Neurogenic Erectile Dysfunction Research Articles

Poly-L-lactic acid/gelatin electrospun membrane-loaded bone marrow-derived mesenchymal stem cells attenuate erectile dysfunction caused by cavernous nerve injury

Radical prostatectomy (RP) can cause neurogenic erectile dysfunction (ED), which negatively affects the quality of life of patients with prostate cancer. Currently, there is a dearth of effective therapeutic strategies. Although stem cell therapy is promising, direct cell transplantation to injured cavernous nerves is constrained by poor cell colonization. In this study, poly-L-lactic acid (PLLA)/gelatin electrospun membranes (PGEM) were fabricated to load bone marrow-derived mesenchymal stem cells (BM-MSCs) as a patch to be placed on injured nerves to alleviate ED. This study aimed to establish a promising and innovative approach to mitigate neurogenic ED post-RP and lay the foundation for modifying surgical procedures. Electrospinning and molecular biotechnology were performed in vitro and in vivo, respectively. It was observed that PGEM enhanced the performance of BM-MSCs and Schwann cells due to their excellent mechanical properties and biocompatibility. The transplanted PGEM and loaded BM-MSCs synergistically improved bilateral cavernous nerve injury-related ED and the corresponding histopathological changes. Nevertheless, transplantation of BM-MSCs alone has been verified to be ineffective. Overall, PGEM can serve as an ideal carrier to supply a more suitable survival environment for BM-MSCs and Schwann cells, thereby promoting the recovery of injured cavernous nerves and erectile function.

(202) Comparative Study of the Efficacy of Avanafil and Atorvastatin Daily Dose in Management of Vasculogenic Erectile Dysfunction

Abstract Introduction Erectile dysfunction (ED) is defined as the “consistent inability to attain or maintain a penile erection, or both, sufficient for adequate sexual relations”. ED is not only a medical problem but affects the marital relations and patient's self-esteem and causes the quality of life to decrease. which in turn causes the patient to go into depression or develop anxiety. In the past, erectile dysfunction was considered, in most cases, to be a purely psychogenic disorder, but current evidence suggests that more than 80% of cases have an organic aetiology. Causes of organic erectile dysfunction can now be broadly divided into nonendocrine and endocrine. Of the nonendocrine aetiologies, vasculogenic (affecting blood supply) is the most common and can involve arterial inflow disorders and abnormalities of venous outflow (corporeal veno-occlusion). Avanafil is a second-generation phosphodiesterase type 5 inhibitor approved for erectile dysfunction by the FDA on 2012 and by EMA in 2013. With a high selectivity for phosphodiesterase type 5 isoform AND faster onset of action compared with other PDE5 inhibitors, it offers more tolerability and less adverse events. Atorvastatin is 3-hydroxy-methylglutaryl (HMG)-CoA reductase inhibitor act on the rate-limiting step in cholesterol biosynthesis (the conversion of HMG-CoA to mevalonate) and are effective in treating dyslipidemia. statins reverse endothelial dysfunction by decreasing the action of oxidized low-density lipoprotein (LDL) on endothelial cells, resulting in an increase of NO activity. Objective is to evaluate and compare the efficacy of avanafil and atorvastatin daily dose in management of patients with vasculogenic erectile dysfunction. Methods A 60 adult heterosexual males complaining of vasculogenic erectile dysfunction proven by penile duplex were collected from outpatient andrology clinics of Al-Azhar University Hospitals. The patients were divided into two treatment groups: (Group A): 30 patients received daily dose of 100 mg avanafil. (Group B): 30 patients received daily dose of 40 mg atorvastatin, The main outcome measures and the difference in penile duplex ultrasound results and IIEF-EF scores between the two groups are evaluated and compared at the end of the study, Age of patients between 20 and 50 years, established diagnoses of Vasculogenic erectile dysfunction, Normal lipid profile, Normal Total and Free testosterone levels and No contraindications to the study medications were included criteria. However patients who failed to fulfill any of the inclusion criteria, psychogenic or neurogenic erectile dysfunction, dyslipidemia Patients with Renal or Hepatic insufficiency and Patients with advanced cardiovascular diseases such as heart failure and coronary artery disease on antianginal drugs especially nitrates were excluded. Results Our study revealed statistically significant improvement in Penile Doppler ultrasonography parameters (PSV, EDV& RI) and IIEF-EF scores in avanafil group compared to atorvastatin group. Conclusions avanafil 100 mg daily dose leads to significant improvement in men with vasculogenic erectile, while atorvastatin 40 mg caused no significant differences, neither positively nor adversely. In summary, avanafil at dose of 100 mg can be a therapeutic appropriate choice in cases of vasculogenic ED. Disclosure No.

