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Preverjali smo, ali so morfološke spremembe pri apoptozi in feroptozi, vidne z vrstičnim elektronskim mikroskopom, dovolj značilne in jasne, da lahko zgolj na podlagi le-teh opredelimo tip smrti opazovane celice. Prav tako želimo ugotoviti, ali superparamagnetni železovi oksidni nanodelci (SPIONi) povzročijo celično smrt, ki ima molekularne in morfološke lastnosti, ki definirajo feroptozo. Celice HUVEC smo izpostavljali staurosporinu, ki je induktor apoptoze, RSL3, ki je induktor feroptoze ter SPIONom. Izvedli smo test citotoksičnosti, merili pa smo tudi vpliv staurosporina, RSL3 in SPIONov na specifično aktivnost acetilholinesteraze (AChE), stopnjo lipidne peroksidacije ter stopnjo apoptoze. Morfološke spremembe površine izpostavljenih celic smo opisali z vrstično elektronsko mikroskopijo. AChE se ni izkazala kot uporaben biomarker za ovrednotenje celične smrti, stopnja lipidne peroksidacije kot biomarker feroptoze pa je bila povišana pri tretiranju s RSL3 in SPIONi. Test stopnje apoptoze se je izkazal kot premalo specifičen, saj ni omogočal razločevanja med različnimi vrstami celične smrti. Staurosporin je na površini celic povzročil morfološke spremembe značilne za apoptozo, na celicah, izpostavljenih RSL3 so bile opazne morfološke značilnosti, ki jih literatura ne opisuje kot značilne za feroptozo. Tudi celice, izpostavljene SPIONom, niso imele za apoptozo in feroptozo-značilnih morfoloških značilnosti. Predvidevali smo, da SPIONi aktivirajo kombinacijo različnih mehanizmov celičnih smrti. The purpose of this master thesis was to find out if the morphological changes due to apoptosis in ferroptosis that we can observe with scanning electron microscope (SEM) are significantly different from eachother, which would show that we can use SEM as a method to differentiate between apoptosis in ferroptosis. Furthermore, we wanted to know if SPIONs cause a type of cell death that is morphologically and molecularly similar to ferroptosis. HUVEC cells were treated with inductor of apoptosis, staurosporin and inductor of ferroptosis, RSL. We also treated cells with SPIONs so we could determine what type of cell death they induced. We tested cells for cytotoxicity, AChE specific activity, lipid peroxidation and apoptosis. Treated cells were observed with SEM to describe morphological characteristics of their surface. AChE has not proven as a biomarker for a specific type of cell death. Biomarker for ferroptosis, lipid peroxidation, was, on the contrary, elevated in cells, treated with RSL3 and SPIONs. Test for apoptosis was not specific enough for differentiation between different cell deaths. Staurosporin induced morphological characteristics were specific for apoptosis, cells, treated with RSL on the other hand, were exhibiting morphological characteristic previously not described in the literature. Cells, treated with SPIONs, also had a unique morphology, not typical for apoptosis or ferroptosis |
