Development and characterization of Zingiber zerumbet rhizome crude extract-loaded nanoemulsions: In vitro release and cytotoxicity assessment against MDA-MB-231 breast cancer cell line

Zingiber zerumbet, a rhizome from the ginger family (Zingiberaceae), possesses significant therapeutic potential due to its bioactive compounds, particularly the lipophilic component zerumbone. Zerumbone is reported to inhibit breast cancer growth but is limited by poor solubility and low absorption. However, improving these properties could enhance its potential as a cancer treatment. This research focuses on developing a nanoemulsion incorporating the rhizome of Zingiber zerumbet crude extract (ZZCE) for use in breast cancer therapy. The ZZCE was prepared using the maceration method and characterized through Gas Chromatography-Mass Spectrometry (GC–MS), Fourier Transform Infrared Spectroscopy (FTIR), and UV–visible spectroscopy (UV–Vis). GC–MS and FT-IR analyses confirmed the presence of zerumbone and its functional groups, while UV-visible spectroscopy indicated a maximum absorption at 285 nm. A nanoemulsion was prepared using a low-energy emulsification method. The composition of the nanoemulsions was determined based on the ternary phase diagram. Four nanoemulsion formulations (F3, F6, F11, and F12) demonstrated excellent stability over six months of storage at room temperature, with droplet sizes of 196.5, 214.1, 211.2, and 83.6 nm, respectively. Due to its small droplet size, formulation F12 was selected for further testing. In vitro release analysis of F12 showed a sustained release pattern with 35.9 % cumulative release over 24 h, supporting its potential as a controlled delivery vehicle. A cytotoxicity study against the MDA-MB-231 breast cancer cell line revealed that the crude extract had an IC₅₀ value of 14.25 μg/mL, indicating strong cytotoxic activity. Although the ZZCE-loaded nanoemulsion did not exhibit in vitro cytotoxicity up to 100 µg/mL, its physicochemical stability, small particle size, and sustained release profile suggest its potential as a nano-delivery platform. Further optimization and mechanistic studies are needed to evaluate its suitability for anticancer applications.

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