ObjectiveTo investigate the protective effect of human urine-derived stem cells (USCs) on erectile function and cavernous structure in rats with cavernous nerve injury (CNI).

METHODSSixty adult male SD rats with normal sexual function were randomly divided into four groups of equal number: sham operation, bilateral CNI (BCNI) model control, phosphate buffered saline (PBS), and USC. The BCNI model was established in the latter three groups of rats by clamping the bilateral cavernous nerves. After modeling, the rats in the PBS and USC groups were treated by intracavernous injection of PBS at 200 μl and USCs at 1×106/200 μl PBS respectively for 28 days. Then, the maximum intracavernous pressure (mICP) and the ratio of mICP to mean arterial pressure (mICP/MAP) of the rats were calculated by electrical stimulation of the major pelvic ganglions, the proportion of nNOS- or NF200-positive nerve fibers in the total area of penile dorsal nerves determined by immunohistochemical staining, the levels of endothelial cell marker eNOS, smooth muscle marker α-SMA and collagen I detected by Western blot, and the smooth muscle to collagen ratio and the cell apoptosis rate in the corpus cavernosum measured by Masson staining and TUNEL, respectively.

RESULTSAfter 28 days of treatment, the rats in the USC group, as compared with those in the PBS and BCNI model control groups, showed significant increases in the mICP (［81 ± 9.9］ vs ［31 ± 8.3］ and ［33 ± 4.2］ mmHg, P <0.05), mICP/MAP ratio (0.72 ± 0.05 vs 0.36 ± 0.03 and 0.35 ± 0.04, P <0.05), the proportions of nNOS-positive nerve fibers (［11.31 ± 4.22］% vs ［6.86 ± 3.08］% and ［7.29 ± 4.84］% , P <0.05) and NF200-positive nerve fibers in the total area of penile dorsal nerves (［27.31 ± 3.12］% vs ［17.38 ± 2.87］% and ［19.49 ± 4.92］%, P <0.05), the eNOS/GAPDH ratio (0.52 ± 0.08 vs 0.31 ± 0.06 and 0.33 ± 0.07, P <0.05), and the α-SMA/GAPDH ratio (1.01 ± 0.09 vs 0.36 ± 0.05 and 0.38 ± 0.04, P <0.05), but a remarkable decrease in the collagen I/GAPDH ratio (0.28 ± 0.06 vs 0.68 ± 0.04 and 0.70 ± 0.10, P <0.05). The ratio of smooth muscle to collagen in the corpus cavernosum was significantly higher in the USC than in the PBS and BCNI model control groups (17.91 ± 2.86 vs 7.70 ± 3.12 and 8.21 ± 3.83, P <0.05) while the rate of cell apoptosis markedly lower in the former than in the latter two (3.31 ± 0.83 vs 9.82 ± 0.76, P <0.01; 3.31 ± 0.83 vs 9.75 ± 0.91, P <0.05).

CONCLUSIONSIntracavernous injection of USCs can protect the erectile function of the rat with cavernous nerve injury by protecting the nerves, improving the endothelial function, alleviating fibrosis and inhibiting cell apoptosis in the cavernous tissue.

Actins ; analysis ; Animals ; Arterial Pressure ; Collagen ; analysis ; Disease Models, Animal ; Erectile Dysfunction ; prevention & control ; Male ; Nitric Oxide Synthase Type I ; analysis ; Nitric Oxide Synthase Type III ; analysis ; Penile Erection ; physiology ; Penis ; innervation ; Pudendal Nerve ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Saline Solution ; administration & dosage ; Stem Cell Transplantation ; methods ; Stem Cells ; Urine ; cytology

