muscle tone in ED is demonstrated at functional and structural levels in human tissues from a representative sample of patients with ED and in comparison with healthy tissue. We cannot differentiate the specific contribution of risk factors associated with ED to hyperactivity of the Orai system. ConclusionsOrai channels significantly contribute to human penile smooth muscle contraction. Orai contribution to penile smooth muscle tone is functionally enhanced in ED accompanied by increased expression of Orai channels in cavernosal tissue. Orai inhibition could be a potential therapeutic strategy to reduce penile smooth muscle contraction in ED.Sevilleja-Ortiz A, El Assar M, García-Rojo E, et al. Enhanced Contribution of Orai Channels to Contractility of Human Penile Smooth Muscle in Erectile Dysfunction. J Sex Med 2020;17:881–891.

172 Caspase Signalling in ED Patients and Animal Models

Erectile dysfunction (ED) affects ∼50% of men aged 40-70 and has a high impact on men’s health. Current treatments are ineffective in the difficult to treat prostatectomy (16-82%) and diabetic (56-59%) patients due to injury to the cavernous nerve (CN). With denervation the critical smooth muscle (SM) undergoes apoptosis and the penis becomes fibrotic, thus altering the corpora cavernosal architecture. In order to devise novel ED therapies, prevention of corpora cavernosal remodeling is critical. Apoptosis can take place via the intrinsic (caspase 9) or extrinsic (caspase 8) pathway. We examine the mechanism of how apoptosis occurs in ED patients and in a CN injury rat model, to determine points of apoptosis intervention for therapy development. Immunohistochemical analysis and western analysis for caspase 3 cleaved, -8 and -9 (pro and active forms) were performed in corpora cavernosal tissue from Peyronie’s, prostatectomy and diabetic ED patients (n=56), and in penis from adult Sprague Dawley rats that underwent CN crush and were sacrificed after 1-9 days (n=16).

Penile ring entrapment - A true urologic emergency: Grading, approach, and management.

Penile rings are used to sustain erection in order to enhance sexual pleasure. They work by reducing the outflow of blood from the cavernosal tissue. However, if left for extended periods of time severe edema, urethral fistula, gangrene, and even complete loss of the distal penis can ensue, this is known as penile ring entrapment (PRE). Management poses particular challenges due to its rarity. Herein, we report our experience with three patients from our institution that presented with PRE and include a review of the approaches others have taken. We also propose a simpler and more effective grading scale to allow for easier communication between providers, as the current grading scales do not do so.

Non-Invasive Cell Tracking with Brighter and Red-Transferred Luciferase for Potential Application in Stem Cell Therapy

This study investigated the safety of a novel cell-labeling technology with mKATE and Renilla reniformis luciferase (mKATE-renLUC) and assessed the efficacy on tracking implanted human placental stromal cells (PSC) in an erectile dysfunction (ED) animal model. Human PSC were labeled with mKATE-renLUC using a lentivirus. Cell viability, apoptosis, proliferation, migration, surface marker expression and differentiation potential of the labeled PSC were evaluated and compared with non-labeled PSC. The paracrine profile of labeled cells was examined using an angiogenesis protein array. The brightness and duration of labeled cells with different densities were evaluated. An ED rat model was established and labeled PSC were injected into cavernosal tissue of the penis. The migration and distribution of transplanted PSC were monitored using an IVIS imaging system in real time. Implanted PSC were identified in isolated tissues via detection of mKATE fluorescence. The cell viability, morphology, proliferation, migration, surface marker expression and differentiation potential of mKATE-renLUC-labeled PSC were similar to those of non-labeled cells in vitro (no statistical difference p>0.05). Similar expressions of trophic factors were found between labeled and non-labeled PSC. The migration and distribution of PSC expressing renLUC were tracked in vivo using IVIS imaging system. mKATE-positive PSC were detected in penile, kidney, prostate and hepatic tissues using histological methods. This labeling technology provides a safe and effective cell-tracking approach with a brighter fluorophore and codon-optimized luciferase.

Extracorporeal Shock Waves Therapy Delivered by Aries Improves Erectile Dysfunction in Spontaneously Hypertensive Rats Through Penile Tissue Remodeling and Neovascularization.

