Urothelium removal does not impact mucosal activity in response to muscarinic or adrenergic receptor stimulation.

The inner lining of the urinary bladder (urothelium and lamina propria, or bladder mucosa) has an important role as a tissue barrier between stored urine and the underlying smooth muscle, as well as in the modulation and regulation of bladder contractility. However, the individual influence of the apical urothelial layer on the contractile activity of this tissue is uncertain. The aim of this experiment was to identify the contractile activity of the lamina propria after removal of the urothelium. Several methods were used to mechanically disrupt the urothelium, including dabbing the tissue with a paper towel, longitudinal swipes with a cotton bud, or a longitudinal scrape with the edge of a scalpel. Hematoxylin-eosin staining was utilized to determine the level of removal of the apical urothelial cells. Spontaneous contractile activity was measured in organ baths, and responses to the agonists carbachol and isoprenaline were obtained. Three longitudinal swipes with a cotton bud was found to be the optimal method to remove the majority of the urothelium without damaging the lamina propria. Upon removal of the urothelium, the spontaneous activity of the tissue was unaltered. Similarly, responses to carbachol (1 µM) and isoprenaline (1 µM) were not affected after removal of the urothelium. The urothelium can be effectively removed without damaging the lamina propria. This apical tissue layer is not responsible for mediating the increases to spontaneous phasic activity or tonic contractions of the bladder mucosa (urothelium with lamina propria) when muscarinic or adrenergic receptors are stimulated. This research presents the lamina propria as the important cell layer mediating the overall contractile activity of the bladder wall.

Ventral onlay graft urethroplasty using bladder mucosa in a cat with a urethral stricture.

A 3-year-old, 4.0 kg, intact male domestic shorthair cat presented with postoperative dysuria following urethral resection and anastomosis for urethral rupture. Retrograde urethrography revealed a stricture in the pelvic urethra. Urethroplasty with bladder mucosa was performed following a bilateral pubic-ischial osteotomy. The bladder wall was resected to harvest an appropriately sized bladder mucosa graft. The graft was placed over the urethral defect in patch fashion and stabilized with interrupted sutures. The cat was able to urinate normally with no evidence of lower urinary tract signs 2 year postoperatively. Therefore, urethroplasty with an onlay bladder mucosa graft may be a feasible alternative to prepubic or subpubic urethrostomy for the treatment of pelvic urethral stricture in cats.

[Molecular Diagnostics of the Involvement of Visually Normal Mucosa in the Malignancy Process in Urothelial Bladder Cancer].

Immunofluorescent method by flow cytometry was used to quantify the expression of the tumor-associated protein ßIII-tubulin (TUBB3) in the tissue of urothelial bladder cancer and visually normal mucosa (56 samples in total). The expression of the marker was detected in 100% of cases, and heterogeneity of the TUBB3 expression level both in tumor tissue and in "normal" mucosa was revealed. The level of TUBB3 in the "normal" mucosa did not depend on the distance from the tumor (1 cm or more than 3 cm) and, on average, it was lower than in the tumor tissue (21.8 ± 10.8% and 24.9 ± 13.2% vs 35.2 ± 12.4%; p = 0.04 and 0.005, respectively). An increase of the TUBB3 expression in the tumor and in the "normal" mucosa was revealed in muscle invasive bladder cancer compared to non-muscle invasive bladder cancer. Therefore, in urothelial bladder cancer, the tumor-associated protein TUBB3 is a molecular marker of bladder mucosa involvement in the malignancy process and predicts the risk of tumor muscle invasion, which may influence indications for early cystectomy.

The Dependence of Urinary Bladder Responses on Extracellular Calcium Varies Between Muscarinic, Histamine, 5-HT (Serotonin), Neurokinin, Prostaglandin, and Angiotensin Receptor Activation.