(061) Xenon Facilitates Schwann Cell Demyelination via Inhibition of SPRY2 in Treating Erectile Function After Cavernous Nerve Injury

Abstract Introduction Cavernous nerve injury induced erectile dysfunction (ED) is the most common complication after radical prostatectomy for prostate cancer. Our previous studies found that xenon (Xe) can partially restore the erectile function and cavernous nerve function of rats with neurogenic ED, but the mechanism is unclear. Schwann cells (SC) play an important role in peripheral nerve repair, and after axonal injury, SC undergo demyelination and transform into repair-type SC to promote repair. Objective The purpose of this study is to verify whether Xe promotes the demyelination of Schwann cells and its molecular mechanism. Methods Human SCs were cultured in vitro and given Echo-lipid particles (ELIPs) carrying Xe. The control group was given empty ELIPs. Ultrasound was performed above the culture dish to break ELIPs and release the gas. After treatment, the expression level of the EGR2 gene, a marker of SC demyelination, was detected, and RNA was extracted for mRNA sequencing. Sequencing results were validated in SC and rat dorsal root ganglion (MRP) tissues through Western blot and qPCR. Results After Xe treatment, the level of EGR2 in SC cultured in vitro decreased, indicating that Xe treatment can promote the demyelination process of SC. RNA-sequencing results showed that after Xe treatment, 88 genes were up-regulated and 231 genes were down-regulated. Top 10 KEGG pathway enrichment analysis suggested an increase in MAPK pathway expression. Meanwhile, among the differentially expressed genes, we found that the expression of SPRY2 (Sprouty RTK Signaling Antagonist 2) decreased, a negative feedback regulator of the MAPK pathway. Using qPCR and Western blot on SC and rat MPG tissues in vitro, we validated the sequencing results and found that after Xe treatment, the expression level of SPRY2 decreased, and the level of pERK increased. Conclusions Xe may promote the demyelination of Schwann cells and the repair of cavernous nerves by inhibiting the SPRY2 level of SC, reducing its inhibition of the MAPK/ERK pathway. Disclosure No.

A Novel Multi-compartment Rotating Bioreactor for Improving ADSC--Spheroid Formation and its Application in Neurogenic Erectile Dysfunction.

The aim of this study was to construct a multicompartment synchronous rotating bioreactor (MCSRB) for batch-production of homogenized adipose-derived stem cell (ADSC) microspheres and treat neurogenic erectile dysfunction (ED). Firstly, an MCSRB was constructed using a centrifugal device and hinged trays. Secondly, influence factors (density, rotational speed) on the formation of ADSC-spheroids were explored. Finally, a neurogenic ED model was established to verify the effectiveness and safety of ADSC-spheroids for ED treatment. An MCSRB promoted ADSCs to gather microspheres, most of which were 90-130 μm in diameter. Supernatant from three-dimensional culture led to a significant increase in cytokine expression in ADSCs and migration rate in human umbilical vein endothelial cells (HUVECs) compared to control groups. The erectile function and pathological changes of the penis were improved in the ADSC-spheroids treatment group compared to the traditional ADSCs treatment group (p < 0.01). Efficient, batch, controlled and homogenized production of ADSC stem cell microspheres, and effective improvement of erectile dysfunction in neurogenic rats can be achieved using the MCSRB device.

Nrf2 enhances the therapeutic efficiency of adipose-derived stem cells in the treatment of neurogenic erectile dysfunction in a rat model

BackgroundErectile dysfunction (ED) caused by intraoperative nerve injury is a major complication of pelvic surgery. Adipose-derived stem cells (ADSCs) have presented therapeutic potential in a rat model of bilateral cavernous nerve injury (BCNI), while inadequate in vivo viability has largely limited their application. Nuclear factor-E2-related Factor (Nrf2) is a key transcription factor that regulates cellular anti-oxidative stress. In this work, we investigated the effect of Nrf2 expression regulation on the viability of ADSCs, and explore its repair potential in a BCNI rat model.ResultsThe survival time of tert-Butylhydroquinone (tBHQ)-ADSCs in BCNI model increased obviously. In addition, the tBHQ-ADSCs group presented better restoration of major pelvic ganglion (MPG) nerve contents and fibers, better improvement of erectile function, and less penile fibrosis than the other groups. Moreover, the expression of Nrf2 and superoxide dismutase 1 (SOD1) were higher than those of other groups.ConclusionNrf2 could enhance the anti-oxidative stress ability of ADSCs, so as to improve the therapeutic effect of ADSCs on BCNI rat model.

Ultrasound-targeted microbubble destruction mediates PDE5i/NO integration for cavernosum remodeling and penile rehabilitation.

Erectile dysfunction (ED) caused by cavernous nerve injury (CNI) is refractory to heal mainly ascribed to the adverse remodeling of the penis induced by ineffectual microvascular perfusion, fibrosis, and neurotrophins scarcity in cavernosum. Phosphodiesterase type V inhibitors (PDE5i) have been regarded as an alternative candidate drug for avoiding penile neuropathy. However, the therapeutic efficacy is severely limited due to poor accumulation under systemic medication and endogenous nitric oxide (NO) deficiency in cavernosum. Herein, an innovative liposomal microbubble (MB) loaded with both Sildenafil (one of PDE5i) and NO was designed. Ultrasound-targeted MB destruction (UTMD)-mediated efficient release and integration erectogenic agents into corpus cavernosum with high biosafety. On a bilateral CNI rat model, the multifunctional MB-cooperated UTMD improved microvascular perfusion in penis, simultaneously, alleviated hypoxia and oxidative stress, indicating successful activation of NO-cyclic guanosine monophosphate pathway. Also, evaluation of the endothelial/muscular composition, intracavernosal pressure, and neural integrity in the penis proved that coordinated intervention reversed the abnormal structural remodeling and promoted the recovery of functional erection. Our work demonstrates that MB loading Sildenafil and NO combined with UTMD hold great promise to "awaken" the efficacy of PDE5i in neurogenic ED, which provided a superior option for ensuring penile rehabilitation.

