GSK2126458 loaded polycaprolactone (PCL)/ polyvinylpyrrolidone (PVP) electrospun membranes: Production, characterization and anticancer activity

Skin cancer is one of the most dangerous cancers. The mutations resulting from the failure to repair DNA damage in skin cells result in the formation of cancer tissue in the skin. Skin cancer is a type of cancer that can spread to different parts of the human body and is divided into two main groups: melanoma and non-melanoma. The data in the literature indicate that the type of melanoma cancer accounts for 1% of cases of skin cancer. Although the incidence of melanoma skin cancer is low, patients have high mortality rates. There are different methods of treating skin cancer. The most common of these methods are biopsy and resection. In recent years, interest in the use of nanotechnology in the treatment of skin cancer has increased. Materials produced by nanotechnology are used as molecular carriers. Membranes produced using nanotechnological methods are intended to enable the drug to be applied directly to a specific area, to faster treatment and to reduce side effects. In this study, it was aimed to load GSK2126458 (Omipalisib®), which is used as a cancer drug, into Polycaprolactone (PCL)/ Polyvinylprolidone (PVP) fiber membranes obtained by electrospinning method and to investigate its effects on melanoma cancer cell line. Within the scope of the study, PCL/PVP/50 (50 µL drug and 50 µL DMSO), PCL/PVP/75 (75 µL drug and 25 µL DMSO), PCL/PVP/100 (100 µL drug) and PCL/PVP/C fiber membranes containing GSK2126458 drug and PCL/PVP/C fiber membranes without drug were produced by electrospinning method. DMSO ratio was kept constant in all samples. The morphology of the produced fiber structures were analyzed by scanning electron microscopy (SEM) and their chemical structures were analyzed by Fourier Transform Infrared Spectroscopy (FTIR). Fiber diameters and diameter distributions were calculated from the images obtained by SEM analysis using ImageJ software. Water retention properties of the fiber membranes were determined by water contact angle analysis and mechanical properties were determined by tensile analysis. Antibacterial properties of the fiber membranes were determined using E. coli, S. aureus, P. aeruginosa and MRSA bacterial strains. The IC50 value of the GSK2126458 on B16-F10 cell line was determined by resazurin assay. The effect of the drug on melanoma cells after being added to the fiber membrane was determined by viability analysis, SEM analysis of cell morphology and spreading behavior and immunofluorescent staining. As a result of the thesis study, GSK2126458 drug molecule loaded and drug-free control fiber membranes were obtained in a smooth and bead-free fiber structure. The average diameters of the fibers of PCL/PVP/C, PCL/PVP/50, PCL/PVP/75 and PCL/PVP/100 membranes were calculated as 0.6820,316, 0.5880.240, 0.6370.208 ve 0.6110.196 µm, respectively. Hydrophilic structures of the membranes were shown by water contact angles of 76.028.06, 83.631.75, 66.982.83 ve 82.9212.82 degrees for PCL/PVP/C, PCL/PVP/50, PCL/PVP/75 and PCL/PVP/100 fiber membranes, respectively. FTIR analysis showed specific peaks for PCL and PVP polymers, but a specific peak for the drug molecule could not be obtained because the drug was embedded in the polymer structure. The IC50 value of GSK2126458 on B16-F10 cell line was determined as 49.21 µM. When cell viability in drug-loaded membranes was analyzed compared to drug-free fiber membranes, it was determined that cell viability in PCL/PVP/50, PCL/PVP/75 and PCL/PVP/100 fiber membranes were 72.25%, 64.61% and 57.82% (p<0.05) for 24 hours, respectively. The viabilities were changed to 74.76%, 70.85% (p<0.05) and 60.19% (p<0.01) for 48 h, respectively. SEM images showed that cell spreading was more prominent in the control group than in the other groups. Immunofluorescence staining images obtained by staining fibrous actin protein of cells cultured on fiber membranes with anti F-actin antibody and nuclei with DAPI showed that cell density decreased with increasing amount of drug molecule. No significant changes were observed in the cell nucleus. In conclusion, it was shown that GSK2126458 drug loaded PCL/PVP fiber membranes were successfully produced within the scope of the thesis study and were effective on B16-F10 mouse melanoma cell line.