Haocheng Lin

Nanoparticle Improved Stem Cell Therapy for Erectile Dysfunction in a Rat Model of Cavernous Nerve Injury

PURPOSE Recently intracavernous injection of stem cells has garnered great interest as a potential treatment of erectile dysfunction. However, most stem cells are washed out immediately after intracavernous injection. The goal of this study was to investigate using NanoShuttle™ magnetic nanoparticles to maintain stem cells in the corpus cavernosum after intracavernous injection, thereby improving stem cell therapy of erectile dysfunction in an animal model. MATERIALS AND METHODS Adipose derived stem cells were magnetized with NanoShuttle magnetic nanoparticles to create Nano-adipose derived stem cells. A total of 24 rats underwent bilateral cavernous nerve crush and were randomly assigned to 3 groups, including adipose derived stem cells, Nano-adipose derived stem cells and Nano-adipose derived stem cells plus magnet. Cells were tracked at days 1, 3, 5 and 9 after intracavernous injection. Another 40 rats with bilateral cavernous nerve crush were randomly assigned to 4 groups, including bilateral cavernous nerve crush, bilateral cavernous nerve crush plus adipose derived stem cell intracavernous injection, bilateral cavernous nerve crush plus Nano-adipose derived stem cell intracavernous injection and bilateral cavernous nerve crush plus Nano-adipose derived stem cell intracavernous injection plus magnet. Functional testing and histological analysis were performed 4 weeks after intracavernous injection. RESULTS In the in vitro study 1) NanoShuttle magnetic nanoparticles were successfully bound to adipose derived stem cells and 2) Nano-adipose derived stem cells migrated toward the magnet. In the in vivo study 1) cell tracking showed that Nano-adipose derived stem cells were successfully retained in the corpus cavernosum using the magnet for up to 3 days while most adipose derived stem cells were washed out in other groups by day 1 after intracavernous injection, and 2) intracavernous pressure/mean arterial pressure, and αSMA (α-smooth muscle actin) and PECAM-1 (platelet endothelial cell adhesion molecule 1) expression in the Nano-adipose derived stem cell group was significantly higher than in the other groups. CONCLUSIONS Magnetization of adipose derived stem cells with NanoShuttle magnetic nanoparticles kept adipose derived stem cells in the corpus cavernosum and improved adipose derived stem cell therapy of erectile dysfunction in an animal model.

New advances in erectile technology.

New discoveries and technological advances in medicine are rapid. The role of technology in the treatment of erectile dysfunction (ED) will be widened and more options will be available in the years to come. These erectile technologies include external penile support devices, penile vibrators, low intensity extracorporeal shockwave, tissue engineering, nanotechnology and endovascular technology. Even for matured treatment modalities for ED, such as vacuum erectile devices and penile implants, there is new scientific information and novel technology available to improve their usage and to stimulate new ideas. We anticipate that erectile technologies may revolutionize ED treatment and in the very near future ED may become a curable condition.

COX-2-10aa-PGIS Gene Therapy Improves Erectile Function in Rats after Cavernous Nerve Injury

INTRODUCTION Erectile dysfunction (ED) is a very common complication after radical prostatectomy. COX-2-10aa-PGIS is a newly engineered protein with COX-2 and prostacyclin synthase activities that converts arachidonic acid directly to prostacyclin (prostaglandin I2 [PGI2]). PGI2 is a potent smooth muscle relaxant. AIM The purpose of this study was to explore the effect and mechanism of COX-2-10aa-PGIS gene therapy in penile rehabilitation. METHODS Bilateral cavernous nerve crush (BCNC) in adult Sprague-Dawley rats was used to mimic radical prostatectomy-induced ED. Sprague-Dawley rats were randomly assigned into four groups: 1. sham surgery; 2. BCNC; 3. BCNC + null control recombinant adenovirus intracavernous injection; and 4. BCNC + Ad-COX2-10aa-PGIS intracavernous injection. Twenty-eight days later, intracavernosal pressure (ICP) was recorded under cavernous nerve stimulation; in the meantime, the mean arterial pressure (MAP) was monitored. At the end of the measurement, the penis was harvested and processed for (i) immunohistochemistry analysis of endothelial nitric oxide synthase (eNOS), alpha-smooth muscle actin (α-SMA), and transforming growth factor beta-1 (TGF-β1); (ii) Massons trichrome stain for smooth muscle/collagen ratios; (iii) Western blot of eNOS, α-SMA, TGF-β1, and COX2-10aa-PGIS; and (iv) terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay for apoptosis. MAIN OUTCOME MEASURES Erectile function was evaluated by ICP/MAP. Smooth muscle and endothelium functions in corpora cavernosum were assessed by Massons trichrome stain, immunohistochemistry, and Western blot. Apoptosis was identified by TUNEL assay. RESULTS The results were the following: 1. COX2-10aa-PGIS gene therapy improved erectile function (82%, compared with control) in the BCNC rat model; 2. COX2-10aa-PGIS gene therapy increased eNOS (121%) and α-SMA (118%) expression and decreased TGF-β1 (45%) expression; 3. COX2-10aa-PGIS gene therapy reduced cell apoptosis after cavernous nerve injury (64%); and 4. COX2-10aa-PGIS gene therapy improved smooth muscle/collagen ratios (81%). CONCLUSION Our data demonstrated that COX2-10aa-PGIS improved erectile function after cavernous nerve injury through antifibrotic and anti-apoptotic mechanisms.