BackgroundLow-intensity extracorporeal shock wave therapy (Li-ESWT) has been reported to improve erectile function in patients with moderate-to-severe erectile dysfunction (ED) or even convert phosphodiesterase type 5 inhibitors nonresponders to responders. ED is highly prevalent in hypertensive patients. The effect of Li-ESWT on an animal model of hypertension-associated ED has not been reported.AimTo investigate the effect of Li-ESWT on hypertension-associated ED and provide plausible mechanisms of action of Li-ESWT on local mechanisms of penile erection.MethodsSpontaneously hypertensive rats (SHRs) in the active group (n = 13) received Li-ESWT at energy flux density 0.06 mJ/mm2 (Aries; Dornier MedTech, Wessling, Germany) twice weekly for 6 weeks. The emitter was set to zero for SHRs in the sham group (n = 12). Erectile function was assessed 4 weeks post-treatment by monitoring intracavernosal pressure (ICP) in response to electrical stimulation of cavernous nerve before and after single dose of 0.3 mg/kg intravenous sildenafil. Cavernosal tissue was then evaluated for collagen/smooth muscle content, neuronal nitric oxide synthase (nNOS), and vascular endothelial factor (CD31) expression.OutcomesErectile function was assessed with ICP, erectile tissue remodeling was studied by smooth muscle/collagen ratio, nNOS and CD31 were semiquantitatively evaluated on cavernosal sections.ResultsThe improvement of ICP parameters was greater in Li-ESWT–treated rats compared with controls with and without sildenafil. Sildenafil led to 20% increase in area under the intracavernosal pressure curve measured during the entire response/mean arterial pressure at 10 Hz in ESWT_SHR + sildenafil compared with ESWT_SHR. The smooth muscle/collagen ratio increased 2.5-fold in Li-ESWT compared with sham. Expression of CD31 tended to be increased whereas nNOS was unchanged.ConclusionsLi-ESWT by Aries may represent an effective noninvasive therapeutic alternative and a relevant add-on therapy to phosphodiesterase type 5 inhibitors for ED in hypertensive patients, and it is suggested that it acts via remodeling of the penile tissue and promoting cavernosal vascularization.Assaly R, Giuliano F, Clement P, et al. Extracorporeal Shock Waves Therapy Delivered by Aries Improves Erectile Dysfunction in Spontaneously Hypertensive Rats Through Penile Tissue Remodeling and Neovascularization. Sex Med 2019;7:441–450

Abstract P124: Nlrp3 Activation by Endothelin-1 Mediates Neutrophil Recruitment in Cavernosal Tissue of DOCA/salt Mice

Introduction: Endothelin-1 (ET-1) induces penile detumescence and acts as an important mediator of chronic inflammation. NACHT, leucine reach repeat (LRR) and PYD domain-containing protein 3 (NLRP3) releases the active form of IL-1β. This cytokine is involved in neutrophil recruitment from the circulation to the tissues. Increasing evidence shows that neutrophil infiltration is involved in the genesis and aggravation of various cardiovascular diseases. Hypothesis: NLRP3 activation by ET-1 mediates neutrophil infiltration in the corpora cavernosa (CC) of DOCA/salt hypertensive mice. Methods: All protocols were approved by the Animals Use Ethics Committee (CEUA 213/2016). Male C57BL/6 (WT) and NLRP3 deficient mice (NLRP3 -/- ) were either induced to DOCA/salt hypertension or SHAM and treated with bosentan or vehicle. Caspase-1 and IL-1β activities were determined in CC. NF-κB, ICAM-1 and VCAM-1 expressions in CC were determined by immunohistochemistry. Flow cytometry was performed to evaluate the infiltration of myeloid cells into CC. Results: DOCA/salt hypertension increased caspase-1 and IL-1β activities in CC. NF-κB, ICAM-1, VCAM-1 and neutrophil infiltration were increased in CC of DOCA/salt mice. The treatment with bosentan or the absence of NLRP3 prevented all these alterations in DOCA/salt mice. Conclusion: ET-1-induced NLRP3 activation is involved in neutrophil recruitment in CC of DOCA/salt hypertensive mice.

Mesenchymal stem cell therapy improves erectile dysfunction in experimental spinal cord injury.