With many common bladder diseases arising due to abnormal contractions, a greater understanding of the receptor systems involved may aid the development of future treatments. The aim of this study was to identify any difference in the involvement of extracellular calcium (Ca2+) across prominent contractile-mediating receptors within cells lining the bladder. Strips of porcine urothelium and lamina propria were isolated from the urinary bladder dome and mounted in isolated tissue baths containing Krebs-bicarbonate solution, perfused with carbogen gas at 37°C. Tissue contractions, as well as changes to the frequency and amplitude of spontaneous activity were recorded after the addition of muscarinic, histamine, 5-hydroxytryptamine, neurokinin-A, prostaglandin E2, and angiotensin II receptor agonists in the absence and presence of 1 µM nifedipine or nominally zero Ca2+ solution. The absence of extracellular Ca2+ influx after immersion into nominally zero Ca2+ solution, or the addition of nifedipine, significantly inhibited the contractile responses (p < 0.05 for all) after stimulation with carbachol (1 µM), histamine (100 µM), 5-hydroxytryptamine (100 µM), neurokinin-A (300 nM), prostaglandin E2 (10 µM), and angiotensin II (100 nM). On average, Ca2+ influx from extracellular sources was responsible for between 20-50% of receptor-mediated contractions. This suggests that although the specific requirement of Ca2+ on contractile responses varies depending on the receptor, extracellular Ca2+ plays a key role in mediating G protein-coupled receptor contractions of the urothelium and lamina propria.

Bladder mucosa-associated lymphoid tissue lymphoma progressed from chronic cystitis along with a comparative genetic analysis during long-term follow-up: a case report.

The pathogenesis of bladder marginal zone/mucosa-associated lymphoid tissue (MALT) lymphoma, which is the most common type of primary bladder lymphoma, has not been clarified. There are no reports that described histological and molecular time course of MALT lymphoma occurring in the bladder and the importance of the score on the Pelvic Pain and Urgency/Frequency (PUF) patient symptom scale during and after radiation therapy (RT). We present a case of MALT lymphoma with long-term comparative genetic analysis. A 77-year-old Japanese woman with hematuria and severe perineal pain was found to have a tumor-like lesion in the bladder trigone. She was diagnosed with cystitis based on the results of pathological examination and immunostaining after transurethral resection of the lesion. The second transurethral resection procedure was performed approximately 4 years after the first procedure because of recurrence of the hematuria and enlargement of a lesion in the left bladder wall. Postoperative pathologic examination confirmed a diagnosis of MALT lymphoma. Genetic analysis of immunoglobulin heavy chain (IGH) gene rearrangements showed more clonal progression from the first biopsy to the second. The patient then underwent RT, during which her perineal pain was exacerbated by radiation cystitis but finally decreased to a level less severe than that before treatment. The PUF patient symptom scale was useful to monitor her pain throughout the clinical course. No recurrence was detected more than 2 years after completion of RT.

Strategies to Get Drugs across Bladder Penetrating Barriers for Improving Bladder Cancer Therapy.

Bladder cancer is a significant public health concern and social burden due to its high recurrence risk. Intravesical drug instillation is the primary therapy for bladder cancer to prevent recurrence. However, the intravesical drug therapeutic effect is limited by bladder penetrating barriers. The inadequate intravesical treatment might cause the low drug concentration in lesions, resulting in a high recurrence/progression rate of bladder cancer. Many strategies to get drugs across bladder penetrating barriers have been developed to improve intravesical treatment, including physical and chemical methods. This review summarizes the classical and updated literature and presents insights into great therapeutic potential strategies to overcome bladder penetrating barriers for improving the intravesical treatment of bladder cancer.

[Electron microscopic characterization of the mucous membrane of the bladder with interstitial cystitis/painful bladder syndrome in the experiment].