(093) The Light-Controlled Nitric Oxide Donor “NORD-1” may Improve Erectile Function Without Hypotension and Tissue Damage in Rats With Neurogenic Erectile Dysfunction

Abstract Introduction Radical prostatectomy (RP) often causes erectile dysfunction (ED) because of cavernous nerve damage during surgery. This damage results in a decrease of nitric oxide (NO) in the penile corpus cavernosum. Therefore, NO supplementation is considered to be effective for post RP ED. However, NO has systemic side effects such as headache and hypotension. Recently, a red-light-controlled NO donor “NORD-1”, developed by our group, enables spatially and temporally controlled NO release by light irradiation. Our previous study reported that NORD-1 combined with red-light irradiation is effective for neurogenic ED model rats with bilateral cavernous nerve injury (BCNI). However, the side effects of the treatment have not yet been discussed. Objective The study aimed to investigate the potency of side effects associated with the combination of NORD-1 and red-light irradiation, using a ED rat model with BCNI. Methods In this study, eight-week-old male Wistar-ST rats were used. The rats were divided into four groups: Sham+SiR650 (control compound), Sham+NORD-1, BCNI+SiR650, and BCNI+NORD-1 (n=6). The sham or BCNI surgery was performed under anesthesia in each group. BCNI surgery was performed by crushing the cavernous nerves bilaterally. Four weeks post-surgery, erectile function before and after the injection of each reagent into the penile corpus cavernosum was evaluated based on the changes in intracavernous pressure (ICP) under electrostimulation of the cavernous nerve. After the administration of each reagent, the changes in ICP with and without light irradiation were also evaluated. ICP during electrostimulation was totally measured three times; once without reagent and light, once with reagent and without light and once with reagent and light. The variations in systemic arterial pressure during electrostimulation were also analyzed. After the experiment, penile samples were collected to observe NORD-1 localization using a fluorescence microscope. In addition, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was performed to evaluate apoptosis in the corpus cavernousm. Results Before administering each reagent without light irradiation, the ICP/mean arterial pressure (MAP) ratios under stimulation in the BCNI+SiR650 and BCNI+NORD-1 groups were significantly lower than those in the Sham+SiR650 (P &amp;lt; 0.01) and Sham+NORD-1 (P &amp;lt; 0.05) groups. After administering each reagent without light irradiation, the ICP/MAP ratios in the BCNI+SiR650 and BCNI+NORD-1 groups remained significantly lower than those in the Sham+SiR650 and Sham+NORD-1 group (P &amp;lt; 0.05 and P &amp;lt; 0.01, respectively). After administering each reagent with light irradiation, the ICP/MAP ratios in the Sham+NORD-1 and BCNI+NORD-1 groups were significantly higher than those in the Sham+SiR650 (P &amp;lt; 0.01) and BCNI+NORD-1 (P &amp;lt; 0.01) groups, respectively. The changes of MAP during electrostimulation were not different under three different conditions in all four groups. Fluorescence of NORD-1 was detected only in corpus cavernousum which means it was not detected in urethra and dorsal vein of penis. Apoptosis was not detected in corpus cavernousum in all four groups. Conclusions The combination of NORD-1 and light irradiation is effective for neurogenic ED in rats with cavernous nerve injury. The treatment may not cause hypotension and tissue damage. Disclosure No

(229) Effect of Filtrated Bone Marrow-Derived Stem Cell Lysate on Erectile Function and Nerve Regeneration in Rats With Neurogenic Erectile Dysfunction

