20.500.12556/RUL-186245 Vpliv nizkih ravni oksidativnega stresa na normalne urotelijske celice biomimetičnega in vitro modela intersticijskega cistitisa The effect of low levels of oxidative stress on normal urothelial cells in a biomimetic in vitro model of interstitial cystitis Intersticijski cistitis (IC) je kronično vnetje sečnega mehurja neznanega vzroka in z ne povsem jasno patofiziologijo. V raziskavi smo želeli vzpostaviti in podrobno analizirati nov biomimetični in vitro model IC. Uporabili smo kulturo normalnih urotelijskih celic prašiča (Sus scrofa domestica) in jih izpostavili nizkim ravnem oksidativnega stresa, ki smo ga sprožili z dodatkom encima glukaza oksidaze (GO) v hranilni medij za 24 ur. Raziskovali smo vpliv oksidativnega stresa na delno diferencirane (dd) in visoko diferencirane (vd) urotelijske celice. S testom viabilnosti smo kot najbolj primerno koncentracijo GO za posnemanje oksidativnega stresa določili 20 mE/mL. Z merjenjem transepitelijske upornosti (TER) in analizo permeabilnosti dekstrana-TRITC (70 kDa) smo proučevali vpliv oksidativnega stresa na prepustnost urotelija, z imunooznačevanjem pa njegov vpliv na izražanje dveh proteinov medceličnih stikov, okludina in E-kadherina. Z uporabo vrstične in presevne elektronske mikroskopije smo proučevali vpliv oksidativnega stresa na diferenciacijo in ultrastrukturo celic. Opazili smo statistično značilne razlike povečane prepustnosti dd celic po izpostavitvi oksidativnemu stresu, kar kaže na njihovo manjšo odpornost na oksidativni stres v primerjavi z vd celicami. Čeprav pri ostalih analizah nismo zaznali statistično značilnih razlik, so bili prisotni trendi, ki nakazujejo, da je oksidativni stres v vzpostavljenem urotelijskem modelu zmanjšal izražanje okludina in E-kadherina in znižal njegov TER. Prav tako smo opazili, da je oksidativni stres znižal stopnjo diferenciacije urotelijskih celic in vplival na nekatere njihove ultrastrukturne značilnosti. Interstitial cystitis (IC) is a chronic inflammatory disease of the urinary bladder with unknown etiology and unclear pathophysiology. In our study, we aimed to establish and analyse a new biomimetic in vitro model of IC. We used cultures of normal porcine urothelial cells (Sus scrofa domestica) and exposed them to low levels of oxidative stress for 24 hours. We studied the effects of oxidative stress on partially differentiated (pd) and highly differentiated (hd) urothelial cells. Using a cell viability assay, we determined the GO concentration of 20 mE/mL to be the most suitable for simulating the oxidative stress. The effects of oxidative stress on the permeability of the urothelium were determined by measuring transepithelial resistance (TER) and permeability analysis for dextran-TRITC (70 kDa), whereas the effects of oxidative stress on the expression of two intercellular junction proteins, occludin and E-cadherin, were assessed by immunolabeling. The effects of oxidative stress on cell differentiation and ultrastructure were analysed by scanning and transmission electron microscopy. We observed statistically significant increases in cell permeability of pd cells after exposure to oxidative stress, indicating that they are less resistant to oxidative stress than hd cells. Although other statistical analyses showed no significant differences, we observed trends, suggesting that oxidative stress reduces the expression of occludin and Ecadherin, lowers TER and negatively impacts differentiation and ultrastructural features of cells in the established urothelial model. biomimetični model intersticijski cistitis prašičje urotelijske celice oksidativni stres biomimetic model interstitial cystitis porcine urothelial cells oxidative stress true false false [M. Grm] Slovenski jezik Angleški jezik Magistrsko delo/naloga 2026-08-29 07:26:12 2026-08-29 07:26:20 2026-08-31 10:02:44 0000-00-00 00:00:00 2026 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 1970-01-01 616.62-002(043.2) 286186 289428227 60886.pdf 60886.pdf 1 6DE5B7C72C7D6EE2C27986B0E5106948 5e9f5aa75260fcc9f80b8c5af672db89c7468b689e47a0f7cd68e0f9dd746cf0 c038cacb-a369-11f1-9b0d-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=245383 Biotehniška fakulteta 0 0 0