Bilateral Cavernous Nerve Injury Research Articles

Intracavernous Pressure Recording in a Cavernous Nerve Injury Rat Model.

The bilateral cavernous nerve (CN) injury rat model has been extensively used to simulate clinical cavernous nerve injury associated with erectile dysfunction (ED) for evaluating the effect of clinical therapeutic methods. However, the methods of CN injury model construction are flawed and varied in the ED research field. It is CN crush injury that is the most commonly used method in recent years. This study aims to provide a detailed description of the procedure of bilateral CN injury rat model construction and measurement of intracavernous pressure (ICP) recording, providing a reliable and reproducible CN injury rat model. This work successfully developed the CN injury method of hemostat crush injury using a syringe needle as hard support and a hemostat with a rubber sleeve. Also, this method concludes that a voltage of 1.0 V, frequency of 20 Hz, and pulse-width of 5 ms are the optimized stimulation parameters for ICP recording in a bilateral CN injury rat model.

PD62-07 TUMOR NECROSIS FACTOR ALPHA (TNF- Α) INHIBITOR IMPROVED ERECTILE FUNCTION IN A BILATERAL CAVERNOUS NERVE INJURY RAT MODEL

You have accessJournal of UrologySexual Function/Dysfunction: Basic Research & Pathophysiology (PD62)1 Sep 2021PD62-07 TUMOR NECROSIS FACTOR ALPHA (TNF- Α) INHIBITOR IMPROVED ERECTILE FUNCTION IN A BILATERAL CAVERNOUS NERVE INJURY RAT MODEL James Liu, Hotaka Matsui, Kara Lombardo, Serkan Karakus, Arthur Burnett, and Trinity Bivalacqua James LiuJames Liu More articles by this author , Hotaka MatsuiHotaka Matsui More articles by this author , Kara LombardoKara Lombardo More articles by this author , Serkan KarakusSerkan Karakus More articles by this author , Arthur BurnettArthur Burnett More articles by this author , and Trinity BivalacquaTrinity Bivalacqua More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002099.07AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Erectile dysfunction (ED) after injury to the peripheral cavernous nerves (CN) is partially the result of an increase in neuroinflammation in the pelvic ganglia. Inflammatory cytokines have been implicated in axonal degeneration and neuropathy. This study examines the role of the inflammatory cytokine, tumor necrosis factor –alpha (TNF- α), after bilateral CN injury (BCNI) in vivo. We also investigate the effect of commercially available TNF- α inhibitors (Infliximab) on erectile function restoration in the BCNI rodent model. METHODS: Seventy male Sprague Dawley rats were randomized to undergo either BCNI or sham surgery. Sham rats major pelvic ganglion (MPG) were harvested after 48 hours, whereas the BCNI groups had MPG harvested at 6 hours, 12 hours, 24 hours, 48 hours, 7 days, or 14 days after BCNI. MPGs were analyzed by qPCR to evaluate gene expression of TNF-α, TNF- αR1, TNF- αR2 and markers of macrophages (CD11b and CD68). Western blot of MPGs were also performed to confirm these finding. Additionally, 25 Sprague Dawley rats were randomized to three groups: sham surgery, BCNI, or BCNI treated with TNF- α inhibitor (Infliximab) (5mg/kg weekly) for two weeks and then underwent cavernous nerve stimulation and erectile parameters were measured (peak ICP, ICP/MAP, and total ICP). RESULTS: BCNI significantly increased (p<0.05) gene and protein expression of TNF- α and its receptors (R1, R2) in rat MPGS, with peak levels were observed at 2 and 7 days, respectively. Corresponding macrophage markers (CD11b and CD68) were significantly increased (p<0.05) following BCNI with maximal expression at 48 hours. Erectile response studies demonstrated that rodents treated with TNF- α inhibitors had significantly improved (p<0.05) erectile function (ICP/MAP) at all voltages compared to BCNI alone (2V: 0.203 vs 0.057, 4V: 0.279 vs 0.081, 6V: 0.317 vs 0.166. Control 2V: 0.347, 4V: 0.414, 6V: 0.486). Peak ICP and total ICP were also significantly (p<0.05) improved compared to BCNI rats. CONCLUSIONS: TNF- α is a key inflammatory cytokine which contributes to neuroinflammation following BCNI. Treatment with commercially available TNF- α inhibitor results in mitigation of erectile dysfunction in a BCNI rat model when administered at the time of nerve injury. Source of Funding: None © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e1074-e1075 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information James Liu More articles by this author Hotaka Matsui More articles by this author Kara Lombardo More articles by this author Serkan Karakus More articles by this author Arthur Burnett More articles by this author Trinity Bivalacqua More articles by this author Expand All Advertisement Loading ...