We evaluated whether LIM-kinase 2 inhibitor (LIMK2i) could improve erectile function by suppressing corporal fibrosis through the normalization of the Rho-associated coiled-coil protein kinase 1 (ROCK1)/LIMK2/Cofilin pathway in a rat model of cavernous nerve crush injury (CNCI). Sixty 11-week-old male Sprague-Dawley rats were divided equally into five groups: sham surgery (S), CNCI (I), and CNCI treated with low-dose (L), medium-dose (M), and high-dose (H) LIMK2i. The L, M, and H groups were treated with a daily intraperitoneal injection of LIMK2i (2.5, 5.0, and 10.0 mg kg^-1 body weight, respectively) for 1 week after surgery. The erectile response was assessed using electrostimulation at 1 week, postoperatively. Penile tissues were processed for Masson's trichrome staining, double immunofluorescence, and Western blot assay. Erectile responses in the H group improved compared with the I group, while the M group showed only partial improvement. A significantly decreased smooth muscle/collagen ratio and an increased content of fibroblasts positive for phospho-LIMK2 were noted in the I group. The M and H groups revealed significant improvements in histological alterations and the dysregulated LIMK2/Cofilin pathway, except for LIMK2 phosphorylation in the M group. The inhibition of LIMK2 did not affect the ROCK1 protein expression. The content of fibroblasts positive for phospho-LIMK2 in the H group returned to the level found in the S group, whereas it did not in the M group. However, the L group did not exhibit such improvements. Our data suggest that the inhibition of LIMK2, particularly with administration of 10.0 mg kg^-1 body weight LIMK2i, can improve corporal fibrosis and erectile function by normalizing the LIMK2/Cofilin pathway.
Animals ; Cofilin 1/metabolism* ; Electric Stimulation ; Erectile Dysfunction/etiology* ; Fibroblasts/pathology* ; Fibrosis/drug therapy* ; Lim Kinases/antagonists & inhibitors* ; Male ; Penile Diseases/drug therapy* ; Penis/innervation* ; Peripheral Nerve Injuries/pathology* ; Phosphorylation ; Rats ; Rats, Sprague-Dawley ; Signal Transduction/drug effects* ; rho-Associated Kinases/genetics*

The nerve electrophysiological tests may differentiate the treatment of primary premature ejaculation (PPE) in our previous studies. However, no study verifies if the results will be affected by abstinence time. From January to December in 2016, fifty PPE patients ejaculated within 2 min and 28 control subjects were enrolled. The nerve electrophysiological tests, including dorsal nerve somatosensory evoked potential (DNSEP), glans penis somatosensory evoked potential (GPSEP), and penile sympathetic skin response (PSSR), were recorded before and immediately after ejaculation. The abstinence day was not correlated with the latencies of SEPs or PSSR neither in PE group (P = 0.170, 0.064, and 0.122, respectively) nor in control group (P = 0.996, 0.475, and 0.904, respectively). No statistically differences were found in the latencies of SEPs and PSSR before and after ejaculation in PE patients (P = 0.439, 0.537, and 0.576, respectively) or control subjects (P = 0.102, 0.198, and 0.363, respectively). Thus, abstinence time does not interfere with the nerve electrophysiological test, which is stable in determining the nerve function of PPE patients.
Adult ; Ejaculation ; Electric Stimulation ; Electrophysiological Phenomena ; Evoked Potentials, Somatosensory ; Humans ; Male ; Middle Aged ; Penis/physiopathology* ; Premature Ejaculation/physiopathology* ; Prospective Studies ; Sexual Abstinence ; Skin/innervation* ; Sympathetic Nervous System/physiopathology* ; Young Adult

Stem cell therapy is a potentially promising option for erectile dysfunction; however, its risk of tumorigenicity is a clinical hurdle and the risk is positively related to the number of injected cells. Our previous study showed that nanotechnology improved adipose-derived stem cell (ADSC) therapy for erectile dysfunction of cavernous nerve injury (CNI) by attracting cells in the corpus cavernosum. These results indicated the possibility of using a reduced dosage of ADSCs for intracavernous injection. In this exploratory study, we used lower dosage (2 × 10⁵ cells) of ADSCs for intracavernous injection (ICI) and the nanotechnology approach. Intracavernous pressure and mean arterial pressure were measured at day 28 to assess erectile function. The low-dose ADSC therapy group showed favorable treatment effects, and nanotechnology further improved these effects. In vivo imaging of ICI cells revealed that the fluorescein signals of NanoShuttle-bound ADSCs (NanoADSCs) were much stronger than those of ADSCs at days 0, 1, and 3. Both immunofluorescence and Western blot analysis showed a significant increase in smooth muscle, endothelium, and nerve tissue in the ADSC group compared to that in the CNI group; further improvement was achieved with assisted nanotechnology. These findings demonstrate that nanotechnology can be used to further improve the effect of small dosage of ADSCs to improve erectile function. Abundant NanoADSCs remain in the corpus cavernosum in vivo for at least 3 days. The mechanism of erectile function improvement may be related to the regeneration of the smooth muscle, endothelium, and nerve tissues.
Animals ; Cell Tracking ; Disease Models, Animal ; Erectile Dysfunction/therapy* ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stem Cells ; Penis/innervation* ; Peripheral Nerve Injuries/complications* ; Rats ; Rats, Sprague-Dawley ; Treatment Outcome

