Synthesis and Characterization of Q0-Eugenol Nanoemulsion for Drug Delivery to Breast and Hepatocellular Cancer Cell Lines

Abstract

Cancer is the second cause of death globally. The advent of nanoparticles has revolutionized drug delivery in cancer treatment. This study introduces a novel Q0-eugenol nanoemulsion designed for enhanced drug delivery. Synthesis of nanoemulsion was performed by the homogenization technique and ultrasonication for size reduction. DLS, FTIR, and UV-vis spectroscopy were used for characterization of Q0-eugenol nanoemulsion. MTT assay and flow cytometry assay were applied to determine the fibroblast, KPL1, and PLC/PRF/5 cell lines to consider the cell viability and apoptosis percentage, respectively. The oxidative status of Q0-eugenol nanoemulsion was evaluated by the ferric reducing antioxidant power assay and the thiobarbituric acid reactive substance assay. The zeta potential charge of eugenol and Q0-eugenol nanoemulsion formulation was equal to 45.1 and 22.7 mV, respectively. Eugenol nanoemulsion and Q0-eugenol nanoemulsion treatment exhibited antiproliferative effect against breast and liver cancer cell lines (p < 0.05). Eugenol nanoemulsion and Q0-eugenol nanoemulsion induced apoptosis through reactive oxygen species-dependent mechanism. Apoptosis rates for PLC/PRF/5 treated with eugenol nanoemulsion, Q0, and Q0-eugenol nanoemulsion were 26.7, 84.7, and 16.8%, respectively. Apoptosis rates for fibroblasts treated with eugenol nanoemulsion, Q0, and Q0-eugenol nanoemulsion were 22.6, 37.8, and 24.4%, respectively. Moreover, in the KPL1 cell line, the apoptosis rates were reported as 11.1, 57.9, and 19% with eugenol nanoemulsion, Q0, and Q0-eugenol nanoemulsion treatments, respectively. Eugenol nanoemulsion and Q0-eugenol nanoemulsion exhibited similar antiproliferative mechanisms against both cell lines. In conclusion, our results showed eugenol nanoemulsion and Q0-eugenol nanoemulsion as a potential treatment option against hepatocellular and breast cancer via induced ROS and apoptosis.

1

Size distribution of the eugenol nanoemulsion (A) and Q0-eugenol nanoemulsion (B) by DLS.

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(A) Photo of Q0-eugenol nanoemulsion. (B) TEM of Q0-eugenol nanoemulsion. (C) Size distribution histogram of Q0-eugenol nanoemulsion.

3

FTIR of (A) Q0, (B) eugenol nanoemulsion, and (C) Q0-eugenol nanoemulsion.

4

UV–vis diagram of Q0 (blue), eugenol nanoemulsion (red), and Q0-eugenol nanoemulsion (green).

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PLC/PRF/5 hepatocellular cancer cell line was treated with Q0, eugenol nanoemulsion, and Q0-eugenol nanoemulsion; * ≤ 0.05, ** ≤ 0.01, *** ≤ 0.001, **** ≤ 0.0001.

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KPL1 breast cancer cell line was treated with Q0, eugenol nanoemulsion, and Q0-eugenol nanoemulsion; * ≤ 0.05, ** ≤ 0.01, *** ≤ 0.001, **** ≤ 0.0001.

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Fibroblast cell line was treated with Q0, eugenol nanoemulsion, and Q0-eugenol nanoemulsion; * ≤ 0.05, ** ≤ 0.01, *** ≤ 0.001, **** ≤ 0.0001.

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Cell morphology alteration under Q0, eugenol nanoemulsion, and Q0-eugenol nanoemulsion treatment (20×).

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Influence of Q0, eugenol nanoemulsion, and Q0-eugenol nanoemulsion on the scales of apoptosis in fibroblast, KPL1 breast cancer, and PLC/PRF/5 hepatocellular cancer cells.