OBJECTIVES: the study of the nature and severity of the reorganization of the structural elements of the bladder wall on experimental models of IC / BPS at the optical and ultrastructural levels. MATERIALS AND METHODS: The experimental model of IC / SBMP was created on 22 white New Zealand female rabbits weighing 1500-2000 g. The animals were divided into 2 groups: group 1 - 15 rabbits, which were introduced into the bladder wall urine taken from the animals bladder; Group 2 (control) - 7 animals that were injected into the bladder wall with a 0.9% NaCl solution. The biomaterial was examined by electron microscopy. The structural elements of the lamina propria of the bladder mucosa were evaluated. RESULTS: In the bladder mucosa of the experimental model, perivascular infiltration by inflammatory cells, pronounced edema of the lamina propria of the bladder mucosa, and the presence of numerous plasma cells having close contacts with macrophages, fibroblasts, and lymphocytes were revealed. In the nucleus of lymphocytes of a peripherally located supercondensed heterochromatin showed their apoptotic state. Edematous fluid was determined, penetrating mainly through the fenestra, located in the peripheral parts of the endothelial cells of capillaries and postcapillary venules. CONCLUSION: The study of the structural elements of the lamina propria of the bladder mucosa in the experiment by the method of electron microscopy made it possible to identify changes caused by the inflammatory process, both of an acute and productive nature. The results obtained showed that toxic damage to the lamina propria of the bladder mucosa causes apoptosis of fibroblasts of the lamina propria, leads to loosening of collagen fibers and, ultimately, to a decrease in the protective factors of the mucous membrane.

Villous Adenoma Arising in the Native Bladder Mucosa and the Upper Urinary Tract With Coexisting Neuroendocrine Carcinoma Following Augmentation Cystoplasty.

Villous adenomas arising in the bladder following augmentation cystoplasty procedures are exceedingly rare. Even rarer is their occurrence in the native bladder mucosa and the upper urinary tract. In this article, we present a unique case of multifocal recurrent villous adenoma involving native bladder mucosa of an augmented bladder, bilateral ureters, and renal pelvis, with coexistent foci of adenocarcinoma and neuroendocrine carcinoma, in a patient with history of augmentation colocystoplasty. We additionally discuss the pathogenesis of development of carcinoma in the setting of augmentation cystoplasty.

Aging increases the expression of vasopressin receptors in both the kidney and urinary bladder.

AIMS: The goal of this study was to determine whether aging effects the expression of V1a and V2 vasopressin receptors in the urinary bladder mucosa (UBM) and kidney. METHODS: UBM and kidneys were obtained from young (3 months-of-age) and old (25-30 months-of-age) female Fisher 344 rats. Tissue samples were analyzed by western blotting for V1a and V2 receptor expression, and rat plasma levels of vasopressin levels were measured by ELISA. RESULTS: V1a and V2 receptors were detected in both the UBM and kidneys. Aging significantly (P < 0.05) increased the expression of V2 receptors by 2.80 ± 0.52 and 6.52 ± 1.24-fold in the UBM and kidneys, respectively. Aging also increased V1a receptor expression in the kidneys (5.52 ± 1.05 fold; P < 0.05), but not in the UBM. To the best of our knowledge, because this is the first detection of V2 receptors in the mammalian bladder mucosa, we also probed human UBM for V2 receptors and observed high expression in human UBM. Unlike V1a and V2 receptors, aging had only a minor effect on plasma vasopressin levels (8% increase). CONCLUSIONS: V2 receptors are substantially increased in the aging UBM. The role of these receptors in UBM is as yet undefined, but given their presence and action in the kidneys, the possible effect of these receptors in free water regulation should be considered. The large age-related increase in the expression of V2 receptors in both the UBM and kidney may contribute to the effectiveness of desmopressin in age-related nocturia.

Free bladder mucosa graft harvested by water-jet: A novel, minimally invasive technique for urethral reconstruction.