Effect of androgen deprivation on the expression of aquaporins in rat prostate and seminal vesicles

The aim of this study was to investigate the level of secretions of prostate and seminal vesicles and its association with the expression of AQP0, 1, 4, 5, 6 and 8 in castrated rats. Eight‐week‐old male Sprague‐Dawley (SD) rats (n = 18) were randomly divided into control group, castrated rats group and castrated followed testosterone replacement group. Four weeks after surgery, the secretions and expression of AQP0, 1, 4, 5, 6 and 8 of prostate and seminal vesicles were determined. Serum testosterone was significantly lower in castrated groups than in control and testosterone replacement groups (P < 0.05). The level of prostate and seminal vesicle secretions and the expressions of AQP0, 1, 4, 5, 6 and 8 in prostate and seminal vesicles were significantly lower in castrated group than in control and castrated followed testosterone replacement groups (P < 0.05). The decreased prostatic and seminal vesicle secretions in castrated rats may be related to the decrease in AQP0, 1, 4, 5, 6 and 8 in prostatic tissue and seminal vesicles.

Testosterone replacement therapy and the risk of prostate cancer.

Understanding the role of testosterone replacement therapy (TRT) in the development and progression of prostate cancer is an important concept in treating patients with symptoms of hypogonadism. This article revealed a small number of mostly retrospective, observational studies describing the use of TRT in the general population, in men with prostatic intraepithelial neoplasia (PIN), in men with a history of treated prostate cancer, and in men on active surveillance for prostate cancer. The current literature does not report a statistically significant increase in the development or progression of prostate cancer in men receiving testosterone replacement for symptomatic hypogonadism, and the prostate saturation theory provides a model explaining the basis for these results. The use of TRT in men with a history of prostate cancer is considered experimental, but future results from randomized controlled trials could lead to a change in our current treatment approach.

Effects of estrogen deprivation on expression of aquaporins in rat vagina.

ObjectiveThis study aims to investigate the expression of aquaporin (AQP) 0, AQP3, AQP5, AQP6, AQP10, AQP11, and AQP12 in the vaginal tissue of ovariectomized rats. MethodsEight-week-old female Sprague-Dawley rats (n = 18) were randomly divided into three groups: control group (n = 6), ovariectomy group (n = 6), and ovariectomy/estrogen therapy group (n = 6). After 4 weeks, vaginal lubrication level and expression of AQP0, AQP3, AQP5, AQP6, AQP10, AQP11, and AQP12 in vaginal tissue were examined. ResultsSerum estrogen level was significantly lower in the ovariectomy group than in the control and ovariectomy/estrogen therapy groups (P < 0.05). Vaginal lubrication was significantly lower in the ovariectomy group (mean [SD], 1.62 [0.30]) than in the control group (mean [SD], 2.37 [0.70]) and ovariectomy/estrogen therapy group (mean [SD], 2.38 [0.73]; P < 0.05). Protein expression of AQP0, AQP3, AQP5, AQP6, AQP10, AQP11, and AQP12 was significantly lower in the ovariectomy group than in the control and ovariectomy/estrogen therapy groups (P < 0.05). ConclusionsDecreased vaginal lubrication in ovariectomized rats after electrostimulation may be partly caused by decreased AQPs in vaginal tissue.

Improving Erectile Function of Spontaneously Hypertensive Rats by Silencing ROCK2

OBJECTIVE To improve the erectile function of spontaneously hypertensive rats (SHRs) by silencing Rho-associated protein kinase 2 (ROCK2). METHODS Wistar-Kyoto rats (WKYs) and SHRs injected with 20-μL saline (WKY saline control and SHR saline control; n = 10) or 20 μL of 3 × 10(6) transducing units per milliliter negative control lentivirus (WKY negative control and SHR negative control; n = 10) were set as controls. After selecting the best inhibitory small interference ribonucleic acid (siRNA) by transducing 4 kinds of the lentiviral vector-based siRNA-targeting ROCK2 messenger ribonucleic acid (mRNA) respectively into cultured cavernous smooth muscle cells, 20 μL of 3 × 10(6) transducing units per milliliter of the lentiviral vectors were prepared and injected into the corpora cavernosa of WKYs (WKY siRNA; n = 10) and SHRs (SHR siRNA; n = 10). Seven days later, the maximum intracavernosal pressure to mean arterial pressure ratio (ICPmax/MAP), the expression levels of ROCK2, endothelial nitric oxide synthase (eNOS), and phosphorylated eNOS in the penis were measured and determined. RESULTS In cavernous smooth muscle cells of SHR culture, 3 kinds of ROCK2 siRNA significantly inhibited ROCK2 mRNA expression. The lentiviral vector-based siRNA-targeting ROCK2 mRNA at the 2287th nucleotide position significantly increased the ICPmax/MAP in the SHR siRNA group more than in SHR saline control and SHR negative control groups. There was no significant difference in the ICPmax/MAP among WKY saline control, WKY negative control and WKY siRNA groups. The ICPmax/MAP in the SHR siRNA group was significantly lower than that in the WKY saline control group. ROCK2 expression in the penis was significantly decreased in SHR siRNA group compared with that in SHR saline control and SHR negative control groups. The expression of eNOS and phosphorylated eNOS was significantly increased in SHR siRNA compared with that in SHR saline control and SHR negative control groups. CONCLUSION The gene therapy with lentiviral vector-based siRNA-targeting ROCK2 mRNA can significantly improve erectile function mainly by directly inhibiting ROCK2 pathway in the SHR.