Increased Level of Tumor Necrosis Factor-Alpha (TNF-α) Leads to Downregulation of Nitrergic Neurons Following Bilateral Cavernous Nerve Injury and Modulates Penile Smooth Tone

Erectile dysfunction (ED) after injury to peripheral cavernous nerve (CN) is partly a result of inflammation in pelvic ganglia, suggesting that ED may be prevented by inhibiting neuroinflammation. The aim of this study is to examine temporal changes of TNF-α, after bilateral CN injury (BCNI), to evaluate effect of exogenous TNF-α on neurite outgrowth from major pelvic ganglion (MPG), and to investigate effect of TNF-α signal inhibition to evaluate effects of TNF-α on penile tone with TNF-α receptor knockout mice (TNFRKO). Seventy Sprague-Dawley rats were randomized to undergo BCNI or sham surgery. Sham rats' MPGs were harvested after 48 hours, whereas BCNI groups' MPGs were at 6, 12, 24, 48 hours, 7, or 14 days after surgery. qPCR was used to evaluate gene expression of markers for neuroinflammation in MPGs. Western blot was performed to evaluate TNF-α protein amount in MPGs. MPGs were harvested from healthy rats and cultured in Matrigel with TNF-α. Neurite outgrowth from MPGs was measured after 3 days, and TH and nNOS immunofluorescence was assessed. Wild type (WT) and TNFRKO mice were used to examine effect of TNF-α inhibition on smooth muscle function after BCNI. MPGs were harvested 48 hours after sham or BCNI surgery to evaluate gene expression of nNOS and TH. Gene expression of TNF-α signaling pathway, Schwann cell and macrophage markers, protein expression of TNF-α in MPGs, and penile smooth muscle function to electrical field stimulation (EFS) were evaluated. BCNI increased gene and protein expression of TNF-α in MPGs. Exogenous TNF-α inhibited MPG neurite outgrowth. MPGs cultured with TNF-α had decreased gene expression of nNOS (P < .05). MPGs cultured with TNF-α had shorter nNOS+ neurites than TH+ neurites (P < .01). Gene expression of nNOS was enhanced in TNFRKO mice compared to WT mice (P < .01). WT mice showed enhanced smooth muscle contraction of penises of WT mice was enhanced to EFS, compared to TNFKO (P < .01). Penile smooth-muscle relaxation to EFS was greater in TNFKO mice compared to WT (P < .01). TNF-α inhibition may prevent ED after prostatectomy. TNF-α inhibition might prevent loss of nitrergic nerve apoptosis after BCNI and preserve corporal smooth muscle function but further investigation is required to evaluate protein expression of nNOS in MPGs of TNFKO mice. TNF-α inhibited neurite outgrowth from MPGs by downregulating gene expression of nNOS and TNFRKO mice showed enhanced gene expression of nNOS and enhanced penile smooth-muscle relaxation. Matsui H, Sopko NA, Campbell JD, etal. Increased Level of Tumor Necrosis Factor-Alpha (TNF-α) Leads to Downregulation of Nitrergic Neurons Following Bilateral Cavernous Nerve Injury and Modulates Penile Smooth Tone. J Sex Med 2021;18:1181-1190.

A novel flavonoid derivative of icariside II improves erectile dysfunction in a rat model of cavernous nerve injury.

Icariside II (ICA II), an active flavonoid monomer, has been proven to restore post-prostatectomy erectile dysfunction in rats; however, the high cost of extraction from natural plants limits the application of ICA II. To investigate the therapeutic effect and possible mechanism of action of YS-10, a new flavonoid compound, which was designed and synthesized based on the structure of ICA II in a rat model in of cavernous nerve injury. Eight of 32 adult male Sprague-Dawley rats were selected as the normal control (NC) group and received vehicle treatment. The remaining rats were subjected to bilateral cavernous nerve injury (BCNI) and randomized into three groups: BCNI group, BCNI + ICA II group (2.5mg/kg/day), and BCNI + YS-10 group (2.5mg/kg/day). The total procedure lasted for 21 days, followed by a washout period of 3 days. All animals were evaluated for erectile function, and tissues were harvested for histopathological analyses. It was observed that in YS-10 group, the ratio of intracavernous pressure (ICP) to mean arterial pressure (MAP) and the area under the ICP/MAP curve were effectively enhanced. The maximum ICP/MAP increased by 30% in the YS-10 group (0.86 ± 0.085) compared with the BCNI group (0.66 ± 0.058), which is close to 82% of the NC group (1.05 ± 0.033). Histopathological changes demonstrated significant reduction of smooth muscle atrophy, collagen deposition, and endothelial and neural dysfunction after YS-10 treatment, which have no statistical differences compared with ICA II group. Additionally, high-protein expression levels of β-Catenin and cyclin D1 were observed in the treatment groups. YS-10, a novel synthesized flavonoid compound, could effectively improve erectile dysfunction in rats after BCNI by alleviating pathological impairments; this effect may associate with the upregulation of β-Catenin and cyclin D1 in Wnt signaling pathway.

CXCL5 Cytokine Is a Major Factor in Platelet-Rich Plasma's Preservation of Erectile Function in Rats After Bilateral Cavernous Nerve Injury

