Synthesis, characterisation and biological activity investigation of fullerenol/doxorubicin nanocomposite

Abstract

<p>U radu je predstavljena sinteza i karakterizacija&nbsp; novog fulerenol/doksorubicin nanokompozita, sintetisanog sa ciljem dobijanja potencijalne nove nanoformulacije postojećeg antineoplastika doksorubicina, koja bi pokazala veću biolo&scaron;ku aktivnost uz smanjenje neželjenih sporednih efekata koje sam lek izaziva, na prvom mestu kardiotoksičnosti.</p><p>Nanokompozit fulerenol/doksorbicin je okarakterisan brojnim&nbsp; metodama prateći dva osnovna eksperimentalna pristupa: molekulsko-spektroskopske metode (XPS, denzitometrija i transportne osobine, NMR, UPLC, Ramanska i UV-spektroskopija, SFM) i metode&nbsp; nanokarakterizacije (DLS, AFM, TEM), kao i računske simulacije(RDF). Osnovni cilj ispitivanja je bila detekcija postojanja&nbsp;nekovalentnog nanokompozita koji ostvaruju doksorubicin i fulerenolske nanočestice u vodenom rastvoru. Rezultati karakterizacije jasno i nedvosmisleno ukazuju na postojanje nekovalentnih interakcija unutar nanokompozita, &scaron;to dalje utiče na organizaciju i udruživanje čestica, a &scaron;to uslovljava i drugačiju biolo&scaron;ku aktivnost takvog sistema u odnosu na pojedinačne komponente.&nbsp; Rezultati biolo&scaron;kih ispitivanja na&nbsp;<em>in vitro</em>&nbsp;modelu različitih tumorskih ćelijskih linija pokazuju&nbsp; značajan&nbsp;antiproliferativni efekat nanokompozita, kao i selektivnost prema tumorskim u odnosu na zdravu ćelijsku liniju. Eksperimenti na <em>in vivo</em> modelu zebrica potvrđuju smanjenje toksičnosti nanokompozita u poređenju sa lekom, primarno kardiotoksičnosti. Računske simulacije, mikroskopski i spektroskopski podaci, kao i rezultati&nbsp; <em>in vitro</em> i <em>in vivo</em> studija ukazuju na to da nekovalentne interakcije između fulerenola i doksorubicina mogu biti ključni korak u stvaranju sinergističkog sistema za dostavu leka u biolo&scaron;ki sistem.</p><p>Multipotentnost fulerenola kao nanonosača lekova i nespecifičnost strukture doksorubicina kao leka, ukazuje na to da bi fulerenol mogao biti efikasan nanonosač i drugih antineoplastika, &scaron;to daje prostora za unapređenje antitumorskih osobina lekova posredstvom istovremene administracije leka.</p>

<p>The focus of this thesis was the synthesis and characterization of a novel fullerenol/doxorubicin nanocomposite, with the aim to obtain a potential nanoformulation of antineoplastic drug doxorubicin, which would&nbsp; express greater biological activity and lower level of adverse effects than the drug itself, in the first place cardiotoxicity.</p><p>Nanocomposite fullerenol/doxorubicin was characterized by means of numerous methods&nbsp; following two main experimental approaches: molecular-spectroscopic methods (XPS, densitometry and transport properties, NMR, UPLC, Raman and UVspectroscopy, SFM) and mehods of&nbsp; anocharacterisation&nbsp; (DLS, AFM,&nbsp; TEM), as well as computer simulations (RDF). The goal of characterization was detection of&nbsp; non-covalent interactions within nanocomposite that are established between fullerenol nanoparticles and doxorubicin in aqueous solution. The results clearly indicate the existence of non- covalent interactions within nanocomposite that affect the organization and assembling of the particles, which further exhibit different biological activity of such a system in comparison to components themselves. Results of biological activity on <em>in vitro </em>model of different tumor cell lines show significant antiproliferative effect of&nbsp; nanocomposite, as well as selectivity towards tumor cell lines. Experiments conducted on <em>in vivo</em> zebrafish model confirm the lowering ofthe adverse effects of the drug, especially cardiotoxicity, in case when nanocomposite was applied. Computer simulations, microscopic and&nbsp; spectroscopic results combined with encouraging <em>in vitro </em>and <em>in vivo</em> results point out&nbsp; that non-covalent interactions between fullerenol nanoparticles and doxorubicin may present the keyrole in formation of a synergistic system for nanodrug delivery into biological system. Multipotential of fullerenol nanoparticles as a nanocarrier and non-specific structure of doxorubicin as a drug imply that fullerenol may serve as a efficient nanocarrier of numerous other antineoplastics, which further allows the improvement of antitumor properties of drugs withsimultaneous drug administration.</p>