Abstract Introduction The transplantation of bone marrow-derived mesenchymal stem cell (BMSC) is believed to be a novel therapy for erectile dysfunction (ED) following radical prostatectomy (RP). However, the difficulty in preparation and higher costs challenge clinical application of this therapy. To overcome these challenges, we investigated the feasibility of filtrated BMSC lysate (BSCL) to alleviate ED in rats. We have reported BSCL improved erectile function in a rat model of bilateral cavernous nerve injury (BCNI). However, the mechanism remains unclear. Objective The study aimed to investigate the mechanism of BSCL for improving neurogenic ED focusing on nerve regeneration. Methods BSCL was obtained by freeze fracturing and filtrating BMSCs which were obtained from Wistar/ST rats. Three experiments were performed in this study. The first experiment included an eight-week-old male Wistar/ST rats, further divided into three groups: Sham + PBS, BCNI + PBS, and BCNI + BSCL (n=8). Rats in the Sham + PBS group underwent sham surgery, while those in the BCNI + PBS group underwent BCNI surgery. Immediately after the surgery, phosphate-buffered saline (PBS) was injected into the corpus cavernosum. Rats in the BCNI+BSCL group also underwent BCNI surgery. Immediately after the surgery, BSCL was injected into the corpus cavernosum. Four weeks after the surgery, erectile function was evaluated by measuring intracavernosal pressure (ICP)/mean arterial pressure (MAP) under stimulation of cavernous nerve. The second experiment was conducted on eight-week-old male Wistar/ST rats, further divided into three groups, same as in the first experiment. Three weeks after the surgery, fluorogold (FG) was injected into the corpus cavernosum from both sides. One week after the injection, bilateral major pelvic ganglions (MPGs) were dissected. FG reaching to MPG was observed using fluorescence microscope. The MPGs were also proved with anti PGP9.5, a neuron cell marker. In the third experiment, MPG were dissected from ten-week-old male Wistar/ST rats, placed on Matrigel and incubated with normal medium or medium including BSCL (50, 125, 250, and 500 ng/mL, respectively). After 48 hours, neurite outgrowth was measured using a Z-stack microscope. Results ICP/MAP in the BCNI+PBS group was significantly lower than that in the Sham+PBS group (P &amp;lt; 0.05), and ICP/MAP in the BCNI+BSCL group was significantly higher than that in the BCNI+PBS group (P &amp;lt; 0.05). The ratios of FG positive cells to PGP9.5 positive cells in the BCNI+PBS group were significantly lower than those in the Sham+PBS group. Those in the BCNI+BSCL group were higher than those in the BCNI+PBS group. MPG treated with the medium including BSCL had longer neurite outgrowth than MPG treated with normal medium. This neurite outgrowth effect of BSCL increased in a dose-dependent manner. Conclusions In conclusion, BSCL treatment improved erectile function in rats with neurogenic ED. In addition, BSCL has the potential to improve neurotransmission of cavernous nerves and BSCL exhibited neurite outgrowth effects on rat MPG. The study demonstrates that BSCL can regenerate a crushed cavernous nerve, improving erectile function. Further study should be conducted to identify active factors in BSCL contributing to nerve regeneration. Disclosure No

(094) Synergy Between Orally Administered Phosphodiesterase-type-5-inhibitors and Topically Delivered Nitric-oxide in Eliciting Erectile Responses

Abstract Introduction Phosphodiesterase-type-5-inhibitors (PDE5i) represent first-line FDA-approved oral treatments for erectile dysfunction (ED). However, because their mode of action is dependent on the production of nitric oxide (NO) from nerve endings, patients that suffer neurogenic ED (for example, following the procedures for radical prostatectomy (RP) or resulting from diabetes) are commonly refractory to PDE5i treatment. We recently demonstrated proof-of-principle for increased efficacy of topically applied NO-releasing microparticles (NO-MP) in eliciting an erectile response when used in combination with the orally administered PDE5i, sildenafil. However, there are several other FDA approved PDE5i used to treat ED, which, although acting through the common mechanism of PDE5 inhibition, exhibit several characteristics that potentially could impact clinical and commercial translation of combination therapy. Objective The goal of the present studies was to determine if other commercially available PDE5i besides sildenafil (avanafil, tadalafil and vardenafil) also act cooperatively when used in combination with NO-MP to elicit an erectile response. Methods Sprague-Dawley rats underwent bi-lateral transection of the cavernous nerve (CN), as a model of RP, one-week prior to determining erectile response to treatment. We investigated the effect of oral administration of 0.05mg sildenafil/kg, 0.005mg tadalafil/kg, 0.01mg vardenafil/kg and 0.1mg avanafil/kg (approximately equivalent to clinical doses of 2.5mg, 0.25mg, 0.5mg and 5mg, respectively, administered to an 80 kg patient) 30 minutes prior to being anesthetized for the determination of erectile response by intracorporal pressure (ICP) and systemic blood pressure (BP). Baseline ICP and BP were recorded for 30 minutes, at which time 100 mg of NO-MP (or control, blank-MP) were applied to the rat penile shaft. The ICP and systemic BP were used to determine the time to the first erectile response, duration of the erectile responses, the maximal and average ICP/BP responses and the number of spontaneous erections per hour. Results Although oral treatment by PDE5i alone resulted in no observable erectile response, topical application of NO-MP resulted in spontaneous erectile responses. When NO-MP were applied in combination with oral PDE5i treatment, all PDE5i resulted in faster times to the first erectile response (on average 11 minutes compared to 22 minutes when NO-MP were used alone; P-value=0.041) and greater numbers of spontaneous erections (on average 4.6 erections per hour compared to 2.0 when NO-MP were used alone, P-value=0.043). Although there were no significant differences between any of the PDE5i when used in combination with NO-MP with respect to the average erectile responses per hour, duration of erectile response, and maximal ICP/BP, there was a trend in the vardenafil and avanafil treated animals for a more rapid onset of the erectile responses than observed with sildenafil and tadalafil. Conclusions Although knowledge of the molecular pathways that lead to an erection provide a rationale for a synergistic effect between PDE5i and NO in eliciting an erectile response, our results provide the first evidence of such synergy. Our results suggest a novel treatment strategy for patients with ED refractory to PDE5i, namely any of the FDA-approved oral PDE5 inhibitors used in combination with formulations that topically deliver NO to corporal tissue. Disclosure Yes, this is sponsored by industry/sponsor: Zylo Therapeutics, Inc Clarification Industry funding only - investigator initiated and executed study Any of the authors act as a consultant, employee or shareholder of an industry for: Zylo Therapeutics, Inc

LIPUS-SCs-Exo promotes peripheral nerve regeneration in cavernous nerve crush injury-induced ED rats via PI3K/Akt/FoxO signaling pathway.