BackgroundThe neuro-protective and tissue-protective properties of platelet-rich plasma (PRP) have been demonstrated through treating bilateral cavernous nerve (CN) injury in rats, although the underlying mechanisms have not been fully clarified. AimTo determine factors released from PRP and explore their role in mediating preservation of erectile function (EF) in a rat model of CN injury. MethodsMale Sprague-Dawley rats (aged 10 weeks) were used in this study. 6 rats were used to obtain blood for PRP and whole plasma preparation. We probed samples using a cytokine antibody array and performed enzyme-linked immunosorbent assay (ELISA). We determined the expression patterns of C-X-C motif chemokine ligand 5 (CXCL5) and receptors in the major pelvic ganglion (MPG) and corpus cavernosum via immunostaining. 32 rats were divided into 4 groups based on the type of injection received: (i) sham, (ii) vehicle, (iii) 400 μL of PRP, and (iv) 30 ng/kg of CXCL5. Groups 2–4 were subjected to bilateral CN crush (BCNC) injury. 4 weeks later, EF was assessed by CN electrostimulation, and CNs and penile tissue were collected for histological analysis. OutcomeCytokine antibody array, ELISA, erectile response, and immunofluorescence staining readings. ResultsThe PRP contained high levels of CXCL5. MPG neurons expressed CXCL5 and CXCR2. PRP intracavernous injection stabilized CXCR2 and increased CXCL5 expression in the MPG after BCNC, thus enhancing neuroprotection. CXCL5 injection improved BCNC-induced erectile dysfunction by preventing smooth muscle atrophy. Clinical ImplicationsThe therapeutic efficacy of PRP in CN injury–induced erectile dysfunction may arise from the synergy among multiple biomolecules. Our study serves as a basis for future studies on PRP formulation to provide safe and effective medications for the maintenance of EF after radical prostatectomy in patients with prostate cancer. Strengths & LimitationsA strength of our study is that our model was able to isolate the role of cytokines, specifically CXCL5, as part of the mechanism responsible for PRP's protective properties. However, the rat cytokine array provided limited experimental targets. The rats used were not at the age corresponding to prostate cancer patients in clinical settings. Our study did not explore CXCL5 blocking in the PRP group. Finally, the main protein quantification results by western blotting were hampered because of small tissue samples. ConclusionsThis study provides evidence for the role of CXCL5 and CXCR2 as mediators of PRP effects in the preservation of EF after CN injury.Wu YN, Liao CH, Chen KC, et al. CXCL5 Cytokine Is a Major Factor in Platelet-Rich Plasma's Preservation of Erectile Function in Rats After Bilateral Cavernous Nerve Injury. J Sex Med 2021;18:698–710.

Enhanced effects of salidroside on erectile function and corpora cavernosa autophagy in a cavernous nerve injury rat model.

We explored the efficacy and mechanisms of salidroside treatment for erectile dysfunction induced by bilateral cavernous nerve injury (BCNI). Forty male rats were divided into four groups as follows: sham (cavernous nerves exposed only) (S); BCNI (M); BCNI+rapamycin (M+rapamycin); and BCNI+salidroside (M+salidroside). Erectile function in the rats was measured by intracavernosal pressure. Penile tissue was harvested for transmission electron microscopy, immunohistochemistry, immunofluorescence, Masson's trichrome staining, haematoxylin-eosin staining, TdT-mediated dUTP Nick End Labeling and western blotting. The M group exhibited a decrease in erectile responses and increased apoptosis and fibrosis compared to these in the S group. Meanwhile, nerve content and the penile atrophy index were also decreased in the M group. Treatment with salidroside and rapamycin for 3weeks partially restored erectile function and significantly attenuated corporal apoptosis, fibrosis, nerve content and penile atrophy in the M group. Moreover, the autophagy level was further enhanced in the M+salidroside group, which was the same as that in the positive observation group (M+rapamycin). Salidroside treatment not only improved erectile function in rats with BCNI, but also inhibited apoptosis and fibrosis and ameliorated the loss of nerve content and endothelial and corpus cavernosum smooth muscle cells by promoting protective autophagy.

Local continuous glial cell derived neurotrophic factor release using osmotic pump promotes parasympathetic nerve rehabilitation in an animal model of cavernous nerve injury induced erectile dysfunction.

BackgroundNerve injury-related erectile dysfunction (ED) is one of the types that respond poorly to conventional ED treatments. Our previous experiments have demonstrated the paracrine of various neurotrophic factors (NTFs) by stem cells or other treatment modalities as a potential mechanism in the recovery of nerve injury-related ED. Glial cell derived neurotrophic factor (GDNF) is one of the essential NTFs for the regeneration of nerve fibers, especially for parasympathetic nerves. The aim of this study is to explore if local continuous GDNF administration is beneficial for the functional and histological recovery of nerve injury induced ED.MethodsEight-week-old male Sprague-Dawley rats were used for this study. Rats were randomly grouped into 5: Sham surgery (Sham), bilateral cavernous nerve injury (BCNI) and placebo treatment, BCNI and 0.1 µg/100 µL GDNF treatment (BCNI+GDNF 0.1), BCNI and 1 µg/100 µL GDNF treatment (BCNI+GDNF 1), BCNI and 10 µg/100 µL GDNF treatment (BCNI+GDNF 10). GDNF was administered using an osmotic pump technique which would deliver GDNF locally and continuously for 28 days without the need for external connections or frequent handling of animals. Recovery of sexual function, nerve fibers regeneration, and expression of neurotrophic receptors were examined and compared among groups after the treatment.ResultsLocal continuous GDNF release treatment increased the average number of intromissions in the sexual behavior test and intracavernous pressure (ICP) in the erectile function test in a dose dependent manner. Osmotic pump implantation induced increased local GDNF concentration and mild inflammatory response. Gene expression of GDNF receptors in major pelvic ganglion (MPG) and nerve regeneration along the urethra were partially promoted by GDNF. These changes were associated with increased nerve fibers especially the parasympathetic nerve fibers in dorsal nerve of penis (DNP) in GDNF treated groups.ConclusionsIn conclusion, our project illustrated the promising effects of local continuous GDNF administration for the functional and histological recovery of nerve injury-induced ED.

