Objective To establish the artificial bladder reflex arc by the normal body reflex pathway above the horizon of spinal cord injury to reinnervate the flaccid bladder and restore bladder micturition function. Methods An intradural microanastomosis was performed on the L6 ventral root tothe S2 ventral root. After axonal regeneration,the “patellar ligament-spinal cord center-bladder” reflex pathway was reestablished. A longterm function of the reflex arc was observed in the nerve electrophysiological experiment, detrusor electromyography experiment, and urodynamic testing 8 months after anastomosis. Results Trains of the stimuli(200 μV,5 ms) in the left L6 dorsal root and the nerve at the anastomosizedsite resulted in motor evoked potential from the disal to the anastomosized site before and after the spinal cord was destroyed horizontally between S1 and S4 segment levels in 2 Beegle dogs.The figure and amplitude of the evoked potential were similar to those of the control and general stability which showed anoninterventional wave. The urodynamic test revealed a rapid increase of the bladder pressure and a minor increase in the abdominal pressure. This showed that the bladder detrusor mainly resulted in the pressure increase.The bladder pressure increased to 60% of the normal on average compared with the controls when resulted in the left L6 dorsal root and the nerve anastomosized site were stinulated. Conclusion The long-term observation by the nerveelectrophysiological experiment, detrusor electromyography experiment, and urodynamic test indicate that the new artificial reflex arc can be established successfully. The somatic motor axons can regenerate into the parasympathetic endoneurial tubes of the autonomic nerve.
Bladder cancer is a common malignant tumor of the urinary system. The incidence and mortality of bladder cancer in China have been at a high level. In the past, people generally believed that the bladder was a sterile environment, but it has been found that there are symbiotic microorganisms in the bladder. In addition, microorganisms and their metabolites in urine may be involved in the occurrence and development of various urinary system diseases such as bladder cancer. At the same time, microorganisms and their component products such as Bacillus Calmette-Guerin vaccine play an important role in the treatment of bladder cancer. This article reviews the research progress of urinary tract microorganisms and the occurrence, development and treatment of bladder cancer, and aims to provide new ideas for the early diagnosis and treatment, prevention and prognosis evaluation of bladder cancer.
Objective To investigate the safety, efficacy and morbidity of onestage urethroplasty by using bladder mucosa for treatment of hypospadias. Methods From August 1991 to August 2003, 38 cases of congenital hypospadias were given bladder mucosa flap procedure and one stage urethroplasty. Results Thirty-eight cases of hypospadias treated with one stageurethroplasty by using bladder mucosa were followed up 6 months-9 years afterthe procedure. The success rate of the operation was 95%. Three cases of urethral fistula after the procedure were surgically repaired again, 2 cases of urethral stricture recovered after distension. The complication markedly lessened, micturation became normal with the reconstructed meatussituated at the proper site on the glands. Conclusion one stage urethroplastyby using bladder mucosa for treatment of hypospadias is a simple, effective andsafe surgery.