The aim of this study is to investigate the therapeutic potential of adipose-derived mesenchymal stem cell (AD-MSC) from brown adipose tissue on erectile dysfunction (ED) in experimentally induced spinal cord injury in rats. 24 male Wistar rats were divided into 3 groups; control, spinal cord injury (SCI) + vehicle, and SCI + AD-MSC. To induce SCI, a standard weight-drop method that induced a moderate to severe injury (100 g/cm force) at T7-T10, was used. AD-MSC (3 × 105 cells /5 μL) was applied by local transplantation into the region of injury. At the end of four-weeks, rats underwent neurological examinations and then intracavernosal and mean arterial pressures (ICP and MAP) measurements. After decapitation, spinal cord and cavernosal tissue samples were taken to analyze neuronal nitric oxide synthase (n-NOS), proto-oncogene protein c-FOS and nerve growth factor (NGF). Tissues were also examined histologically. Spinal cord injury caused decrease on NGF and n-NOS levels while c-FOS was increased. The ICP/MAP value in vehicle-treated SCI rats was found to be significantly higher than the control group. On the other hand, in SCI + AD-MSC group, all these parameters were reversed back to control levels. AD-MSC therapy may be beneficial against erectile dysfunction in experimentally induced SCI by ameliorating neuronal damage.

The effect of an antifibrotic agent, pirfenidone, on penile erectile function in an experimental rat model of ischemic priapism.

To date, no effective medical approach for the treatment of erectile dysfunction (ED) secondary to ischemic priapism (IP) has been described. The aim of this study was to evaluate the anti-inflammatory, antifibrotic, and antioxidant effects of pirfenidone (PFD) on cavernosal tissue in a rat model of IP. Forty-eight male albino rats aged 8-10 months, with mean weights of 410 ± 18.6 g were randomized into four groups (n = 12 in each group): no IP (group 1); IP for 1 h, followed by intracavernosal pressure (ICP) measurements using electrical cavernous nerve stimulation (CNS) (group 2); IP for 1 h, followed by ICP measurements using electrical CNS 6 weeks later (group 3); and IP for 1 h, oral PFD (30 mg/kg once daily) treatment by oral gavage, followed by ICP measurements using electrical CNS 6 weeks later (group 4). Malondialdehyde (MDA) and reduced glutathione levels were measured spectrophotometrically. In a histological evaluation, cavernosal collagen/smooth muscle ratios were calculated. The intracavernosal pressure values of group 1 were higher than those of groups 2 and 3 (p < 0.05) but similar to those of group 4 (p > 0.05). The mean MDA level was significantly higher in group 3, as compared with that in group 4 (p = 0.004). The mean collagen/smooth muscle ratio in groups 1-4 was 24%, 42%, 65%, and 48%, respectively. Physiological, biochemical, and histopathological evaluations of the PFD effect on cavernosal tissue in a rat model of IP were the strengths and the lack of molecular and immunohistochemical analysis were the limitations of this study. In this study, we examined the effects of PFD on cavernosal tissue in a rat model of IP. We found that PFD reduced cavernosal fibrotic activity and improved erectile function. We conclude that PFD may represent a new treatment option in IP treatment.

Protective effects of krill oil on ischemic reperfusion injury in experimental model of priapism

Aim: The aim of the study is to evaluate the effects of krill oil on priapism induced ischemia-reperfusion injury in priapism rat model. Methods: Total of 24 rats were randomly divided into three groups with eight rats in each group. Group 1 was determined as the control group. Experimental priapism model was constructed in rats in Groups 2 and 3 for 1 hour and then priapism was terminated for 30 minutes to evaluate ischemia-reperfusion injury. The rats in Groups 1 and 2 were given tap water and standard chow pellets. Group 3: The same feeding procedure was applied to the experimental animals but supplemented with krill oil for one month. At the end of the experiment penectomy were performed, and blood samples were withdrawn from inferior vena cava of the rats to determine the levels of protein carbonyl (PC), malondialdehyde (MDA), nitric oxide (NO), glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) in systemic circulation, and cavernosal tissues.Results: Biochemical examination of penile tissues showed that MDA and PC levels in Group 3 were significantly lower than group 2, while GSH-Px activities were significantly increased (P=0.001, P=0.005, P=0.003 respectively). Serum analysis results showed that MDA and NO levels in Group 3 were significantly lower than Group 2 and SOD activities significantly increased (P=0.011, P=0.001, P=0.009, respectively).Conclusion: In this study, protective effect of krill oil against priapism induced ischemia-reperfusion injury in cavernosal tissue was observed based on biochemical evidence.

The Effects of Oral 5-alpha Reductase Inhibitors on Penile Intracavernosal Pressures and Penile Morphology in Rat Model.