The aim of the current study was to describe a novel approach of urethral reconstruction through minimally invasive harvesting of the bladder graft via endoscopic sub-mucosal dissection of water-jet. The records of two patients were reviewed, who underwent transurethral endoscopic surgical bladder mucosa graft harvest by water-jet and urethral reconstruction with informed consent. Case 1 was a 35-year-old male with anterior urethral stricture; case 2 was a 22-year-old male with secondary anterior urethral stricture and hypospadias following a failed hypospadias surgery. The two male patients successfully underwent urethral reconstruction using bladder mucosa graft harvested via endoscopic assisted by water-jet; no perforation, cysthemorrhagia or any other postoperative bladder-related complication was observed. Voiding cystourethrogram of case 1 indicated that the reconstructed urethra was unobstructed, and no recrudescence was observed within 4 months of follow-up. In case 2, dysuria had disappeared completely within 1 month of follow-up, and the urethra plate was successfully reconstructed by first-stage. To the best of our knowledge, this is the first report to demonstrate urethral reconstruction using a bladder mucosa graft harvested by transurethral endoscopic sub-mucosal dissection, assisted by water-jet. Transurethral endoscopic surgery may provide a minimally invasive approach instead of the traditional open surgery for harvesting bladder mucosa graft. Urethral reconstruction conducted with bladder mucosa graft harvested via endoscopic sub-mucosal dissection assisted by water-jet is a feasible and safe method, and the short-term follow-up results are encouraging.

Bladder Mucosal Graft Vaginoplasty: A Case Report.

BACKGROUND: Female vaginoplasty reconstruction, by choice, is usually performed with adjacent tissue. However in some clinical conditions such as high urogenital confluence sinus, cloacal malformation with extreme vaginal hypoplasia, local tissue may not be available. When vaginal replacement is performed in pediatric patients intestinal segments is preferred to non-operative procedures that require continuative dilations. However mucus production, malignant transformation risk and diversion colitis are important side effects. TECHNIQUE: We present a nouvel technique for vaginoplasty in a female child presenting with an isolated urogenital sinus malformation without virilization. The patient at 20 months underwent vaginoplasty using tubularized bladder mucosal graft. RESULTS: Surgical procedure was devoid of complications. Pubertal development occurred at age of 15. She underwent regular follow up until 18 years of age. At this age we performed clinical evaluation: absence of vaginal introitus stenosis and good cosmetic results were observed. Then she underwent vaginoscopy with multiple biopsies. Pathology examination of the bladder mucosal graft evidenced a normal structure of the mucosa, with a stratified squamous epithelium. DISCUSSION: Different techniques are taken into account for vaginal reconstruction according to the severity and to the type of malformation. We describe the use of bladder mucosal graft with favorable results after long term follow-up.

The antitumor effect of the novel cancer-specific adenovirus Ad-VEGFR on bladder cancer in vitro and in vivo.

Bladder cancer is one of the most common cancers. Approaches that block tumor angiogenesis are a new therapeutic strategy for locally advanced or metastatic BC. VEGF/VEGFR signaling has been obviously and negatively correlated with the progression and invasion of cancer. In this study, we constructed the recombinant adenovirus vAd-VEGFR-3 to investigate its antitumor effector in vitro/vivo. First, we used the recombinant adenovirus vAd-VEGFR-3 to infect bladder cancer cells and then collected the cell culture supernatant to treat human umbilical vein endothelial cells (HUVECs). The proliferation, migration and apoptosis of HUVECs were respectively detected by MTT, transwell and Annexin V-FITC/PI double staining. In addition, mouse bladder mucosa was injured by trypsin, and the orthotopic transplantation model of human bladder cancer was successfully constructed to clarify the anti-tumor effect of Ad-VEGFR in vivo. The results showed that Ad-VEGFR could inhibit the cancer's proliferation and migration, while promoting the apoptosis of HUVECs in vitro. Moreover, Ad-VEGFR could significantly promote the apoptosis of bladder cancer cells and then prevent tumor growth in vivo. In addition, it also down-regulated the expression levels of CD31, an endothelial cell marker which is closely related to the angiogenesis. Taken together, it suggests that the infection of adenovirus-carrying VEGFR in bladder cancer cells may inhibit blood vessel formation and prevent tumor progression.

