Physico-chemical characterization of fullerenol nanoparticles and biological analysis

Abstract

<p>U ovoj doktorskoj disertaciji su ispitivane fiziĉko-hemijske<br />karakteristike fulerenolskih nanoĉestica (FNP) i njihov potencijalni</p><p>radioprotektivni efekat na in vitro modelu K562 ćelija. Cilj prvog<br />dijela ovog istraţivanja je bio determinacija fiziĉko-hemijskih osobina<br />molekula fulerenola C₆₀(OH)₂₄, zatim karakterizacija fulerenola sa<br />aspekta nanoĉestice i u poslednjem segmentu determinacije fiziĉko-<br />hemijskih osobina je izvr&scaron;ena sinteza i karakterizacija kompleksa<br />FNP-a sa biolo&scaron;ki znaĉajnim metalima: mikroelementima gvoţĊem i<br />bakrom (Fe i Cu), i kalcijumom kao makroelementom (Ca). Fiziĉko-<br />hemijska karakterizacija je izvedena na velikom broju modela (u<br />smislu dijapazona koncentracija, vremenskih intervala, kombinacija<br />rastvaraĉa, pH vrijednosti, pona&scaron;anja u biolo&scaron;kim teĉnostima i<br />prisustva jona biogenih metala) kako bi se ispitao i prona&scaron;ao &scaron;to bolji<br />model i kombinacija navedenih faktora za ispoljavanje optimalne<br />biolo&scaron;ke aktivnosti. U drugom dijelu ovog rada su ispitivane biolo&scaron;ke<br />osobine fulerenolskih nanoĉestica u svojstvu citotoksiĉnog agensa i<br />radioprotektora na in vitro modelu ćelijske linije humane mijeloidne<br />eritroleukemije, pri jednokratnom ozraĉivanju visokom i niskom<br />dozom jonizujućeg zraĉenja. Putem odreĊivanja vijabilnosti,<br />sposobnosti formiranja kolonija i analizom morfologije tretiranih<br />ćelija, kao i odreĊivanjem aktivnosti antioksidativnih enzima je<br />definisan biolo&scaron;ki odgovor K562 ćelijske linije na primijenjenu visoku<br />dozu zraĉenja od 24 Gy u prisustvu fulerenolskih nanoĉestica. U cilju<br />odreĊivanja sposobnosti FNP da indukuje promjene ekspresije gena<br />koje dovode do modifikacije ćelijskog odgovora na radijacionu<br />povredu, ispitivana je ekspresija gena odgovornih za antioksidativni i<br />antiapoptotski odgovor kod ozraĉenih K562 ćelija i onih tretiranih<br />fulerenolskim nanoĉesticama pri primjeni malih doza jonizujućeg<br />zraĉenja od 2 Gy.</p>

<p>The physical and chemical characteristics of fullerenol nanoparticles (FNP) and their potential radioprotective effect on the in vitro model of K562 cells were examined in this doctoral dissertation. The aim of the first part of the study was determination of physico-chemical properties of fullerenol molecule C₆₀(OH)₂₄, followed by characterization of fullerenol in terms of nanoparticles, and, in the last part of physico-chemical characterization, the aim was to analyse properties of synthesized FNP&#39;s with biologically important metals: iron micronutrients and copper (Cu and Fe), and calcium as macroelement (Ca). Physico-chemical characterization was performed on a large number of models (in terms of range of concentration, time intervals, a combination of solvents, pH values, behaviors in biological fluids and the presence of biogenic metal ions) in order to investigate and found a better model and a combination of these factors for the optimal biological activity. In the second part of this research, examination aimed for the the biological properties of fullerenol nanoparticles as a radioprotector and cytotoxic agent in irradiated human myeloid erythroleukemia cell line, with high dose and low-dose of ionizing radiation. Biological response K562 cell lines to the applied high dose of 24 Gy radiation in the presence of fullerenol nanoparticles was defined by determination of cell viability, colony forming ability, the analysis of the morphology of the treated cells, and the activities of antioxidant enzymes. In order to determine the FNP ability to induce changes in gene expression that lead to the modification of cellular response to radiation injury, the expression of genes responsible for antioxidant and antiapoptotski response in irradiated K562 cells and those treated fulerenolskim nanoparticles in the application of low doses of ionizing radiation of 2 Gy, was analysed.</p>