Synthesis and application of gold nanoparticiples to MDA-MB-231 breast cancer cells

Cilj ovog diplomskog rada bio je utvrditi radiosensitizacijski efekt nano čestica zlata (NZ) na stanicama raka dojke MDA-MB-231. Sintetizirane su tri vrste NZ obavijene citratima, glutationom i amino-dekstranom. Njihova veličina određena je skenirajućim elektronskim mikroskopom i UV-Vis-spektrofotometrom i iznosila je oko 15 nm. NZ su bile aplicirane na MDA-MB-231 stanice i pratilo se kvalitativno i kvantitativno nakupljanje NZ u stanicama. Transmisijski elektronski mikroskop je pokazao nakupljanje NZ u vezikulama, a kvantitativnom analizom je utvrđena točna koncentracija NZ u stanicama. Najveća koncentracija NZ u stanicama je postignuta s NZ obavijenim amino-dekstranom. Stanice tretirane NZ bile su izložene γ-zračenju, te se proučavao radiosensitizacijski efekt NZ na MDA-MB-231 stanice. MTT-testom utvrđeno je smanjenje vijabilnosti stanica uslijed aplikacije NZ i γ-zračenja. Klonogenski test pokazao se kao jako osjetljivi test za utvrđivanje utjecaja γ-zračenja + NZ na citotoksičnost stanica. Međutim, zbog velike citotoksičnosti samog γ-zračenja radiosensitizacijski efekt nije mogao biti utvrđen.The aim of this work was to study the radiosensitization effect of three types of gold nanoparticles (GNPs) on MDA-MB-231 cancer cells in vitro. GNPs were synthesized in order to be coated with citrate, glutathione and amino-dextrane. Their size was determined using electron microscopy and UV-VIS spectrophotometer and was around 15 nm. GNPs were applied to a MDA-MB-231 cancer cell line and their accumulation in cells was investigated. Transmission electron microscopy demonstrated accumulation of GNPs in cytoplasmic vesicles, while the Mass Spectroscopy was used to demonstrate the highest uptake of amino- dextrane coated GNPs. Cells treated with GNPs were γ-irradiated and their radiosensitization effect was studied. Short-term MTT viability assay showed no cells sensitivity to the effect of ionizing radiation, whereas the long-term clonogenic assay showed high sensitivity to the cytotoxic effect of γ-irradiation. However, the high cytotoxicity of γ-irradiation alone to cancer cells did not allow the determination of the radiosensitization effect of synthesized GNPs