Clinical treatment of erectile dysfunction (ED) caused by cavernous nerve (CN) injury during pelvic surgery is difficult. Low-intensity pulsed ultrasound (LIPUS) can be a potential strategy for neurogenic ED (NED). However, whether Schwann cells (SCs) can respond to LIPUS stimulation signals is unclear. This study aims to elucidate the signal transmission between SCs paracrine exosome (Exo) and neurons stimulated by LIPUS, as well as to analyze the role and potential mechanisms of exosomes in CN repair after injury. The major pelvic ganglion (MPG) neurons and MPG/CN explants were stimulated with LIPUS of different energy intensities to explore the appropriate LIPUS energy intensity. The exosomes were isolated and purified from LIPUS-stimulated SCs (LIPUS-SCs-Exo) and non-stimulated SCs (SCs-Exo). The effects of LIPUS-SCs-Exo on neurite outgrowth, erectile function, and cavernous penis histology were identified in bilateral cavernous nerve crush injury (BCNI)-induced ED rats. LIPUS-SCs-Exo group can enhance the axon elongation of MPG/CN and MPG neurons compared to SCs-Exo group in vitro. Then, the LIPUS-SCs-Exo group showed a stronger ability to promote the injured CN regeneration and SCs proliferation compared to the SCs-Exo group in vivo. Furthermore, the LIPUS-SCs-Exo group increased the Max intracavernous pressure (ICP)/mean arterial pressure (MAP), lumen to parenchyma and smooth muscle to collagen ratios compared to the SCs-Exo group in vivo. Additionally, high-throughput sequencing combined with bioinformatics analysis revealed the differential expression of 1689 miRNAs between the SCs-Exo group and the LIPUS-SCs-Exo group. After LIPUS-SCs-Exo treatment, the phosphorylated levels of Phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt) and forkhead box O (FoxO) in MPG neurons increased significantly compared to negative control (NC) and SCs-Exo groups. Our study revealed that LIPUS stimulation could regulate the gene of MPG neurons by changing miRNAs derived from SCs-Exo, then activating the PI3K-Akt-FoxO signal pathway to enhance nerve regeneration and restore erectile function. This study had important theoretical and practical significance for improving the NED treatment.

Role of mechanotransduction mediated by YAP/TAZ in the treatment of neurogenic erectile dysfunction with low-intensity pulsed ultrasound.

Erectile dysfunction (ED) and weakness of the penis are processes related to hemodynamic alteration. Low-intensity pulsed ultrasound (LIPUS), as a new mechanical modality for the treatment of ED, deserves to be explored in depth for the biomechanical mechanisms it exerts. The aim of this study was to explore the role of YAP/TAZ-mediated mechanotransduction in mechanical therapy for the treatment of neurogenic erectile dysfunction (NED). Forty-two male SD rats (12 w old) were randomly divided into sham-operated (n=14), bilateral cavernous nerve injury (BCNI, n=14), and LIPUS-treated (n=14) groups. Intracavernosal pressure/mean arterial pressure (ICP/MAP) was measured 14 and 28 days after treatment. Penile tissue specimens were collected for pathological examination, and the changes in YAP, TAZ, connective tissue growth factor (CTGF), CYR61, LATS1, and p38 mitogen-activated protein kinase expression levels were assessed by Western blot, real-time quantitative polymerase chain reaction (RT-qPCR) and immunological staining. Compared with BCNI, LIPUS significantly improved ICP/MAP levels and enhanced histopathological changes. The penile expression levels of YAP, TAZ, CTGF, and CYR61 were significantly downregulated in the BCNI group (p<0.01), and LIPUS upregulated the expression levels of these proteins (p<0.05). The expression levels of p-LATS1 and LATS1 were not significantly different among the groups (p>0.05). Interestingly, the expression level of p-p38/p38 significantly increased in BCNI rats (p<0.05), which was reversed by LIPUS treatment (p<0.05). However, the p38 inhibitor SB203580 did not change the expression of YAP/TAZ in rat primary smooth muscle cells or mouse MOVAS cells (p>0.05). LIPUS can effectively improve penile erectile function in NED rats. The underlying mechanism may be related to the regulation of YAP/TAZ-mediated mechanotransduction. However, the upstream regulatory signal may differ from the classical Hippo pathway.

Low-intensity pulsed ultrasound ameliorates erectile dysfunction induced by bilateral cavernous nerve injury through enhancing Schwann cell-mediated cavernous nerve regeneration.