Evaluation of the long-term safety and effectiveness of tadalafil once daily in Chinese men with erectile dysfunction: interim results of a multicenter, randomized, open-label trial

Once-daily tadalafil administration has been well established; however, studies about tadalafil once-daily treatment in the Chinese population are lacking. In this phase 4, postmarketing study, we ascertained the long-term safety and effectiveness of tadalafil 2.5 mg and 5.0 mg once daily in Chinese men with erectile dysfunction (n = 635). The primary endpoint of the study was safety at 12 months as assessed by the proportion of patients experiencing at least one treatment-emergent adverse event (serious or nonserious). The secondary endpoints included safety and effectiveness, measured by the International Index of Erectile Function-Erectile Function (IIEF-EF) domain scores. Similar adverse events to the known safety profile of tadalafil, such as nasopharyngitis, upper respiratory tract infection, headache, and dizziness, were detected. No new cardiovascular safety concerns were observed. After 3 months of treatment, significant increases in IIEF-EF domain scores were detected for both 2.5-mg (least squares [LS] mean change: 6.3; 95% confidence interval [CI]: 5.4–7.1; P < 0.001) and 5.0-mg (LS mean change: 7.4; 95% CI: 6.8–7.9; P < 0.001) tadalafil doses, and significance was maintained up to 12 months. In addition, approximately 40% of patients regained normal erectile function (IIEF-EF ≥26) following 1 year of tadalafil once-daily treatment. The findings in this study provide evidence for the extended effectiveness and tolerability of tadalafil, demonstrating no new safety concerns, in a Chinese population and make once-daily tadalafil administration a viable option for improving sexual performance and satisfaction in Chinese men with erectile dysfunction.

MP89-07 THE MECHANISMS OF NANOPARTICLE IMPROVING ADIPOSE DERIVED STEM CELLS THERAPY FOR ERECTILE DYSFUNCTON

determined diabetic rats randomly got intracavernous (IC) injection of phosphate buffer solution (PBS), ADSCs or MTs. Another eight normal rats equally received IC injection of PBS. MTs were generated with a hanging drop method and the injected cells were tracked in ADSCs and MTs injected rats. Four weeks after the treatments, intracavernous pressure (ICP), histopathological changes in corpus cavernosum (CC), and functional proteins were measured. Rat cytokine antibody array was used to detect ADSCs or MTs lysate. RESULTS: MTs expressed vascular endothelial growth factor (VEGF), nerve growth factor (NGF) and tumour necrosis factor-stimulated gene 6 (TSG-6). MTs injection had a higher retention than ADSCs injection and MTs treatment better improved ICP, neuronal nitric oxide synthase (nNOS) expression, smooth muscle and endothelial contents in diabetic rats, ameliorated local inflammation in CC. CONCLUSIONS: IC injection of MTs improves the erectile function and histopathological changes in streptozotocin-induced diabetic rats and appears to be more promising than traditional ADSCs. The underlying mechanisms involve increased cell retention accompanied with neuroprotection and anti-inflammatory behaviors of the paracrine factors.

Vacuum erectile device for penile rehabilitation

The vacuum erectile device (VED) uses negative pressure to increase blood inflow and oxygen into the corpora cavernosum, with a ring at the base of the penis to maintain the erection for intercourse or without a ring for penile rehabilitation. As the limitation of phosphodiesterase 5 inhibitors (PDE5I) showed in the treatment of refractory erectile dysfunction (ED), the use of VED resurged and is becoming the first-line therapy in treatment of ED after radical prostatectomy. Currently, the combination therapy of VED and PDE5I and of VED and intracavernous injection are advocated. Furthermore, there has been increasing interest in the use of VED to preserve penile length in inflatable penile prosthesis preoperation procedure and Peyronies disease. Hereby, we reviewed the underlying mechanisms, the status of VED in penile rehabilitation, the combination therapy and the expanded use of VED.