Administration of H2S improves erectile dysfunction by inhibiting phenotypic modulation of corpus cavernosum smooth muscle in bilateral cavernous nerve injury rats

Phenotypic modulation of Corpus Cavernosum Smooth Muscle Cells (CCSMCs) is an important step in the development and progression of bilateral cavernous nerve injury induced erectile dysfunction (BCNI-ED). To investigate the effect of exogenous hydrogen sulfide (H2S) on the phenotypic modulation of CCSMCs in BCNI-ED rats, a total of 18 male Sprague-Dawley rats were equally divided into 3 groups, including sham-operated (Sham) group, BCNI group and BCNI treated with NaHS (BCNI + NaHS) group. The treated group received intraperitoneal injection of NaHS (100 μmol kg−1day−1) for 4 weeks starting day 1 postoperatively. Erectile function was measured by the ratio of intracavernous pressure (ICP)/mean arterial pressure (MAP), and relevant tissues were harvested for Immunohistochemistry, Hematoxylin and eosin (H&E), Masson's trichrome staining, H2S fluorescent probe WSP-1 and Western blot. The primary CCSMCs were isolated and pretreatment with NaHS before exposed to PDGF-BB (platelet-derived growth factor). Relative expression mRNA and protein of phenotypic biomarkers, RhoA, ROCK-1 and cell cycle proteins were detected. Cystathionine-β-synthase (CBS) and cystathionine-γ-lyase (CSE), 3-mercaptopyruvate sulfurtransferase (3-MST) and H2S levels in penile tissue was significantly decreased in the BCNI group compared with the Sham group. Compared with the BCNI group, administration of NaHS significantly increased the ratio of ICP/MAP, ratio of smooth muscle to collagen, expressions of a-SMA, calponin and decreased the expression of OPN, collagen-I, RhoA, ROCK1 in the penile tissue. PDGF-BB-treated CCSMCs exhibited higher expression of OPN, RhoA, ROCK1, and lower α-SMA, calponin, which were attenuated by NaHS pretreatment. NaHS suppressed RhoA/ROCK activity and decreased the expression of CDK2, Cyclin E1, while increased the expression of P27kip1 induced by PDGF-BB in CCSMCs. Taken together, this study indicated that exogenous H2S inhibited the phenotypic modulation of CCSMCs by suppressing RhoA/ROCK1 signaling and affecting its downstream factor, CDK2, Cyclin E1, P27kip1, thereby improved BCNI rat erectile function.

Oral Administration of the p75 Neurotrophin Receptor Modulator, LM11A-31, Improves Erectile Function in a Mouse Model of Cavernous Nerve Injury

BackgroundRadical prostatectomy for prostate cancer can not only induce cavernous nerve injury (CNI), but also causes cavernous hypoxia and cavernous structural changes, which lead to a poor response to phosphodiesterase 5 inhibitors. AimTo investigate the therapeutic effect of oral administration of LM11A-31, a small molecule p75 neurotrophin receptor (p75NTR) ligand and proNGF antagonist, in a mouse model of bilateral CNI, which mimics nerve injury–induced erectile dysfunction after radical prostatectomy. Methods8-week-old male C57BL/6 mice were divided into sham operation and CNI groups. Each group was divided into 2 subgroups: phosphate-buffered saline and LM11A-31 (50 mg/kg/day) being administered once daily starting 3 days before CNI via oral gavage. 2 weeks after CNI, we measured erectile function by electrical stimulation of the bilateral cavernous nerve. The penis was harvested for histologic examination and Western blot analysis. The major pelvic ganglia was harvested and cultured for assays of ex vivo neurite outgrowth. OutcomesIntracavernous pressure, neurovascular regeneration in the penis, in vivo or ex vivo functional evaluation, and cell survival signaling were measured. ResultsErectile function was decreased in the CNI group (44% of the sham operation group), while administration of LM11A-31 led to a significant improvement of erectile function (70% of the sham operation group) in association with increased neurovascular content, including cavernous endothelial cells, pericytes, and neuronal processes. Immunohistochemical and Western blot analyses showed significantly increased p75NTR expression in the dorsal nerve of CNI mice, which was attenuated by LM11A-31 treatment. Protein expression of active PI3K, AKT, and endothelial nitric oxide synthase was increased, and cell death and c-Jun N-terminal kinase signaling was significantly attenuated after LM11A-31 treatment. Furthermore, LM11A-31 promoted neurite sprouting in cultured major pelvic ganglia after lipopolysaccharide exposure. Clinical ImplicationsLM11A-31 may be used as a strategy to treat erectile dysfunction after radical prostatectomy or in men with neurovascular diseases. Strengths & LimitationsUnlike biological therapeutics, such as proteins, gene therapies, or stem cells, the clinical application of LM11A-31 would likely be relatively less complex and low cost. Our study has some limitations. Future studies will assess the optimal dosing and duration of the compound. Given its plasma half-life of approximately 1 hour, it is possible that dosing more than once per day will provide added efficacy. ConclusionSpecific inhibition of the proNGF-p75NTR degenerative signaling via oral administration of LM11A-31 represents a novel therapeutic strategy for erectile dysfunction induced by nerve injury.Yin GN, Ock J, Limanjaya A, et al. Oral Administration of the p75 Neurotrophin Receptor Modulator, LM11A-31, Improves Erectile Function in a Mouse Model of Cavernous Nerve Injury. J Sex Med 2021;18:17–28.