From infection to immunotherapy:the immune response of bladder mucous membrane to host / 中国综合临床

Objective The pathogenesis of urinary tract infection and the preventive effect of Bacillus Calmette Guerin on bladder cancer imply the importance of bladder as a unique mucosal surface research. This article discussed that after bladder infected by bacteria,the way in which the microenvironment produce adaptive immunity. Once in the past, bladder is widely considered to be a sterile environment, the concept has been recently shaken. It is suggested that we should further study the influence of urinary tract pathogen infection or BCG immune response. In this paper, bladder biology has been illustrated, and bladder organ immunity is urgently needed to be explored and defined.

Urothelial cell expansion and differentiation are improved by exposure to hypoxia.

Cells obtained from a patient's biopsy have to be expanded after extraction to produce autologous tissues, but standard cell culture conditions often limit their growth or lifespan and could induce early and inadequate cell differentiation. Moreover, it has previously been reported that the air-liquid interface, that induces maturation of the urothelium, stimulated inadequate differentiation associated with aberrant keratin-14 expression. The aim of this study was to test the benefits of hypoxia during expansion of urothelial cells and maturation of the bladder epithelium in the context of tissue engineering. Bladder mucosa substitutes were reconstructed using the self-assembly method with urothelial cells (UCs) expanded in normoxia or hypoxia. Hypoxia improved UCs expansion until passage P7, whereas normoxic conditions limited the use of UCs to passage P4. Maturation of the urothelium was also compared in normoxic vs. hypoxic conditions. Using laminin V, p63, Ki-67, keratin-5 and -14, Claudin-4 and zonula occludens protein-1, we show a better organization of the basal UC layer in hypoxia despite a thinner intermediate layer. Finally, barrier function was assessed by permeation tests. Cell culture in hypoxia allowed the generation of bioengineered urological tissue closer to native bladder characteristics, which represents a promising avenue to circumvent the lack of adequate tissues for reconstructive surgery. Copyright © 2017 John Wiley & Sons, Ltd.

Ex vivo construction of a novel model of bioengineered bladder mucosa: A preliminary study.

OBJECTIVE: To generate and to evaluate ex vivo a novel model of bioengineered human bladder mucosa based on fibrin-agarose biomaterials. METHODS: We first established primary cultures of stromal and epithelial cells from small biopsies of the human bladder using enzymatic digestion and selective cell culture media. Then, a bioengineered substitute of the bladder lamina propria was generated using cultured stromal cells and fibrin-agarose scaffolds, and the epithelial cells were then subcultured on top to generate a complete bladder mucosa substitute. Evaluation of this substitute was carried out by cell viability and histological analyses, immunohistochemistry for key epithelial markers and transmission electron microscopy. RESULTS: The results show a well-configured stroma substitute with a single-layer epithelium on top. This substitute was equivalent to the control bladder mucosa. After 7 days of ex vivo development, the epithelial layer expressed pancytokeratin, and cytokeratins CK7, CK8 and CK13, as well as filaggrin and ZO-2, with negative expression of CK4 and uroplakin III. A reduction of the expression of CK8, filaggrin and ZO-2 was found at day 14 of development. An immature basement membrane was detected at the transition between the epithelium and the lamina propria, with the presence of epithelial hemidesmosomes, interdigitations and immature desmosomes. CONCLUSIONS: The present results suggest that this model of bioengineered human bladder mucosa shared structural and functional similarities with the native bladder mucosa, although the epithelial cells were not fully differentiated ex vivo. We hypothesize that this bladder mucosa substitute could have potential clinical usefulness after in vivo implantation.

[Inlaid labial versus bladder mucosal graft repair for complex urethral skin fistula].

OBJECTIVE: To compare the effect of inlaid labial mucosal graft repair (LMGR) with that of bladder mucosal graft repair (BMGR) in the treatment of complex urethral skin fistula after hypospadias repair. METHODS: This study included 55 cases of complex urethral skin fistula following hypospadias repair. We randomly assigned them to receive inlaid LMGR (n=36) or BMGR (n=19). After surgery, we compared the incidence of complications and recurrence rate of urinary fistula between the two groups of patients. RESULTS: The success rates of operation were 91.7% and 84.2% in the LMGR and BMGR groups, respectively, and the penile appearance was desirable in both groups. Postoperative complications included 2 cases of urinary fistula and 1 case of urethral stricture in each group. There were no statistically significant differences between the two groups in the success rate of operation (P>0.05) or the incidence rate of postoperative complications (P>0.05). CONCLUSIONS: Both inlaid LMGR and BMGR yield satisfactory results in the treatment of complex urethral skin fistula. However, LMGR involves less injury in mucosa collection and is easier to perform and therefore deserves more clinical attention.