Cavernous nerve injury-induced erectile dysfunction caused by pelvic surgery or trauma is refractory to conventional medications and required an alternative treatment. Low-intensity pulsed ultrasound is a noninvasive mechanical therapy that promotes nerve regeneration. To investigate the therapeutic effect and potential mechanism of low-intensity pulsed ultrasound in the treatment of neurogenic erectile dysfunction. Thirty rats were randomly divided into the sham-operated group, bilateral cavernous nerve injury group, and bilateral cavernous nerve injury + low-intensity pulsed ultrasound group. The erectile function was assessed 3weeks after daily low-intensity pulsed ultrasound treatment. The penile tissues and cavernous nerve tissues were harvested and subjected to histologic analysis. Primary Schwann cells and explants were extracted from adult rats. The effects of low-intensity pulsed ultrasound on proliferation, migration, and nerve growth factor expression of Schwann cells and axonal elongation were examined in vitro. RNA sequencing and western blot assay were applied to predict and verify the molecular mechanism of low-intensity pulsed ultrasound-induced Schwann cell activation. Our study showed that low-intensity pulsed ultrasound promoted Schwann cells proliferation, migration, and neurotrophic factor nerve growth factor expression. Meanwhile, low-intensity pulsed ultrasound exhibits a stronger ability to enhance Schwann cells-mediated neurite outgrowth of major pelvic ganglion neurons and major pelvic ganglion/cavernous nerve explants in vitro. In vivo experiments demonstrated that the erectile function of the rats in the bilateral cavernous nerve injury + low-intensity pulsed ultrasound group was significantly higher than those in the bilateral cavernous nerve injury groups. Moreover, the expression levels of smooth muscle and cavernous endothelium also increased significantly in the bilateral cavernous nerve injury + low-intensity pulsed ultrasound group. In addition, we observed the higher density and number of cavernous nerve regenerating axons in the bilateral cavernous nerve injury + low-intensity pulsed ultrasound group, indicating that low-intensity pulsed ultrasound promotes axonal regeneration following cavernous nerve injury in vivo. RNA sequencing analysis and bioinformatic analysis suggested that low-intensity pulsed ultrasound might trigger the activation of the PI3K/Akt pathway. Western blot assay confirmed that low-intensity pulsed ultrasound activated Schwann cells through TrkB/Akt/CREB signaling. Low-intensity pulsed ultrasound promoted nerve regeneration and ameliorated erectile function by enhancing Schwann cells proliferation, migration, and neurotrophic factor nerve growth factor expression. The TrkB/Akt/CREB axis is the possible mechanism of low-intensity pulsed ultrasound-mediated Schwann cell activation. Low-intensity pulsed ultrasound-based therapy could be a novel potential treatment strategy for cavernous nerve injury-induced neurogenic erectile dysfunction.

Riluzole, a neuroprotective agent, preserves erectile function following bilateral cavernous nerve injury in male rats.

Neurogenic erectile dysfunction is a highly prevalent complication in men undergoing radical prostatectomy. The underlying mechanisms remain incompletely defined and the effective therapy has been limited. This study aimed to evaluate the protective effect of riluzole and the role of PKC β and excitatory amino acid transporters (EAATs) mediating this effect in a rat model of bilateral cavernous injury (BCNI). A total of 48 male Sprague-Dawley rats were divided into sham, BCNI (at 7, 15 days post-injury) and BCNI treated with riluzole (8 mg/kg/day) groups. Erectile function was measured as maximum intracavernosal pressure (mICP)/mean arterial pressure (MAP) and total ICP/MAP. Changes in protein expressions of phospho (p)-PKC β IIser660 and EAATs were analysed in penis and major pelvic ganglion with western blotting. BCNI decreased erectile function at 7 and 15 days post-injury (mICP/MAP at 4 V: 0.45 ± 0.06 vs 0.84 ± 0.07; 0.34 ± 0.04 vs 0.77 ± 0.04 respectively; p < 0.001) whereas riluzole treatment (for 15 days) preserved erectile function (mICP/MAP at 4 V: 0.62 ± 0.03 vs 0.34 ± 0.04; p < 0.01). The decline in the expression of p-PKC β IIser660 was observed in penis at 7 and 15 days post-injury (p = 0.0003, p = 0.0033), which was prevented by riluzole treatment for 15 days (p = 0.0464). While expressions of EAAT-1 and EAAT-2 decreased in major pelvic ganglion following BCNI (p = 0.0428, p = 0.002), riluzole treatment for 15 days prevented the decrease only in EAAT-2 expression (p = 0.0456). Riluzole improved erectile function via possibly interacting with PKC β II and glutamatergic pathways, as a potential therapeutic candidate for erectile dysfunction.

Overexpression of PRDX2 in Adipose-Derived Mesenchymal Stem Cells Enhances the Therapeutic Effect in a Neurogenic Erectile Dysfunction Rat Model by Inhibiting Ferroptosis.