ObjectiveTo prepare a composite scaffold using bladder acellular matrix (BACM) and polyurethane (PU) for bladder repair and regeneration, and to evaluate its mechanical properties and biocompatibility. MethodsFresh bladder tissues were obtained from New Zealand rabbits and then treated with 1%SDS and 1%Triton X-100 to obtain BACM. The BACM was combined with PU to fabricate PU-BACM composite scaffold. The tensile strength and elongation at break of BACM and PU-BACM scaffolds were tested. Scaffolds and extracts of scaffolds were prepared to evaluate the biocompatibility. For cell-proliferation analysis, cell counting kit 8 method was used at 1, 3, 5, and 7 days after co-culture of human bladder smooth muscle cell (HBSMC) and scaffolds. The cell cycle was tested by flow cytometry after HBSMC co-cultured with extracts of scaffolds and DMEM culture medium (control group) for 24 hours. Finally, 12 New Zealand rabbits were used to establish the model of bladder repair and regeneration. Incision of 5 mm was made on the bladder, and PU-BACM scaffold was sutured with the incision. The rabbits were sacrificed at 10, 20, 40, and 60 days after surgery to observe the inflammatory cell infiltration, new tissues formation, and regeneration of epithelium by HE staining. ResultsThe tensile strength of BACM and PU-BACM composite scaffold was (5.78 ± 0.85) N and (11.88 ± 3.21) N, and elongation at break was 14.46%±3.21% and 23.14%±1.32% respectively, all showing significant diffeence (t=3.182, P=0.034；t=4.332, P=0.012). The cell-proliferation rates of controls, PU, BACM, and PU-BACM were 36.78%±1.21%, 30.49%±0.89%, 18.92%±0.84%, and 22.42%±1.55%, it was significantly higher in PU-BACM than BACM (P<0.05). In the bladder repair and regeneration experiment, inflammatory cell infiltration was observed at 10 days after operation, and reduced at 20 days after implantation. In the meanwhile, the degradation of scaffolds was observed in vivo. The regeneration of epithelium could be observed after 40 days of implantation. At 60 days after implantation, in situ bladder tissue formed. ConclusionPU-BACM composite scaffold has higher mechanical properties and better biocompatibility than BACM scaffold. PU-BACM composite scaffold will not lead to strong immune response, and new bladder tissue can form in the in vivo rabbit bladder repair experiment. These results can provide research basis and theoretical data for further study.
Previous studies have shown that growth arrest, dedifferentiation, and loss of original function occur in cells after multiple generations of culture, which are attributed to the lack of stress stimulation. To investigate the effects of multi-modal biomimetic stress (MMBS) on the biological function of human bladder smooth muscle cells (HBSMCs), a MMBS culture system was established to simulate the stress environment suffered by the bladder, and HBSMCs were loaded with different biomimetic stress for 24 h. Then, cell growth, proliferation and functional differentiation were detected. The results showed that MMBS promoted the growth and proliferation of HBSMCs, and 80 cm H2O pressure with 4% stretch stress were the most effective in promoting the growth and proliferation of HBSMCs and the expression level of α-smooth muscle actin and smooth muscle protein 22-α. These results suggest that the MMBS culture system will be beneficial in regulating the growth and functional differentiation of HBSMCs in the construction of tissue engineered bladder.
Objective To study major influential factors of the micturition alert device dedicated to neurogenic bladders for the product design and cl inical appl ication of the device. Methods One ferrite permanent magnet with thickness and diameter of 3 mm and 10 mm, respectively, and three NdFeB permanent magnets with the thickness of 3 mm and diameter of 10, 15 and 20 mm, respectively, were used. The effects of thickness of the abdominal wall as well as the position and type of permanent magnets on the micturition alert device dedicated to neurogenic bladders were measured in vitro simulated test, when the abdominal wall was set to 2, 3, 4, 5, 6, 7, 8 and 9 cm, respectively, and the position of permanent magnets was 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12 cm, respectively. The effect of the geomagnetic field on the device was measured under the condition that the thickness of the simulated abdominal wall was set to 2, 3, 4 and 5 cm, respectively,and the position of permanent magnets was 2, 3, 4, 5, 6, 7, 8, 9 and 10 cm, respectively. Results The value showed inthe warning unit was positively correlated with the position of the ferrite permanent magnet only when the thickness ofthe simulated abdominal wall was 2 cm (r=0.632, P lt; 0.05). The correlation between the value of the warning unit andthe position of NdFeB permanent magnets was significant (r gt; 0.622, P lt; 0.05), which was intensified with the increasingdiameter of NdFeB permanent magnets, but weakened with the increasing thickness of the simulated abdominal wall. The effect of the geomagnetic field was correlated with the exposition of the body, the position of the permanent magnet and the thickness of the abdominal wall. Conclusion The major influential factors of the micturition alert device dedicated to neurogenic bladder include the magnetism and location of the permanent magnet, the thickness of the abdominal wall and the geomagnetic field. These factors are correlated with and affect each other. Reasonable allocation of these factors may optimize the device.
