Iron oxide nanoparticles coated with Glucose and conjugated with Safranal (Fe3O4@Glu-Safranal NPs) inducing apoptosis in liver cancer cell line (HepG2)

Abstract

Magnetic nanoparticles can be considered a reliable tool for targeted drug delivery to cancer tissues. Based on this, in this study, the anticancer effect of iron oxide nanoparticles coated with glucose and conjugated with Safranal (Fe₃O₄@Glu-Safranal NPs) on a liver cancer cell line (HepG2) was investigated. Physicochemical properties of nanoparticles were characterized using FT-IR, XRD, VSM, EDS-mapping, SEM and TEM imaging, zeta potential, and DLS analyses. MTT test was used to investigate the inhibitory effect of nanoparticles on cancer and normal cell lines. Also, the reactive oxygen species (ROS) level, the population of apoptotic cells, and cell cycle analysis were evaluated in control and nanoparticle-treated cells. The synthesized particles were spherical, in a size range of 17-49 nm, without impurities, with a surface charge of - 13 mV and hydrodynamic size of 129 nm, and with magnetic saturation of 22.5 emu/g. The 50% inhibitory concentration (IC₅₀) of Safranal, Fe₃O₄, Fe₃O₄@Glu-Safranal and Cisplatin drug on liver cancer cells were 474, 1546, 305 and 135 µg/mL, respectively. While, the IC₅₀ of Fe₃O₄@Glu-Safranal for normal cell line was 680 µg/mL. Treating liver cancer cells with nanoparticles significantly increased the population of apoptotic cells from 2.5% to 34.7%. Furthermore, the population of the cells arrested at the G2/M phase increased in nanoparticle-treated cells. Due to the biocompatibility of the constituent compounds of these nanoparticles, their magnetic properties, and their inhibitory effects on cancer cells, Fe₃O₄@Glu-Safranal NPs can be further considered as a promising anticancer compound.

Keywords: Apoptosis; Drug delivery; Fe3O4@Glu-Safranal; Liver cancer; Safranal.

Fig. 1

FT-IR spectrogram of Fe₃O₄@Glu-Safranal, Safranal, and Fe₃O₄ nanoparticles

Fig. 2

a XRD and b XPS analyses of Fe₃O₄@Glu-Safranal nanoparticles. c Fe 2p spectrum

Fig. 3

a TEM, b SEM and c particle size distribution of Fe₃O₄@Glu-Safranal nanoparticles. The synthesized particles were spherical and in a size range of 17–49 nm

Fig. 4

a Zeta potential, and b DLS analysis of Fe₃O₄@Glu-Safranal nanoparticles. Surface charge of the nanoparticles was − 13 mV and hydrodynamic size was 129 nm

Fig. 5

Elemental mapping of Fe₃O₄@Glu-Safranal nanoparticles. The particles contained C, Fe, and O atoms, and no elemental impurity was observed

Fig. 6

Magnetic saturation curve of Fe₃O₄@Glu-Safranal nanoparticles. The particles had magnetic properties and the maximum magnetic property was 22.5 emu/g which was observed at 6000Oe

Fig. 7

MTT assay showed that the IC₅₀ of Safranal, Fe₃O₄, Fe₃O₄@Glu-Safranal and Cisplatin on liver cancer cells were 474, 1546, 305 and 135 µg/mL, respectively. The IC₅₀ of Fe₃O₄@Glu-Safranal on normal cell line was 680 µg/mL

Fig. 8

ROS generation in a control and b Fe₃O₄@Glu-Safranal treated cells. Treating cancer cells with the nanoparticles resulted in a considerably higher ROS generation

Fig. 9

Flow cytometry analysis a control, b Fe₃O₄@Glu-Safranal NPs, c Cisplatin drug and d Fe₃O₄ NPs treated cancer cells. Treating with the Fe₃O₄@Glu-Safranal NPs and Cisplatin drug considerably increased the population of apoptotic cells. Q1: necrotic cells, Q2: late apoptosis, Q3: primary apoptosis, and Q4: live cells

Fig. 10

Cell cycle analysis of a control and b Fe₃O₄@Glu-Safranal treated cells showed an increased population of the cells arrested in G2/M Phase