Neurogenic erectile dysfunction (NED) is a common and serious complication after pelvic surgery. The clinical translation of adipose-derived mesenchymal stem cell (ADSC) therapies in NED remains a major challenge due to their low survival rate and limited therapeutic effect. Peroxiredoxin 2 (PRDX2) is a member of the peroxidase family that exerts its therapeutic effects by inhibiting oxidative stress (OS) and ferroptosis, and PRDX2 is expected to enhance the therapeutic effect of ADSCs in treating NED. The purpose of this study was to investigate whether PRDX2 could improve the survival of ADSCs and determine whether overexpression of PRDX2 in ADSCs (PRDX2-ADSCs) could enhance the therapeutic effect of NED. This study investigated the potential role of PRDX2-ADSCs through a NED model induced by bilateral cavernous nerve injury (BCNI) and three in vitro models established by H2O2-stimulated ADSCs, H2O2-stimulated corpus cavernosum smooth muscle cells (CCSMCs), and RSL3-stimulated CCSMCs. We found that PRDX2 could significantly improve the viability of ADSCs by suppressing apoptosis and OS in H2O2-stimulated ADSCs. We also found that BCNI triggered ferroptosis of the corpus cavernosum, which was manifested by increased reactive oxygen species (ROS), total iron content, and MDA as well as decreased SOD and GSH. Our results further demonstrated changes in the expression of key proteins (GPX4 and ACSL4) in the ferroptosis pathway, whereas PRDX2-ADSCs ameliorated BCNI-induced erectile dysfunction and ferroptosis of the corpus cavernosum in NED rats. Consistently, PRDX2-ADSCs attenuated OS in H2O2-stimulated CCSMCs and inhibited ferroptosis in RSL3-stimulated CCSMCs, as evidenced by the decrease in ROS, total iron content, and MDA and the increase in SOD and GSH together with changes in ferroptosis-related protein (GPX4 and ACSL4) expression. In conclusion, overexpression of PRDX2 in ADSCs enhanced the therapeutic effect in a rat model of neurogenic erectile dysfunction by inhibiting ferroptosis via regulation of the GPX4/ACSL4 axis.

Melatonin-Primed Mesenchymal Stem Cells-Derived Small Extracellular Vesicles Alleviated Neurogenic Erectile Dysfunction by Reversing Phenotypic Modulation.

Erectile dysfunction (ED) is an adverse side effect of pelvic surgery with no effective treatment. In this study, it is explored whether melatonin could improve the therapeutic effects of small extracellular vesicles (sEVs), derived from mesenchymal stem cells (MSCs), on cavernous nerve injury (CNI) ED, and the underlying mechanisms are investigated. The sEVs from melatonin-pretreated MSCs (MT-EVs) and MSCs (NC-EVs) are isolated and applied to CNI ED. Transplantation of MT-EVs remarkably increases erectile function and reduces phenotypic modulation in CNI ED rats. The therapeutic effects of MT-EVs are superior to those of NC-EVs. Sequencing implies that miR-10a-3p is enriched in MT-EVs, and directly targets the protein kinase inhibitor α (PKIA). After the suppression of miR-10a-3p, the therapeutic actions of MT-EVs are abolished, but are rescued by PKIA. Similarly, RhoA/ROCK is inhibited by MT-EVs, but this action is reversed by suppressing miR-10a-3p, accompanied by corresponding changes in PKIA. In conclusion, transplantation of MT-EVs could significantly alleviate CNI ED. MT-EVs may relieve the phenotypic modulation of the corpora cavernosum smooth muscle cells via the miR-10a-3p/PKIA/RhoA/ROCK signaling axis. These nanovesicles should be potential therapeutic vectors or bioactive materials for CNI ED.

In vitro high glucose increases apoptosis, decreases nerve outgrowth, and promotes survival of sympathetic pelvic neurons.

Diabetes mellitus (DM) is a common cause of erectile dysfunction (ED), yet the molecular basis of DM neurogenic ED remains unknown. In this study we examined the impact of high glucose on survival and growth of primary cultured pelvic neurons in a rat model and assessed whether coculturing with healthy Schwann cells (SCs) can rescue pelvic neuron growth in patients with DM. Major pelvic ganglia (MPGs) from adult male Sprague Dawley rats (n = 8) were dissociated and plated on coverslips. Neurons were exposed to high glucose (45mM) for 24 or 48hours and compared to time-matched controls (25mM). Neurons were stained for neuron-specific beta-tubulin, neuronal nitric oxide synthase, vesicular acetylcholine transferase, tyrosine hydroxylase, and TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling) assay. Schwann cells were dissociated from MPGs of healthy male Sprague Dawley rats (n = 4) and grown to confluence. Additional Sprague Dawley rats were made diabetic with streptozotocin (50mg/kg, n = 4), and 5weeks later MPGs were collected from these rats, dissociated, and cocultured on healthy SCs. Neurons and SCs were stained with beta-tubulin and S100. Length, branching, and survival of nitrergic, parasympathetic, and sympathetic neurons was assessed in neurons exposed to normal or high glucose concentrations, and neuron length was measured in neuron-SC coculture. The total number of neurons and the length and number of branches were significantly decreased after 24 and 48hours of high glucose (P < .05). The percentage of nitrergic neurons decreased 10% after 24 hours and 50% after 48 hours of high glucose (P < .05). After 24 hours of high glucose, cholinergic-positive neurons were unchanged; however, these neurons decreased 30% after 48 hours (P < .05). The proportion of sympathetic neurons increased 25% after 48 hours of high glucose (P < .05). At both timepoints, there was a 2-fold increase in the total apoptotic neurons with high glucose (P < .05). Neurite outgrowth recovered to control lengths after coculture of diabetic neurons with healthy SCs (P < .05). Glucose can be used as a tool to investigate the direct effects of DM on neuritogenesis. Our data suggest that an effective treatment for DM ED protects and repairs the penile neuronal supply. Exposing MPG neurons to high glucose offers a quick and, inexpensive proxy for DM-related conditions. A limitation of our study is that our model reflects type 1 DM, whereas clinically, most diabetic ED patients have type 2 DM. Culturing pelvic neurons in high glucose can be used as a tool to elucidate how to protect proerectile neurons from cell death and may lead to new therapeutic strategies for diabetic men suffering from ED.

