Assessment of the Cytotoxic and Apoptotic Impacts of Bimetallic Zinc Iron Oxide Nanoparticles on HuH-7 Cell Line

Journal Article

2026

Published in: https://doi.org/10.2147/DDDT.S623623

Background and Observation: One of the main causes of death worldwide is hepatic carcinoma, which is resistant to several

anticancer drugs. This work investigated the cytotoxic effects of bimetallic zinc iron oxide ZnFe2O4 nanoparticles (ZnFe2O4NPs) on

the human liver cancer (HuH-7) cell line and demonstrated the minimally toxic doses of these particles.

Methods: Before being exposed to target cell line, ZnFe2O4NPswere examined using transmission electron microscopy (TEM),

scanning electron microscopy (SEM), and EDX. ZnFe2O4NPs showed a spherical form and a size of 29.50 ±1.5 nm. MTT and NRU

tests were used to assess the cytotoxic impact of different concentrations (0, 10,30, 60, 120, 150 and 200 µg/mL) of ZnFe2O4NPs on

HuH-7 cell line. HuH-7 cell line showed a significant lethal effect at the highest concentration of NP exposure, and the cytotoxicity of

NPs increased in a concentration-dependent way.

Results: The median inhibitory concentration (IC50) for HuH-7 cell line at 24 hours was found to be 61 μg/mL of NPs based on the

MTT results. At higher ZnFe2O4NPs concentrations, intracellular ROS levels were increased. At high ZnFe2O4NPs concentrations, HuH-

7 cell line showed decreased levels of catalase and glutathione (GSH). Rhodamine 123 labeling was used to measure the reduction in

mitochondrial membrane potential in both control and NP-exposed cell line. Contrary to their effectiveness at low concentrations,

exposure to high doses of ZnFe2O4NPs inhibited cell growth, proliferation, and migration in a wound healing assay. While intended to

accelerate healing, high NP concentrations (30.5 µg/mL, depending on type and size) can cause significant cytotoxicity to human liver

cancer cell line. Maximum apoptotic cell line were seen in HuH-7 cell line at high NP concentrations. At higher NP concentrations,

HuH-7 cell line showed an increase in caspase-3 activity.

Conclusion: The aforementioned findings point out the significance of ZnFe2O4NPs as a cytotoxic agent caused by oxidative stress

that could be useful in various cancer treatments.