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Background: Clinically isolated Staphylococcus epidermidis KK3-75, Enterococcus faecalis SMU-14, and Escherichia coli TF6-27 were subjected to test for bladder lodgments., Methods: Experimental cystitis on mice was investigated pathologically by light, confocal laser, and electron microscope after intravesical injection of cell suspensions of the strains. Bacterial affinities with bladder mucosal proteins and with extracellular components were detected by Western blot analysis., Results: Pathological findings of experimental cystitis revealed prominent infiltration of neutrophils except for those that were challenged with Enterococcus faecalis SMU-14. Capsule-like structures were demonstrated for the other two strains. Each strain showed histological tropism within the tissue sections, which correlated with the capability to bind the respective extracellular matrix components tested. Type I collagen bound only to cellular extracts of Enterococcus faecalis SMU-14 and Escherichia coli TF6-27, whereas fibronectin and type IV collagen bound only to those of Staphylococcus epidermidis KK3-75 and Enterococcus faecalis SMU-14. Bladder mucosal proteins had a variety of ability to bind cell surface proteins of each strain. Bacterial cell surface binding of all except for two of the bladder mucosal proteins detected was inhibited by extracellular matrix components., Conclusion: These experimental results suggest that the bacterial affinity of the bladder restricted by strain specific cell surface properties may be important for explanation in the difference of occurrence and progression of urinary tract infections caused by each strain.

We examined the effects of dimethyl sulfoxide (DMSO), hyaluronidase and saponin on drug absorption through the bladder mucosa. Female Wistar rats underwent intravesical instillation of each agent prior to the administration of lanthanum solution or anticancer drug; 4'-O-tetrahydropyranyldoxorubicin (THP). The absorption of lanthanum was recognized as dense grain in bladder tissue under electron microscope. The concentration of THP in bladder tissue and plasma was measured by high performance liquid chromatography. The ultrastructural change caused by instillation of DMSO, hyaluronidase and saponin was also investigated by transmission and scanning electron microscopy. The electron microscopic study of the bladder treated with DMSO revealed the exfoliation of superficial cells, and the electron-dense grains of lanthanum were observed in submucosal layer. The concentration of THP was twice as high as that of untreated animals in both bladder tissue and plasma. The treatment with hyaluronidase caused no severe histological change of bladder mucosa, and lanthanum was observed only in the cytoplasma of superficial cells. The increased concentration of THP was slightly exhibited in plasma but not in bladder tissue. The treatment with saponin caused vacuolization and swelling of superficial cells, and many grains of lanthanum were observed between superficial cells and intermediate cells. The concentration of THP in bladder tissue was significantly higher than that of untreated animals, but in plasma no difference was revealed. These findings indicate that each three agent caused an unique effect on drug absorption of bladder mucosa and such characteristic of each agent must be considered for clinical application. Saponin is thought to be useful on intravesical chemotherapy because of increased concentration of anticancer drug (THP) in bladder tissue without that in plasma.

Early changes of sugar chain detected by binding with lectins on luminal cell membrane of rat bladder epithelium treated with N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) were immunohistochemically examined with a transmission electron microscopy using ferritin-labeling lectins. BBN was given to male Wistar rats at the concentration of 0.05 W/V% in drinking water, and the rats were sacrificed at 4, 8, 16, 20 weeks, respectively, after the beginning of the BBN treatment. The bladder was treated with Half Karnovsky's solution via intra aortic cannulation for 20 minutes, and then removed. A part of bladder was examined grossly and by a light microscopy. The other part was cut into 3 x 3 mm squares and fixed with 2.5% glutaraldehyde solution and washed with 0.1 M cacogyl buffer with 10% sucrose. The tissue was sectioned and then incubated with ferritin-labeling lectins; concanavalin A (Con A), ulex europeaus agglutinin (UEA-1), peanut agglutinin (PNA) and dolichos biflorus agglutinin (OBA), respectively. These specimens were examined under a transmission electron microscopy. In cell membrane of the normal bladder, binding of Con A was found occasionally and binding of PNA was found focally, but binding of UEA-1 and DBA occurred incompletely on the cell membrane. The plaque and ridge on cell surface of bladder epithelium found to be obscured at 8 weeks after the treatment with BBN, and microvilli were barely observed under an electron microscope, but they increased gradually and could be seen by the microscope at the time of 16 weeks after the BBN treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