Objective To observe whether umbilical cord mesenchymal stem cells (UCMSCs) can differentiate into the smooth muscle cells (SMCs) induced by bladder SMCs (BSMCs) conditioned medium so as to seek an alternative seed cells for the repair and reconstruction of the urology system. Methods UCMSCs and BSMCs were harvested from umbilical cord of full-term births and bladder tissues which were obtained from patients who underwent a radical cystectomy. BSMCs conditioned medium was prepared by mixing supernatant of BSMCs at passages 1-5 with complete medium at ratio of 1 ∶ 1. UCMSCs at passage 3 were cultured with BSMCs conditioned medium (induced group, group A) and complete medium (control group, group B), respectively; simple BSMCs served as positive control group (group C). The morphological changes of co-cultured UCMSCs were observed by inverted phase microscope, the expressions of α-smooth muscle actin (α-SMA), Calponin, and smooth muscle myosin heavy chain (SM-MHC) of UCMSCs were tested by immunofluorescence staining and Western blot at 7 and 14 days. Results The morphology of UCMSCs in group A started to change from a polygonal and short spindle shape to a large and spindle shape after co-culture, which was similar to BSMCs morphology; but the morphology of UCMSCs did not change obviously in group B. Immunofluorescence staining showed that the expressions of α-SMA, Calponin, and SM-MHC were positive in group C. At 7 days, the expression of α-SMA could be observed in groups A and B; at 14 days, the positive expression of α-SMA increased gradually in group A, but it did not increase in group B. At 7 days, a positive expression of Calponin could be observed in group A, and positive expression increased obviously at 14 days; the expression of Calponin could not be observed at 7 and 14 days in group B. However, the expression of SM-MHC could not be observed in groups A and B. The results of Western blot showed the expressions of α-SMA, Calponin, and SM-MHC protein were consistent with the results of immunofluorescence staining. Conclusion UCMSCs have the potential of differentiation into SMCs and may be a potential seed cells for bladder tissue engineering.
Objective To evaluate the feasibility of reconstructionof urothelium tissue in vivo using tissue-engineering technique. Methods The urothelium cells were obtained from young rabbit, bladder by mechanical and enzyme digested methods. After expanded in vitro, the 4th to 5th generation urothelium cells were seeded onto the surface of 8 Polylatical／glycolic acid copolymer polymer，the polymer matrix without seeding cells served as control group. A total of 8 cell-polymer scaffolds and 4 simply scaffolds were separately implanted into subcutaneous pockets of athymic mice. Theexperiment groups included cell-polymer scaffolds 4 weeks and cell-polymer scaffolds 8 weeks. The control group included simply scaffold 4 weeks and simply scaffold 8 weeks.After 4 and 8 weeks, the specimens were obtained and examined by gross inspection, histologically and immunohistochemically. Results The results of HE and Masson staining showed that the polymer were covered by urothelium cells layers and cells layers increased markly in experimental group. Immuocytochemical studies revealed that the cells were stained positively for anti-cytokeratins (AE1/AE3) in experimental group. Fiber tissue deposition were found on the surface of polymers in control group by HE and Masson staining. Immunocytochemical staining of implants showed the negative result for cytokeratins in control group. Conclusion It is feasibility that reconstruction of urothelium tissue using tissue-engineering -technique,whichprovides basic understandings for further development of the bladder and ureteral tissue engineered research.