Objective: To explore the effects of immediate and delayed intracavernous injection of bone marrow mesenchymal stem cells (BM-MSCs) on neurogenic erectile dysfunction (NED) induced by bilateral cavernous nerve injury in Sprague-Dawley (SD) rats. METHODS: BM-MSCs isolated from male SD rats were cultured and identified. Twenty-eight 8-week-old male SD rats were randomly divided into four groups, sham operation, NED model control, BM-MSCs immediate, and BM-MSCs delayed, and NED models were established in the latter three groups by crushing the bilateral cavernous nerves. The rats in the sham operation and model control groups were injected intracavernously with placebo while those in the latter two with BM-MSCs immediately or 2 weeks after modeling. At 12 weeks after operation, the penile function of the rats was assessed according to the penile intracavernous pressure (ICP), mean arterial pressure (MAP), and ICP/MAP ratio obtained from different groups of rats. Then, all the animals were sacrificed and the penile cavernosal tissue collected for histological analysis. RESULTS: At 12 weeks after modeling, both ICP and ICP/MAP were significantly increased in the BM-MSCs immediate and delayed groups as compared with those in the model control (P <0.05), and so were the ratio of smooth muscle to collagen (P <0.05) and the smooth muscle content in the corpus cavernosum (P <0.05), and the number of neurofilament (NF)-positive nerve fibers (P <0.05) and the expression of neuronal nitric oxide synthase (nNOS) in the dorsal nerves of the midshaft penis (P <0.05). CONCLUSIONS Intracavernous injection of BM-MSCs can improve erectile function in rats with bilateral cavernous nerve injury by elevating the smooth muscle-collagen ratio and smooth muscle content in the corpus cavernosum and thus preventing its fibrosis as well as by increasing the number of NF-positive nerve fibers and expression of nNOS in the penile dorsal nerves.
Animals ; Disease Models, Animal ; Erectile Dysfunction ; enzymology ; etiology ; therapy ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Muscle, Smooth ; Nitric Oxide Synthase Type I ; metabolism ; Penile Erection ; physiology ; Penis ; enzymology ; innervation ; Pudendal Nerve ; Random Allocation ; Rats ; Rats, Sprague-Dawley

PURPOSE: This study was performed to examine the treatment of erectile dysfunction by use of superparamagnetic iron oxide nanoparticles-labeled human mesenchymal stem cells (SPION-MSCs) transplanted into the cavernous nerve injured cavernosa of rats as monitored by molecular magnetic resonance imaging (MRI). MATERIALS AND METHODS: Eight-week-old male Sprague-Dawley rats were divided into three groups of 10 rats each: group 1, sham operation; group 2, cavernous nerve injury; group 3, SPION-MSC treatment after cavernous nerve injury. Immediately after the cavernous nerve injury in group 3, SPION-MSCs were injected into the cavernous nerve injured cavernosa. Serial T2-weighted MRI was done immediately after injection and at 2 and 4 weeks. Erectile response was assessed by cavernous nerve stimulation at 2 and 4 weeks. RESULTS: Prussian blue staining of SPION-MSCs revealed abundant uptake of SPION in the cytoplasm. After injection of 1x10(6) SPION-MSCs into the cavernosa of rats, T2-weighted MRI showed a clear hypointense signal induced by the injection. The presence of SPION in the corpora cavernosa was confirmed with Prussian blue staining. At 2 and 4 weeks, rats with cavernous nerve injury had significantly lower erectile function than did rats without cavernous nerve injury (p<0.05). The group transplanted with SPION-MSCs showed higher erectile function than did the group without SPION-MSCs (p<0.05). The presence of SPION-MSCs for up to 4 weeks was confirmed by MRI imaging and Prussian blue staining in the corpus cavernosa. CONCLUSIONS: Transplanted SPION-MSCs existed for up to 4 weeks in the cavernous nerve injured cavernosa of rats. Erectile dysfunction recovered and could be monitored by MRI.
Animals ; Contrast Media/pharmacology ; Dextrans/*pharmacology ; Disease Models, Animal ; Drug Delivery Systems/methods ; *Erectile Dysfunction/diagnosis/etiology/therapy ; Magnetic Resonance Imaging/methods ; *Magnetite Nanoparticles ; Male ; Mesenchymal Stem Cell Transplantation/*methods ; Monitoring, Physiologic/methods ; Penis/*innervation ; *Peripheral Nerve Injuries/complications/diagnosis/physiopathology/therapy ; Rats ; Suspensions ; Treatment Outcome

OBJECTIVETo evaluate the effect of elective microscopic resection of dorsal penile nerves in the treatment of primary premature ejaculation (PPE).

