Hiperisin Yüklü Kitosan Nanopartiküllerin A549 Akciğer Kanser Hücrelerinde Etkilerinin Araştırılması

Abstract

Lung cancer is still continues to be one of the most common types of cancer today and is the leading cause of cancer death. Hypericin (HY) is a hydrophobic, natural anti-cancer component derived from the plant Hypericum Perforatum. HY is one of the preferred molecules in photodynamic therapy based on activation by light. HY causes cell death by the formation of oxygen-dependent reactive oxygen species (ROS) by photoreaction. HY has a relatively large molecular structure and shows clustering alone, as well as limited uptake and degradation in the physiological environment, which affects the response it will create. For these reasons, it is crucial to make HY water-soluble by using nanoparticles (NP), which are an important part of hydrophobic drug carrier systems. It is aimed that the anti-cancer effect expected to be seen with HY will provide a more specific and longer-term treatment approach by the virtue of NPs. Chitosan, which forms the polymeric structure of the nanoparticle, is a natural component obtained from chitin, but it stands out because of its biocompatibility, limited cytotoxicity, and easy solubility. It is seen as a potential anti-cancer tool by targeting tumor tissues with cancer drugs. Within the scope of the thesis, the anti-cancer property of A549, lung cancer epithelial cell, was evaluated, by direct HY and loaded with chitosan nanoparticles. The ionic gelation method was preferred for the synthesis of chitosan nanoparticles; its structure has been studied in scanning electron microscopy (SEM). HY concentrations of the determined values (200 nM, 400 nM, 600 nM) were added during synthesis and applied to cells. After 24 hours incubation, cells were exposed to light for 30 minutes at a dose of 6 lux/J cm2. Analysis was carried out after 24 hours at the end of the PDT period. Cell viability analysis was measured with MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) agent and the amount of reactive oxygen species formed was evaluated with DCFDA (2’,7’-Dichlorodihydrofluorescein diacetate) while cell death type was specified with AO/PI fluorescent staining. Cellular uptakes of NPs were imaged wih FITC (Fluorescein isothiocyanate) fluorescent dye and apoptosis formations with Tumortac kit have been studied. Among all experimental groups, NP groups were seen to increase the uptake of HY as targeted, with more increasing cell death. As a result of MTT analysis, up to 56% decrease in cell viability was observed in the HY-NP 600 nM group within 48 hours, while increasing the HY concentration in nanoparticle groups also increased the decrease in dose-related viability. When cell death type AO/PI was evaluated with fluorescent dyes, necrosis was observed in most cells. Characterization studies of the generated NPs were carried out by SEM analysis. By developing chitosan NPs, it has been suggested that the uptake of the active ingredient in cancer cells can be increased and side effects can be reduced by using herbal ingredients instead of drugs. It is promising for tumor tissues formed in lung cancer.