Salinomisin ve BnHAp Yüklü Doku İskelelerinin Antikanserojen ve Antibakteriyel Etkinliğinin Araştırılması
Özet
In the scope of the thesis, it is aimed to investigate the synergistic effect of an anticancer drug, salinomycin, loaded poly (lactic-co-glycolic acid) (PLGA 65:35) nanoparticles with the boron doped nanohydroxyapatite (BnHAp) that has antibacterial activity. For this purpose, following the synthesis of salinomycin loaded PLGA nanoparticles, characterization studies and in vitro release studies were carried out. Cell culture and bacterial culture studies were performed with salinomycin loaded PLGA nanoparticles in the presence of BnHAp.
PLGA nanoparticles with salinomycin content of 2.5% (w/v) were produced by emulsion preparation-solvent evaporation technique. The empty and salinomycin loaded PLGA nanoparticles (PLGA-SAL) produced have an average diameter of 185.8 nm and 187.4 nm, respectively. The encapsulation efficiencies of the PLGA-SAL nanoparticles were calculated to be 97.4 ± 1.87% according to the measurements done with the HPLC. In vitro release studies were carried out on PLGA-SAL nanoparticles in phosphate buffered saline (PBS) and the release of salinomycin reached 43% with a burst effect at the 1st hour. The sustained release of salinomycin from PLGA nanoparticles with controlled and regular increase reached 100% at the end of the 45th day. Subsequently, a release study was initiated with chitosan tissue scaffolds incorporating PLGA-SAL nanoparticles and BnHAp, and 44% of the salinomycin was released with a burst effect in the first 24 hours. The release lasted approximately 59 days. It was determined that the release of salinomycin from both particles and scaffold was consistent with the Hixson-Crowell release kinetic model.
In the second phase of the studies, cell culture studies with MG-63 osteosarcoma cells were performed. For this purpose, the control group (MG-63 cells), salinomycin, empty PLGA nanoparticles, PLGA-SAL nanoparticles, PLGA-SAL/BnHAp and PLGA-SAL/nHAp groups were included. During the cell culture studies, cell viability was examined by live-dead staining analysis, cytoskeletal organization of nanoparticles by Alexa Fluor/DAPI staining and intracellular uptake of nanoparticles by crystal violet staining. When PLGA-SAL nanoparticles were used at the optimal dose of 0.2 mg, apoptosis was seen in MG-63 osteosarcoma cells. However, cell viability was determined to be high in groups which are containing BnHAp and nHAp due to the proliferative effects of nHAP and BnHAp on the MG-63 cells.
Finally, bacterial culture studies were performed with methicillin resistant Staphylococcus aureus (MRSA-ATCC 43300), methicillin sensitive Staphylococcus aureus (MSSA-ATCC 29213) and control group Staphylococcus aureus (ATCC 25923) strains. In addition to salinomycin loaded antibiotic discs; the changes in inhibition areas in the presence of BnHAp were examined for 19 different antibiogram discs. In the culture studies using disc diffusion method (normal antibiogram), double disc synergy (punch) method and agar dilution method; the synergistic effect in the presence of BnHAp and the increase of inhibition area of salinomycin loaded antibiogram discs were observed.