Implantation of Skin-Derived Precursors Derived Schwann Cells Improves Erectile Function in a Bilateral Cavernous Nerve Injury Rat Model

Objectives: In this study, we used skin-derived precursors derived Schwann cells (SKP-SCs) as a treatment to alleviate the erectile dysfunction resulting from bilateral cavernous nerve injury (BCNI) in rat models. And the effect was evaluated 2 weeks after treatment. Materials and Methods: SKP-SCs and primary Schwann cells(SCs) were extracted and incubated from 4-week-old male Sprague Dawley(SD) rats. The cells were used to co-culture with PC12 cells for measurement of neurites growth. Conditional medium of them was used for chemotaxis of M0 macrophages and ELISA assays. 24 male SD rats (10-week old) were randomly divided into 4 groups (n=6). One group was sham group and the other 3 groups underwent BCNI. The groups undergoing BCNI were treated with PBS, SKP-SCs, primary SCs respectively. We evaluated the therapeutic effects on BCNI rats by measuring the erectile function, histologic analysis and western blotting of penis and MPG tissues. Results: Treatment with SKP-SCs significantly restored the erectile function, accelerated the recovery of endothelial and smooth muscle tissues in the penis and promoted the nerve repair process. Expression of p-smad2 and p-smad3 decreased after treatment, indicating fibrosis in the corpus cavernosum was significantly blocked. Conclusion: Implantation of SKP-SCs is an effective and promising therapy for erectile dysfunction induced by BCNI. Funding Statement: Support of research grants from National Natural Science Foundation of China (No. 81971378) and Science and Technology Program of Guangzhou (No. 201903010027). Declaration of Interests: We claim that no support or financial relationships have been received from any organization that may have an interest in the submitted work, and there are no other relationships or activities that could appear to have influenced the submitted work. Ethics Approval Statement: The present study has been reviewed and permitted by the Institutional Animal Care and Use Subcommittee of the Third Affiliated Hospital of Sun Yat-sen University.

Neutralizing antibody to proNGF rescues erectile function by regulating the expression of neurotrophic and angiogenic factors in a mouse model of cavernous nerve injury.

Radical prostatectomy induces some degree of cavernous nerve injury (CNI) and causes denervation-induced pathologic changes in cavernous vasculature, regardless of the advances in surgical techniques and robotic procedures. The precursor for nerve growth factor (proNGF) is known to be involved in neuronal cell apoptosis and microvascular dysfunction through its receptor p75NTR . To determine the expression of proNGF/p75NTR and the efficacy of proNGF neutralizing antibody (anti-proNGF-Ab) in a mouse model of ED induced by CNI. Age-matched 12-week-old C57BL/6 mice were distributed into three groups: sham group and bilateral CNI group treated with intracavernous injections of PBS (20μL) or of anti-proNGF-Ab (20µg in 20μL of PBS) on days -3 and 0. Two weeks after treatment, erectile function was measured by electrical stimulation of cavernous nerve. Penis tissues from a separate group of animals were harvested for further analysis. We also determined the efficacy of anti-proNGF-Ab on neural preservation in major pelvic ganglion (MPG) ex vivo. We observed increased penile expression of proNGF and p75NTR after CNI. Intracavernous administration of anti-proNGF-Ab increased nNOS and neurofilament expression probably by enhancing the production of neurotrophic factors, such as neurotrophin-3, NGF, and brain-derived neurotrophic factor. Anti-proNGF-Ab preserved the integrity of cavernous sinusoids, such as pericytes, endothelial cells, and endothelial cell-to-cell junctions, possibly by controlling angiogenic factors (angiopoietin-1, angiopoietin-2, and vascular endothelial growth factor) and induced endogenous eNOS phosphorylation in CNI mice. And finally, treatment with anti-proNGF-Ab rescued erectile function in CNI mice. Anti-proNGF-Ab also enhanced neurite sprouting from MPG exposed to lipopolysaccharide. The preservation of damaged cavernous neurovasculature through inhibition of the proNGF/p75NTR pathway may be a novel strategy to treat radical prostatectomy-induced erectile dysfunction.

Neuroprotective effect of Hongjing I granules on erectile dysfunction in a rat model of bilateral cavernous nerve injury

Neurogenic erectile dysfunction (NED) is an inevitable postoperative disease of cavernous nerve injury which will lead to various pathophysiological changes in the corpus cavernosum and dorsal penile nerve caused by radical prostatectomy (RP). Although serval years of clinical application of HJIG I granules (HJIG), an innovative formulation, has demonstrated its reliable clinical efficacy against NED, the mechanism of HJIG remains unclear. This study aimed to assess the neuroprotective effect of HJIG, to repair damaged nerves in a rat model of bilateral cavernous nerve injury (BCNI) in vivo and their effects on neurites of major pelvic ganglia (MPG) regeneration and Schwann cells (SCs) proliferation in vitro. Rats were divided into five groups randomly: normal control (NC), BCNI-induced ED model (M), M + low-dose HJIG (HL), M + medium-dose HJIG (HM), and M + high-dose HJIG (HH). All groups were treated with normal saline or the relevant drug for 28 consecutive days after a standard NED animal model. Our data revealed that administration of HJIG improved NED that was detected by intracavernous pressure (ICP) in a dose-dependent manner. The haematoxylin-eosin (HE) and Immunofluorescence (IF) staining demonstrated that HJIG ameliorate the shape of penis and induced the protein synthesis of GAP43, NF200, S100, and nNOS. NF200 and S100 level were also detected by western blotting. Moreover, HJIG (0.78 mg/mL) markedly increased SCs viability and promoted neurites regeneration of MPG. These findings provide new insights into the NED therapy by HJIG.