ObjectivesTo evaluate the effects of Q-syte separating film needleless closed transfusion connector in flushing chamber of three-cavity urethral catheter.MethodsTo retrospectively analyze the patients who underwent transurethral resection of bladder tumor for non muscle-invasive bladder cancer from January 2015 to July 2016 in Zhongnan Hospital of Wuhan University. After terminating the continuous bladder irrigation, the observed group used Q-syte separating film needleless closed transfusion connector to seal the flushing chamber of three-cavity urethral catheter, and control group used conditional approach to connect drainage bag. The degree of comfort and satisfaction of patients, urinary tract infection, time of stopping bladder irrigation and bladder perfusion time between two groups were assessed.ResultsA total of 88 patients were included involving 63 (72%) males and 25 (28%) females with a mean age of 60.2±4.7 years. There were no significant differences between two groups in age, gender, BMI, and complications (P>0.05). Compared to control group, case group had higher level of comfort degree (mild discomfort: 86.4% vs. 25.0%, P<0.001; moderate discomfort: 13.6% vs. 52.3%, P<0.001; severe discomfort: 0.0% vs. 22.7%, P=0.001), satisfaction degree (97.9±2.1 vs. 84.5±3.9, P<0.001), and lower rates of urinary tract infection (11.4% vs. 29.5%, P=0.034). In addition, the case group spent shorter time in terminating bladder irrigation (50.48±1.78 vs. 207.74±5.41, P<0.001) and bladder perfusion (141.47±3.25 vs. 205.35±5.17, P<0.001). All differences were statistical significance.ConclusionsApplication of Q-syte separating film needleless closed transfusion connector for sealing flushing chamber of three-cavity urethral catheter after continuous bladder irrigation could promote the degree of comfort and satisfaction of patients, and decrease the rate of urinary tract infection, as well as the working efficiency of health care professionals.
Objective To investigate the feasibil ity of replacing urinary epithel ial cells with oral mucosa cell to reconstruct tissue engineered urethra by being seeded on bladder acellular matrix graft (BAMG). Methods Eighteen male New Zealand rabbits, aged 10 weeks, weighing 0.3-0.5 kg, were used in this study. Oral mucosa cell of 12 rabbits were isolated and seeded onto a culture dish with a feeder layer of 3T3 and a culture dish without 3T3, respectively. The morphologic change and growth condition of oral mucosa cells were observed by inverted phase contrast microscope after 2 days of seeding. The quantity of oral mucosa cells was counted using cell counting meter; the cell growth curve was drawn and the immunofluorescence staining with broad-spectrum keratin antibody was carried out. The bladders taken from the rest 6 rabbits were decelluled to make BAMG and the tissue of 1 cm × 1 cm was randomly selected to observe the effect of acellularization. The second passage oral mucosa cells cultured with 3T3 were appl ied to steril ized BAMG to obtain a issueengineered mucosa. The tissue-engineered mucosa was assessed using HE staining and scanning electron microscope after being cultured for 1 week. Results Oral mucosa cells seeded onto a feeder layer of 3T3 could be passaged for 7 or 8 generations with homogeneous forms and full function. Oral mucosa cells cultured without 3T3 could only be subcultured for 2 generations before aging and had multiple shapes and different sizes. Oral mucosa cells cultured by the two methods both started logarithmic growth on the 8th day and reached the peak value on the 14th day, which was indicated by the cell growth curve. However, more cells could be obtained through oral mucosa cells cultured with 3T3 than those cultured without 3T3. Oral mucosa cells manifestated green colour fluorescence cultured with or without 3T3. After the cells were removed, the BAMG presented as a porous membrane. The HE staining showed that the effect of acellularization was good and there were no cells at BAMG. The second passage oral mucosa cells cultured with 3T3 were expanded and seeded onto steril ized BAMG to obtain a tissue-engineered mucosa. Good compatibil ity of the compound graft was assessed using HE staining and scanning electron microscope. HE staining and scanning electron microscope showed that oral mucosa cells had good biocompatibil ity with BAMG after the tissue engineered mucosa was cultured for 1 week. Conclusion Oral mucosa cells of rabbit can be cultured in vitro and attain magnitude quantities. Oral mucosa cell also have good biocompatibil ity with BAMG and the compound graft could be a new material for urethral reconstruction.