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

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Abstract

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 (water1/oil/water2) 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-α 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 µg/mL) in comparison with HFF (IC50 > 500 µg/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-α). 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.

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Acknowledgements

The authors very much appreciate the support provided by the Islamic Azad University, Shahrood, Iran, in the conducting of the present research.

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Authors and Affiliations

  1. Department of Chemistry, Shahrood Branch, Islamic Azad University, Shahrood, Iran

    Sara Ghandehari & Jafar Izadi Nia

  2. Department of Biochemistry, Shahrood Branch, Islamic Azad University, Shahrood, Iran

    Mohammad Taghi Goodarzi

  3. Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran

    Masoud Homayouni Tabrizi

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The Ethics Committee of Islamic Azad University, Shahrood Branch (Iran) approved our research protocol (IR.IAU.SHAHROOD.REC.1400.011).

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Ghandehari, S., Goodarzi, M.T., Nia, J.I. et al. Evaluation of cytotoxicity, apoptosis, and angiogenesis induced by Kombucha extract-loaded PLGA nanoparticles in human ovarian cancer cell line (A2780). Biomass Conv. Bioref. 13, 13103–13115 (2023). https://doi.org/10.1007/s13399-021-02283-2

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