Evaluation of cytotoxicity, apoptosis, and angiogenesis induced by Kombucha extract-loaded PLGA nanoparticles in human ovarian cancer cell line (A2780)

The aim of this study was to load kombucha-fermented extract (KFE) on PLGA nanoparticles (KFE-PNPs) to increase bioavailability and to evaluate its anti-cancer effects. The KFE-PNPs (water(1)/oil/water(2)) were characterized using scanning electron microscope (SEM), Fourier-transform infrared spectroscopy (FTIR), and dynamic light scattering (DLS) assays, followed was measured the encapsulation efficiency (%EE) and release of KFE by UV spectrophotometer. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) procedure was used for investigation of KFE-PNPs toxicity, and then, the pro-apoptotic capacity of KFE-PNPs was evaluated by acridine orange (AO) and propidium iodide (PI) staining, flow cytometry, and molecular analysis for P53 and TNF-alpha genes. The angiogenic effect of KFE-PNPs was evaluated using chick chorioallantoic membrane (CAM) and real-time PCR (VEGF gene expression) methods. The DLS results showed the formation of stable particles (zeta potential: - 26.27 mv) in nanometer dimensions (288.32 nm) with uniform dispersion index (PDI: 0.3). The %EE of KFE in PLGA-NPs was reported to be 71%. The selective toxicity effect of KFE-PNPs against A2780 (IC50 < 200 mu g/mL) in comparison with HFF (IC50 > 500 mu g/mL) cells was reported. The pro-apoptotic effects of KFE-PNPs were confirmed by increasing the number of apoptotic cells in the AO/PI staining, increasing the percentage of SubG1 phase cells in flow cytometry, and increasing the expression of apoptotic genes (P53 and TNF-alpha). Decreased expression of VEGF gene in qPCR procedure as well as decreased length and number of blood vessels and embryonic growth factors in CAM assay showed anti-angiogenic effects of KE-PNPs. According to the results, KFE-PNPs can be suggested for more research on cancer therapy.

Automated summary

The aim of this study was to load kombucha-fermented extract (KFE) on PLGA nanoparticles (KFE-PNPs) to increase bioavailability and to evaluate its anti-cancer effects. The results showed that KFE-PNPs had stable nanoparticle characteristics, selectively killed cancer cells, promoted apoptosis, and inhibited angiogenesis.