We evaluated histological changes induced by intravesical instillation of Bacillus Calmette Guerin (BCG) and doxorubucin hydorochloride (Adriamycin, ADM) for transitional cell carcinoma of the experimental bladder tumor. We administered rats N-butyl-N (4-hydroxybutyl) nitrosamine (BBN) by mixing it in the drinking water, and performed intravesical instillation of BCG, ADM or physiological saline solution as control in the 16th and 17th weeks and sacrificed in the 18th week after BBN exposure, and observed histological changes by light microscopy and electron microscopy. In the BCG group, the enlargement of the intercellular spaces of superficial tumor cells was found, and electron microscopic findings revealed a decrease or disappearance of the junctional complex. In the ADM group, hydrolytic and vacuolated cytoplasm was revealed. But such change as that in the BCG group was not found in the ADM and control groups. These results indicate that BCG may enlarge the intercellular spaces by changing the junctional complex, and lead to desquamate the tumor cells from the superficial cell layer.

The in vivo anticancer effects of high energy shock waves (HESW), on bladder cancer induced by N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN), were studied in the rat using the Siemens lithotriptor (Lithostar). There was no significant difference in the anticancer effects in delaying tumor growth, measured as the whole bladder weight including the weight of the cancer, between the groups with and without exposure to HESW. However, light microscopic examination revealed extensive submucosal bleeding and exfoliation of mucosa, and electron microscopic examination revealed degeneration of pleomorphic microvilli, swelling of mitochondria, and destruction of mitochondrial cristae in the cancer cells in bladders exposed to HESW. Flow cytometric determination of DNA content in the cancer cells exposed to HESW indicated a selective diminution of cells in the G2 and M phases and an increase of cells in the G0 and G1 phases of the cell cycle. While the mechanism of HESW-induced anticancer effect could not be determined on the basis of this study, these changes in the morphology and the cell cycle of cancer cells induced by HESW exposure suggest some kind of biological effects following exposure to HESW. It is expected that HESW be an effective method for the treatment of human cancer in combination with chemotherapy and radiation.

A normal rat bladder transitional epithelial cell line and seven tumor cell lines cultured from tumor induced by 0.05% N-Butyl-N-4-hydroxybutyl-Nitrosamine (BBN) were established and a comparative study on the doubling time and Ultrastructure of the cultured cell lines was performed. 1. Seven cultured cell lines (TU-B1 1-7) were obtained from bladder tumors induced by the administration of BBN for 32 weeks. 2. The doubling time of the cultured tumor cell lines were 43, 24, 28, 24, 25, 31, and 45 hours, suspectively. 3. These cultured tumor cell line were inoculated subcutaneously into nude mice and transitional epithelial carcinomas differing in degree of differentiation were observed. 4. Horizontal and tangential sections of these cultured tumor cell lines were examined by transmission electron microscopy. The development of intracellular organella and the presence of microvilli on the surface of the cells were noted. Intracellular adhesion was poor but the morphological characteristics of bladder transitional epithelium with compressed vesicles and other intracellular structures such as desmosome and free ribosomes were observed. 5. By applying the attached collagen gel method to our cell lines, it was possible to obtain cultured rat epithelial cells with the same morphology as in vivo, in addition to the tumor cell lines. 6. These cultured cell lines of normal rat transitional epithelium appear to be useful a experimental models with which one could clarify the function of the transitional epithelium and the mechanisms of bladder carcinogenesis.