METHODSSeventy-eight PPE patients received elective microscopic resection of dorsal penile nerves, 5 branches in 9 cases, 6 in 17, 7 in 15, 8 in 14, 9 in 8, 10 in 6, 11 in 6, and 12 in 3. The patients were followed up for 12 months, and their intravaginal ejaculation latency time (IELT) and sexual intercourse satisfaction scores were recorded before and after treatment.

RESULTSCompared with the baseline, the IELT was significantly prolonged after surgery ([0.86 +/- 0.32] vs [6.65 +/- 3.9] min, P < 0.01), and the sexual intercourse satisfaction scores of the patients were dramatically increased (7.32 +/- 2.52 vs 12.32 +/- 3.76, P < 0.01), so were those of their sexual partners (4.46 +/- 1.36 vs 12.73 +/- 1.45, P < 0.01).

CONCLUSIONElective microscopic resection of dorsal penile nerves is safe and effective for the treatment of PPE.

Coitus ; Humans ; Male ; Patient Satisfaction ; Penis ; innervation ; Premature Ejaculation ; surgery ; Pudendal Nerve ; surgery

Recently, with application of evoked potentials technology in the test of somatic and autonomic nerves, quantitative sensory testing in the detection of small nerve fiber function, and functional magnetic resonance imaging in the detection of senior central function, the detection of neural function has become more accurate. This article reviews the progress and application of diagnostic methods about neurogenic erectile dysfunction in order to provide a reference for forensic diagnosis and research in the future.
Autonomic Nervous System/physiopathology* ; Autonomic Pathways/physiopathology* ; Erectile Dysfunction/physiopathology* ; Evoked Potentials/physiology* ; Humans ; Male ; Nervous System Diseases/complications* ; Neural Conduction ; Neurologic Examination/methods* ; Penile Erection/physiology* ; Penis/innervation* ; Sensory Thresholds

The injury of the penile cavernous nerve is a common cause of erectile dysfunction (ED). Reconstruction of the resected cavernous nerve can restore penile erectile function to normal. The methods for cavernous nerve repair include direct anastomosis, autotransplantation of the nerve, and substitution of the biodegradable artificial nerve, among which only autotransplantation of the sural nerve is used clinically at present. Besides, the nerve growth factor plays an important role in nerve reconstruction. This paper summarizes the methods of cavernous nerve reconstruction in the recent years.
Humans ; Male ; Nerve Regeneration ; Nerve Tissue ; surgery ; transplantation ; Neurosurgical Procedures ; methods ; Penile Erection ; Penis ; innervation

OBJECTIVETo establish a rat model mimicking erectile dysfunction following radical prostatectomy by crush injury or reaction of the cavernous nerve (CN).

METHODSThirty rats were randomized into CN crush group, CN resection group and sham-operated group. Four weeks after surgery, the rat models were evaluated by apomorphine test and ICP/MAP measurement. Fluorogold (FG) retrograde tracer was used to assess the CN injury. The penile tissues were then harvested for immunohistochemical detection of the nNOS-positive fibers to evaluate the CN injury.

RESULTSThe rats in CN crush group and CN resection group exhibited erectile dysfunction in apomorphine test or in response to electrical stimulation of the ganglion stellatum (MPG). In the sham-operated group, the rats showed normal erectile function with increased ICP/MAP following electrical stimulation (P<0.05). Immunohistochemistry revealed reduced nNOS-positive fibers in both CN crush group and CN resection group as compared with those in the sham-operated group (P<0.05), showing no significant difference between the former two groups (P>0.05). The FG-positive MPG cells in CN crush group and CN resection group were significantly less than that in the sham-operated group (P<0.05), and the positive cells were even less in CN resection group (P<0.05).

CONCLUSIONThe rat CN is structurally similar to human CN, and crush injury and resection of the CN are both reliable methods for establishing rat models of erectile dysfunction following radical prostatectomy.

Animals ; Disease Models, Animal ; Erectile Dysfunction ; etiology ; Male ; Nerve Crush ; Penis ; innervation ; Postoperative Period ; Prostatectomy ; adverse effects ; Rats ; Rats, Sprague-Dawley