Benign prostatic hyperplasia (BPH), and erectile dysfunction (ED) are urological diseases which affect more than 50 % of men older than 50 years of age. It has been reported that 5-alpha-reductase inhibitors (5-ARIs) used in clinical studies for the treatment of BPH caused ED in 0.8-15.8% of the patients. The aim of this study is evaluation of the effects of oral finasteride and dutasteride on penile intracavernosal pressures and penile morphology in a rat model. Thirty Wistar Albino strain male rats were randomized into control (n = 10), finasteride (n = 10), and dutasteride (n = 10) groups. After 8 weeks of treatment erectile responses were evaluated in all rats measuring intracavernosal pressure (ICP) changes during erectile responses to cavernosal nerve electrical stimulation. Serum hormone levels were studied and all rats underwent prostatectomy and penectomy. All tissue samples were examined histomorphologically and a semiquantitative scoring system was used for cavernosal tissue collagen density grading. One-way analysis of variance was used for statistical analysis and P < .05 was accepted as the level of statistical significance. For two group comparisons Tukey HSD test was used as post hoc test of one way analysis of variance. Approximately 50% decrease was seen in mean ICPs in the finasteride and dutasteride groups compared to the control group for all voltages (2.5 V, 5 V. 7.5 V). Mean ICPs for 7.5 V were 62.17 ± 30.89mmHg in control group, 35.27 ± 31.94 in the finasteride, and 36.01 ± 19.20mmHg in the dutasteride group. But regarding ICPs there was no statistically significant difference between the groups (P > .05). The serum testosterone (T) concentrations were higher in treatment groups (P < .001). Serum dihydrotestosterone (DHT), luteinizing hormone (LH) and follicle stimulating hormone (FSH) concentrations were not significantly different between the groups. As a result of histomorphological studies, a statistically significant increase in cavernosal tissue collagen density, and marked atrophic changes in prostatic epithelial tissues were observed in the treatment groups. Although 5-ARIs cause marked atrophic changes in prostatic epithelial tissues, and prominent collagen deposition in penile cavernosal tissues, no significant effect on penile ICPs was seen in this study. The failure to show a statistically significant difference was attributed to higher standard deviations of ICP values. If sample size and duration of the treatment are increased, statistically significant results in ICPs may be reached. The penile morphology evaluation results point to a negative effect of 5-ARIs on erectile function.

PS-04-001 Aging increases contribution of Orai channels to contractile tone in rat and human corpus cavernosum

The contribution of calcium Orai channels to contractile responses in human corpus cavernosum and penile arteries has been proposed, especially in pathological conditions such as erectile dysfunction (ED). The aim of the present work was to evaluate the influence of aging on Orai channels contribution to cavernosal tissue contraction. The effect of the Orai channel inhibitor, YM-58483 (YM) was evaluated on contractions induced by norepinephrine (NE) in corpus cavernosum (CC) from young (3 months-old) and aged (20 months-old) rats. The effects of YM on endothelial relaxation and neurogenic contraction were also evaluated in tissues from aged rats. As well, the influence of YM on NE-induced contractions was determined in human corpus cavernosum (HCC) from organ donors (OD) without history of ED, which were divided into younger than 65 years (43.5±2.8 years, n=16) and older than 65 years (74.2±3.5 years, n=5). The expression of Orai channels in these HCCs was determined by Western blot.

Co-culture of smooth muscle cells and endothelial cells on three-dimensional bioprinted polycaprolactone scaffolds for cavernosal tissue engineering

Purpose In vitro evaluation of polycaprolactone (PCL) scaffolds fabricated by a three-dimensional (3D) printing technique for tissue engineering applications in the corpus cavernosum. Materials and methods PCL scaffolds were fabricated by use of a 3 D bioprinting system. The 3D-printed scaffolds had interconnected structures for cell ingrowth. Human aortic smooth muscle cells (haSMCs) were seeded on the scaffold and cultured for 5 days, and then human umbilical vein endothelial cells (HUVECs) were also added on the scaffolds and co-cultured with haSMCs for up to 7 days. The ability of these scaffolds to support the growth of HUVECs and haSMCs was investigated in vitro. 3 D strand-deposited scaffolds were characterized by scanning electron microscopy (SEM) images and porosity measurement. Results SEM images showed the surface of the PCL scaffolds to be well covered by HUVECs and haSMCs. Immunofluorescent staining of α-flk1 and α-smooth muscle actin on the HUVECs and haSMCs seeded scaffolds confirmed that the cells remained viable and proliferated throughout the time course of the culture. Conclusion 3 D bioprinting of a PCL scaffold is feasible for co-culturing of HUVECs and haSMCs. This was a preliminary study to investigate the possibility of fabrication of tissue-engineered corpus cavernosum.