Efficacy of a Red-Light Controllable Nitric Oxide Releaser for Neurogenic Erectile Dysfunction: A Study Using a Rat Model of Cavernous Nerve Injury.

Neurogenic erectile dysfunction (ED) is a common side effect of radical prostatectomy (RP) because of cavernous nerve damage. In these patients, the production of nitric oxide (NO), which is important for erection, is decreased in the corpus cavernosum. Therefore, NO donors are useful for post-RP ED. However, short half-life and systemic side effects are problems of NO application in ED therapy. To avert these problems, we developed a red-light controllable NO releaser, NORD-1. This study aimed to investigate the effect of NORD-1 and red-light irradiation on neurogenic ED using a rat model of bilateral cavernous nerve injury (BCNI). BCNI and sham operations were conducted on 8-week-old rats. After 4 weeks, erectile function was evaluated using changes in intracavernous pressure (ICP) during electrostimulation of the cavernous nerve. ICP was measured under three conditions; without NORD-1 and red-light irradiation, with NORD-1 and without red-light irradiation, and with NORD-1 and red-light irradiation. SiR650 which absorbs red-light but does not release NO was used for the negative control. After the experiment, localization of NORD-1 was observed using a microscope. Erectile function in a BCNI rat model was significantly decreased compared to sham-operated rats (p<0.05). After injecting NORD-1 into the penis, erectile function did not change without red-light irradiation. However, the combination of NORD-1 and red-light irradiation significantly improved erectile function (p<0.05) without affecting systemic arterial pressure. In contrast, when SiR650 was used, erectile function did not change in all three conditions. NORD-1 was detected only in the corpus cavernosum and not in the urethra and dorsal vein. NORD-1 combined with red-light irradiation is effective for ED induced by cavernous nerve injury. This treatment may have low risks of hypotension and urinary incontinence, and it can replace the current treatment for post-RP ED.

Improved erectile function after focal muscle vibrations therapy in a patient with neurogenic erectile dysfunction: a case report

To report a case of improved Erectile Dysfunction (ED) after surgery for colorectal cancer using focal Muscle Vibrations (MV). A 59-year-old male with neurogenic ED following surgery for colorectal cancer 18 months ago with an International Index of Erectile Function (IIEF) score <18, sexually active, fully continent before surgery; and withdrawal from previous treatment for at least three months such as erectogenic aids, antispastics, and psychoactive drugs, was encouraged to use a male vibrator called Tenuto®, three times a week for five consecutive weeks, each session lasting 30 min. The stimulation was delivered to the perineum and suprapubic areas at a frequency of 130 Hz. Changes in the validated erectile function domain of the IIEF were used to estimate the effectiveness of MV in improving ED. It has been suggested that an increase in of 6 points in this domain indicates a clinical improvement. At the end of five weeks, the patient showed improvement over baseline of 8 points for the erectile function domain scores from 7 to 15 with using the device. Although patients experiencing neurogenic ED are a multifaceted group, in this case we believe that MV improves ED through improved spasticity. Previous research shows that muscle MV may improve muscle synergies by counterbalancing the spasticity. Future studies using large sample sizes with longer follow-up and group control could provide more evidence in order to confirm this hypothesis. i have no conflict of interest to declare

Improved erectile function after focal muscle vibrations therapy in a patient with neurogenic erectile dysfunction: a case report

Improved erectile function after focal muscle vibrations therapy in a patient with neurogenic erectile dysfunction: a case report

Co-overexpression of VEGF and Smad7 improved the therapeutic effects of adipose-derived stem cells on neurogenic erectile dysfunction in the rat model.

Cavernous nerve injury is the main cause of erectile dysfunction (ED) after radical prostatectomy (RP). In our previous study, injection of adipose-derived stem cells (ADSCs) into the cavernosum can repair damaged cavernosum nerves and ED can be restored to a certain extent. In order to improve these therapeutic effects, we evaluated the efficacy of ADSCs co-modified with VEGF and Smad7 in a rat model. SD rats were randomly divided into six groups: a sham surgery group, and the five bilateral cavernous nerve injury (BCNI) groups were injected with ADSC or ADSCs genetically modified by VEGF (ADSC-V), Smad7 (ADSC-S), or VEGF&Smad7 (ADSC-V&S) or phosphate-buffered saline (PBS). The results indicated that the erectile function of the ADSC-V, ADSC-S, and ADSC-V&S groups was significantly recovered, and the erectile function of the ADSC-V&S group was more distinctly recovered as compared to the other groups. The same results are shown in the expression of neuronal nitric oxide synthase and the smooth muscle/collagen ratio of penile tissue comparing the ADSC-V&S group to the ADSC-V and ADSC-S group. These experimental data suggest that ADSCs co-overexpressed with VEGF and Smad7 can significantly improve erectile function after BCNI. This study provides new therapeutic thoughts for ED following RP.