HP-1-4 Filtrated Bone Marrow-Derived Mesenchymal Stem Cell Lysate Improves Erectile Function in a Rat Model of Cavernous Nerve Injury

Oral phosphodiesterase-5 (PDE5) inhibitors and intracavernous injection of prostaglandin E1 (PGE1) are treatments of choice for post radical prostatectomy (RP) erectile dysfunction (ED). However, these treatments are not necessarily effective and the development of a novel and effective therapy for post RP ED is required. Recently, it was reported that mesenchymal stem cell (MSC) transplantation induces nerve regeneration and improves erectile function in a rat model of bilateral cavernous nerve injury (BCNI). However, unexpected differentiation of the transplanted stem cells is still an issue with this approach. To avert this problem, we explored the suitability of filtrated stem cell lysate (FSCL), which contains secreted factors but does not contain MSCs. In this study, we investigated the effects of FSCL on erectile function in a rat model of BCNI. Eight-week-old male Wistar/ST rats underwent BCNI or sham surgery. After surgery, filtrated adipose-derived stem cell lysate (FADSCL) or filtrated bone marrow-derived stem cell lysate (FBMSCL) was injected into the corpus cavernosum. After 4 weeks, erectile function was evaluated by measuring intracavernosal pressure/mean arterial pressure under stimulation of the cavernous nerve. Proteomic analysis was performed to identify effective factors in FSCL.

MP84-08 TUMOR NECROSIS FACTOR ALPHA IS INCREASED AFTER CAVERNOUS NERVE INJURY AND IMPAIRS REGENERATION OF NITRERGIC NEURONS AND MODULATES PENILE SMOOTH MUSCLE TONE

You have accessJournal of UrologySexual Function/Dysfunction: Basic Research & Pathophysiology (MP84)1 Apr 2020MP84-08 TUMOR NECROSIS FACTOR ALPHA IS INCREASED AFTER CAVERNOUS NERVE INJURY AND IMPAIRS REGENERATION OF NITRERGIC NEURONS AND MODULATES PENILE SMOOTH MUSCLE TONE Hotaka Matsui*, Nikolai A. Sopko, Jefferey D. Campbell, Xiaopu Liu, Emmanuel Weyne, Maarten Albersen, Fabio Castiglione, Johanna L. Hannan, and Bivalacqua Trinity J Hotaka Matsui*Hotaka Matsui* More articles by this author , Nikolai A. SopkoNikolai A. Sopko More articles by this author , Jefferey D. CampbellJefferey D. Campbell More articles by this author , Xiaopu LiuXiaopu Liu More articles by this author , Emmanuel WeyneEmmanuel Weyne More articles by this author , Maarten AlbersenMaarten Albersen More articles by this author , Fabio CastiglioneFabio Castiglione More articles by this author , Johanna L. HannanJohanna L. Hannan More articles by this author , and Bivalacqua Trinity JBivalacqua Trinity J More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000000976.08AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Erectile dysfunction is induced after cavernous nerve (CN) injury by inflammation in pelvic ganglia. We examined levels of tumor necrosis factor alpha (TNFA), after bilateral CN injury (BCNI) in vivo, and effect of exogenous TNFA (eTNFA) on neurite outgrowth from major pelvic ganglion (MPG) ex vivo. Additionally, we evaluated effect of TNFA signal inhibition to penile smooth muscle tone with TNFA receptor knockout mice (TNFRKO). METHODS: 70 male rats underwent sham or BCNI. MPGs of sham rats were collected after 48 hours. BCNI groups had MPG harvested at 6 h, 12 h, 24 h, 48 h, 7 days, or 14 days after surgery. We performed qPCR to examine gene expression of TNFA and Western blot to examine protein amount of TNFA in MPGs. Immunofluorescence(IF) for TNFA was performed in MPG. For ex vivo study, MPGs were harvested from control rats and cultured in Matrigel with eTNFA (0, 10, 20, 30 ng/mL). Neurite lengths from MPGs were measured 72 h after culture. MPGs were processed for qPCR of Tyrosine hydroxylase (TH) and neuronal nitric oxidase (nNOS). IF was assessed for TH and nNOS in MPG. To examine effect of TNFA inhibition on MPG and penis after BCNI, wild type mice (WT) and TNFRKO were randomized to undergo sham or BCNI. MPGs were harvested 48 hours after surgery to examine gene expression of nNOS and TH by qPCR. Penises were used to examine smooth muscle tone to electrical field stimulation (EFS). RESULTS: TNFA’s gene/protein expression was increased in MPGs after BCNI, with peaks at 2 and 7 days, respectively. IF showed TNFA localized to perivascular areas at 48 h and neuronal cell body at 14 days after BCNI. eTNFA inhibited neurite outgrowth in a dose-dependent fashion, and impaired gene expression of nNOS in MPG (p<0.05). MPGs cultured with eTNFA showed significantly shorter neurite lengths of nNOS+ neurons than TH+ neurons (p<0.01), while this difference was not seen in controls. TNFRKO showed enhanced gene expression of nNOS compared to WT (p<0.01). Penises of WT showed enhanced smooth-muscle contraction to EFS, compared to TNFKO (p<0.01). TNFRKO had enhanced relaxation response to EFS after maximal contraction, compared to WT (p<0.01). CONCLUSIONS: TNFA was increased following BCNI and inhibited neurite outgrowth from MPGs in a dose-dependent fashion. eTNFA lowered gene and protein expression of nNOS. TNFRKO mice showed enhanced gene expression of nNOS and enhanced penile smooth muscle relaxation. These results suggest that TNFA may selectively inhibit nNOS+ neurons, and impairs penile smooth muscle tone. Source of Funding: NIH © 2020 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 203Issue Supplement 4April 2020Page: e1267-e1267 Advertisement Copyright & Permissions© 2020 by American Urological Association Education and Research, Inc.MetricsAuthor Information Hotaka Matsui* More articles by this author Nikolai A. Sopko More articles by this author Jefferey D. Campbell More articles by this author Xiaopu Liu More articles by this author Emmanuel Weyne More articles by this author Maarten Albersen More articles by this author Fabio Castiglione More articles by this author Johanna L. Hannan More articles by this author Bivalacqua Trinity J More articles by this author Expand All Advertisement PDF downloadLoading ...