Ventral bladder mucosa onlay graft urethroplasty for the treatment of panurethral stricture / 中华泌尿外科杂志

Objective To investigate the efficacy of ventral bladder mucosa onlay graft urethroplasty for the management of panurethral stricture.Methods From August of 2005 to July of 2013,11 cases of panurethral stricture were treated by ventral bladder mucosa onlay graft urethroplasty.The median age of the patients was 53 years (22-72 year),The median stricture length was 15 cm (12-18 cm).The patient was placed in the lithotomy position,Penile urethra was exposed by circumcoronal incision and degloving of skin,Bulbar urethra was exposed by inverted Y-shaped perineal incision.The strictured urethral segment was then opened ventrally in the midline up to at least 1 cm proximally into the healthy urethra.An appropriate size bladder mucosa graft was harvested,and was quilted to the splited urethra edge,the graft width was 1.5-2.0 cm.Two F10 fenestrated silicone catheters were left as urethral stents,a suprapupic cystostomy tube was left.The urethral stent was removed 4 weeks postoperatively.Follow-up was performed every 3 months for the first year,and annually thereafter.Success was defined as normal voiding with a maximum flow rates ≥ 15 ml/s,and the patients required no further instrumentation,including dilation or urethrotomy.Results The mean follow-up was 18 months (range,9-36 months),the overall success rate was 10/11.One patient developed urethral meatus stenosis 3 months postoperatively,and was managed by meatal dilatation.Conclusion Ventral bladder mucosa onlay graft urethroplasty can be used for the management of panurethral stricture,Bladder mucosa is an alternative substitution for complex urethral reconstruction.

Inlaid labial versus bladder mucosal graft repair for complex urethral skin fistula / 中华男科学杂志

<p><b>Objective</b>To compare the effect of inlaid labial mucosal graft repair (LMGR) with that of bladder mucosal graft repair (BMGR) in the treatment of complex urethral skin fistula after hypospadias repair.</p><p><b>METHODS</b>This study included 55 cases of complex urethral skin fistula following hypospadias repair. We randomly assigned them to receive inlaid LMGR (n=36) or BMGR (n=19). After surgery, we compared the incidence of complications and recurrence rate of urinary fistula between the two groups of patients.</p><p><b>RESULTS</b>The success rates of operation were 91.7% and 84.2% in the LMGR and BMGR groups, respectively, and the penile appearance was desirable in both groups. Postoperative complications included 2 cases of urinary fistula and 1 case of urethral stricture in each group. There were no statistically significant differences between the two groups in the success rate of operation (P>0.05) or the incidence rate of postoperative complications (P>0.05).</p><p><b>CONCLUSIONS</b>Both inlaid LMGR and BMGR yield satisfactory results in the treatment of complex urethral skin fistula. However, LMGR involves less injury in mucosa collection and is easier to perform and therefore deserves more clinical attention.</p>

Fournier gangrene with extensive necrosis of urethra and bladder mucosa: A rare occurrence in a patient with advanced prostate cancer.

Fournier gangrene (FG) is polymicrobial necrotizing infection of subcutaneous tissues and deep fascia, commonly involving the perineum, external genitalia, anterior abdominal wall and medial aspect of thighs. Urethral pathology, although often the inciting factor for FG, extensive involvement with urethral necrosis is very rare. This is the first report in English literature, describing complete sloughing of the bulbar urethra with ischemic necrosis of the bladder mucosa from FG. Such extensive disease is associated with high mortality, despite higher antibiotics, through debridement and intensive care. Urethral involvement needs extensive debridement and temporary or permanent urinary diversion.