The aging society has a deep impact on patient care in urology. The number of patients in need of partial or whole bladder wall replacement is increasing simultaneously with the number of cancer incidents. Therefore, urological research requires a model of bladder wall replacement in adult and elderly people. Two types of porcine collagen I/III scaffolds were used in vitro for comparison of cell growth of two different pig breeds at different growth stages. Scaffolds were characterised with scanning electron and laser scanning microscopy. Urothelial and detrusor smooth muscle cells were isolated from 15 adult Göttingen minipigs and 15 juvenile German Landrace pigs. Growth behaviour was examined in cell culture and seeded onto the collagen scaffolds via immunohistochemistry, two-photon laser scanning microscopy and a viability assay. The collagen scaffolds showed different structured surfaces which are appropriate for seeding of the two different cell types. Moisturisation of the scaffolds resulted in a change of the structure. Cell growth of German Landrace urothelial cells and smooth muscle cells was significantly higher than cell growth of the Göttingen minipig cells. Seeding of scaffolds with both cell types from both pig races was possible which could be shown by immunohistochemistry and two-photon laser scanning microscopy. Growth behaviour on the scaffolds was significantly increased for the German Landrace compared to Göttingen minipig. Nevertheless, seeding with the adult Göttingen minipig cells resulted in a closed layer on the surface and urothelial cells and smooth muscle cells showed increasing growth until day 14. The results show that these collagen scaffolds are adequate for the seeding with vesical cells. Moreover, they seem appropriate for the use as an in vitro model for the adult or elderly as the cells of the adult Göttingen minipig too, show good growth behaviour.
One possible symptom of overactive bladder (OAB) is urinary incontinence, which is generally considered to be an age-associated disease and which is rapidly increasing with demographic changes. Rodent models are commonly used for the investigation of lower urinary tract functions, although the use of these species has limitations in several translational aspects. In biomedical research and preclinical toxicology, Göttingen minipigs are used increasingly. But in urological research, only few data are available for Göttingen minipigs. To the best of our knowledge, this study is one of the first to provide reference data of micturition in female Göttingen minipigs. Micturition frequency and volumes were monitored and analyzed in five female Göttingen minipigs. Voided volume was 520 ± 383 mL (mean ± standard deviation of mean [SD]) and micturition frequency 6.17 ± 3.68 (mean ± SD). We also performed a review of the literature to compare our data with data from different species (humans, pigs, rats and mice). Our findings revealed that micturition volume and frequency of Göttingen minipigs are more comparable with that of humans, leading to the conclusion that Göttingen minipigs may be the better choice for translational research in different research fields, such as urology, neurology and nephrology, etc. The provision of in vivo reference values meets with the 3R concept of 'reduction, refinement and replacement' of laboratory animals, because they allow comprehensive statistical power calculations (reduction), planning of telemetric approaches (refinement), and generation of computer-based modulation for the development of intravesical drug delivery systems (replacement).
European and German directives for approval of new medical devices require tests for cytotoxicity in relevant media, since urine can influence cytotoxicity of biodegradable devices. The aim of this study was to determine the long-term cytotoxicity of PLGA-b-mPEG (PLGA-PEG) polymer carriers and artificial urine (AU) to human UROtsa cells. Benign urothelial UROtsa cells were incubated in fetal bovine serum-containing RPMI 1640 medium supplemented with a range of concentrations of AU for 24 h and 7 days. Cell viability was determined by the XTT assay and by live/dead staining. The cytotoxicity of medium containing degradation products from PLGA-PEG carriers was also tested on the UROtsa cells in AU-containing and control medium. PLGA-PEG carriers exhibited no cytotoxicity to UROtsa cells after 24 h of incubation. However, after 7 days, cytotoxicity was observed, but this was largely attributable to the effects of 30% AU on the cells. Compared to phosphate buffer saline (PBS) and normalized to RPMI 1640 medium, significant cytotoxicity was observed by 24 h in medium containing 50% AU and by 7 days in medium containing 30% AU. Live/Dead staining confirmed proliferation results and no pH-changes could be observed. Here we demonstrate for the first time the impact of AU on standard cytotoxicity tests related to biomaterials for urinary-tract applications. Our study showed cytotoxic effects of high concentrations of 50% AU by 24 h and by physiological concentrations of AU (i.e., 30%) by 7 days. We have also demonstrated that PLGA-PEG has no cytotoxic effects in the appropriate AU-containing test environment. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2140-2147, 2018.
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