Stem Cell/Oxygen-Releasing Microparticle Enhances Erectile Function in a Cavernous Nerve Injury Model.

Erectile dysfunction caused by damage to the cavernous nerve is a common complication of radical prostatectomy for patients with localized prostate cancer. Various studies have investigated repair of damaged tissue and prevention of fibrosis in the corpus cavernosum using stem cell therapy. However, stem cell therapy has limitations, including insufficient nutrient and oxygen supply to transplanted stem cells. This study investigated whether stem cell/oxygen-releasing hollow microparticles (HPs) had therapeutic effect on erectile dysfunction in a rat model of bilateral cavernous nerve injury (BCNI). Therapeutic effects were observed in the BCNI model at 1, 2, and 4 weeks postcavernous nerve injury. Erectile function further improved after treatment with stem cell/oxygen-releasing HP system compared with treatment with only stem cells at 4 weeks. Stem cell/oxygen-releasing HP system increased cyclic guanosine monophosphate (cGMP) level and neuronal nitric oxide synthase (nNOS), endothelial nitric oxide synthase (eNOS), α-smooth muscle actin (α-SMA), and muscarinic acetylcholine receptor 3 (M3) expression while decreasing fibrosis and apoptosis in the corpus cavernosum. Our results clearly show that stem cell survival increases around transplanted stem cell/oxygen-releasing hybrid system site. Taken together, an oxygen-releasing HP system supported prolonged stem cell survival, sustaining the paracrine effect of the stem cells, and consequently enhancing erectile function. These findings show promise with regard to prolonged stem cell survival in stem cell applications for various diseases and types of tissue damage. Impact statement In this study, we used an oxygen-releasing hollow microparticles (HPs) system with stem cells to attempt to overcome certain limitations of stem cell therapy, including insufficient nutrient and oxygen supplies for transplanted stem cells. Our results demonstrated that a stem cell/oxygen-releasing HP hybrid system could further improve erectile function, cyclic guanosine monophosphate (cGMP) level, and NOS level in a bilateral cavernous nerve injury rat model through prolonged stem cell survival. Our data suggest that a stem cell/oxygen-releasing HP system is a promising clinical treatment option for postprostatectomy erectile dysfunction. Furthermore, this system may be relevant in different disease therapies and regenerative medicine.

Icariside II facilitates the differentiation of ADSCs to schwann cells and restores erectile dysfunction through regulation of miR-33/GDNF axis

BackgroundAdipose derived stem cells (ADSCs) have the property to differentiate into neuron-like cells, which may provide a novel insight for the restoration of erectile dysfunction (ED) mainly induced by cavernous nerve injury. Icariside II (ICA II) has been reported to play a key role in the regulation of erectile function via stimulating the differentiation of ADSCs to Schwann Cells (SCs). However, the function and molecular mechanisms of ICA II in ED remains to be further clarified. MethodsThe expression of S100, P75, GDNF and miR-33 was detected by qRT-PCR. And the relative proteins expression was determined by western blot. Cell viability was measured by Cell Counting Kit-8 (CCK-8) assay. Bioinformatics, luciferase reporter and RNA immunoprecipitation (RIP) assays were performed to verify the interaction between miR-33 and GDNF. Intracavernosal pressure (ICP), the ratio of ICP and mean arterial pressure (MAP), as well as nNOS expression were examined to evaluate the erectile function of SD rats with bilateral cavernous nerve injury (BCNI). ResultsICA II and miR-33 respectively promoted and inhibited the differentiation of ADSCs to SCs. MiR-33 could negatively regulate P75 and GDNF expression. ICA II exerted promotion effects on differentiation of ADSCs to SCs via regulating miR-33. GDNF was identified to be a target of miR-33. MiR-33 overexpression abrogated the stimulatory effect of ICA II on ADSCs' differentiation, which was blocked by GDNF overexpression. treated with ICA II recovered the erectile function of BCNI model rats through regulation of miR-33. ConclusionICA II contributed to the differentiation of ADSCs to SCs viamiR-33/GDNF axis, contributing to the recovery of erectile function in BCNI rats.

Co-overexpression of VEGF and GDNF in adipose-derived stem cells optimizes therapeutic effect in neurogenic erectile dysfunction model.

ObjectivesTo evaluate the rapid repair potential of adipose‐derived stem cells (ADSCs) co‐overexpressing VEGF and GDNF on bilateral cavernous nerve injury (BCNI) in rat models. Progressive fibrosis of the penis that occurs shortly after BCNI is a key cause of clinical treatment difficulty of erectile dysfunction (ED). Traditional medications are ineffective for ED caused by BCNI. ADSCs have shown therapeutic effects in animal models, but disappointing in clinical treatment suggests that we should explore optimal treatment of it.Materials and methodsWe extracted ADSCs from rat epididymis. Lentiviral transfection was verified by western blot and immunofluorescence. Thirty‐six SD rats (10 weeks old) were randomly divided into six groups (n = 6 per group): sham surgery, and remaining five BCNI groups transplanted PBS or ADSCs which were genetically modified by vehicle, VEGF (ADSC‐V), GDNF (ADSC‐G), or VEGF&GDNF (ADSC‐G&V) around major pelvic ganglion (MPG). We investigated the therapeutic effects of BCNI rat model which is characterized by ED, penile tissue fibrosis and hypoxia, and lack of nitrogen nerves or vascular atrophy.ResultsErectile function was almost recovered after 2 weeks of transplantation of ADSC‐G&V, promoted cavernous nerve repair, prevented penile fibrosis and preserving the vascular endothelium, which was significant differences amongst ADSC‐V or ADSC‐G. Moreover, GM‐ADSCs were detected in MPG and penis, indicating that their participation in repair of target organs and transverse nerves.ConclusionsThese promising data indicate that ADSCs co‐overexpressed VEGF and GDNF‐induced synergistic effects, make it a potential tool for recovering of erectile function speedily after BCNI.

171 Increased Level of Tumor Necrosis Factor-alpha (TNFA) Leads to Downregulation of Nitrergic Neurons Following Bilateral Cavernous Nerve Injury and TNFA Inhibition Can Enhance Penile Smooth Muscle Relaxation by Upregulating Expression of Nitrergic Neurons

171 Increased Level of Tumor Necrosis Factor-alpha (TNFA) Leads to Downregulation of Nitrergic Neurons Following Bilateral Cavernous Nerve Injury and TNFA Inhibition Can Enhance Penile Smooth Muscle Relaxation by Upregulating Expression of Nitrergic Neurons

Exogenous hydrogen sulfide improves erectile dysfunction by inhibiting apoptosis of corpus cavernosum smooth muscle cells in rats with cavernous nerve injury

To explore the effects of exogenous hydrogen sulfide (H2S) on apoptosis of corpus cavernosum smooth muscle cells (CCSMCs) and erectile dysfunction (ED) in rats with bilateral cavernous nerve injury (BCNI). Twentyfour male SD rats were randomly divided into 3 groups (n=8):sham operation group, bilateral cavernous nerve injury group (BCNI group) and H2S intervention group (BCNI+NaHS group). In BCNI and BCNI+NaHS groups, BCNI was induced by clamp injury of the bilateral cavernous nerves, and the rats were subjected to daily intraperitoneal injection of normal saline and 100 μmol/kg NaHS solution for 4 weeks, respectively. After the treatment, the intracavernous pressure (ICP) and mean arterial pressure (MAP), ) of the rats were measured. Western blotting was used to detect the expressions of cystathionine β synthetase (CBS), cystathionine γ lyase (CSE), α-SMA, collagen-I, caspase-3, Bax and Bcl-2 in the penile cavernous tissue, and the expressions of CBS and CSE were also detected immunohistochemically. The ratio of cavernous smooth muscle to collagen was detected using Masson's Trichrome staining. The apoptosis level of CCSMC was detected by TUNEL + α-SMA immunofluorescence double staining. After 4 weeks of treatment, the rats in BCNI+NaHS group showed a significantly higher ICP/MAP ratio than those in BCNI group (P < 0.05). The results of Masson's Trichrome staining showed that the ratio of cavernous smooth muscle/collagen was significantly higher in BCNI + NaHS group than in BCNI group (P < 0.05). Western blotting showed a significantly higher expression of α-SMA protein but a lower expression of collagen-I protein in BCNI + NaHS group than in BCNI group (P < 0.05). TUNEL+α-SMA immunofluorescence double staining revealed a significantly lower number of apoptotic CCSMCs in BCNI+NaHS group than in BCNI group (P < 0.05). Compared with those in BCNI group, the rats in BCNI+NaHS group had significantly decreased expressions of caspase-3 and Bax proteins (P < 0.05) with significantly enhanced Bcl-2 protein expression and an increased Bcl-2/Bax ratio (P < 0.05). The expressions of CBS and CSE were significantly lower in BCNI group than in the other two groups (P < 0.05). Exogenous H2S enhance the expression of the classic apoptotic protein Bcl-2 and reduces apoptosis of CCSMC to improve the erectile function in rats with BCNI.

Smooth Muscle Progenitor Cells Preserve the Erectile Function by Reducing Corporal Smooth Muscle Cell Apoptosis after Bilateral Cavernous Nerve Crush Injury in Rats.

Radical prostatectomy causes erectile dysfunction (ED) and irreversible morphologic changes, including induction of endothelial and smooth muscle cell (SMC) apoptosis in the corpus cavernosum (CC). The injection of smooth muscle progenitor cells (SPCs) thickens the vascular intima and has demonstrated therapeutic benefit in cardiovascular disease animal. Herein, we investigated the effect of SPCs on the recovery of erectile function (EF) in rat models with bilateral cavernous nerve (CN) injury. Twenty-four male Sprague-Dawley rats were randomized into sham, vehicle only, or SPC treatment groups. Rats in the SPC treatment and vehicle groups were subjected to bilateral CN injury before intracavernosal injection. Intracavernosal injections of SPCs increased all EF parameters at day 28 after injury and simultaneously reduced apoptosis of the SMCs. Ultrastructural analysis revealed that SPCs maintained the integrity of the CC by preserving the structure of the adherens junctions. Tracking transplanted SPCs labeled with EdU showed that transplanted SPCs remained in the CC 28 days after treatment. Intracavernosal SPC injection restored EF after bilateral CN injury by reducing SMC apoptosis, which favored the maintenance of the structure of adherens junctions and regulated the stability of corporal vessels. These findings demonstrate the therapeutic potential of SPCs for treating ED in humans.