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. 2024 Jun 6;29(11):2695.
doi: 10.3390/molecules29112695.

Pickering Emulsion of Oleoresin from Dipterocarpus alatus Roxb. ex G. Don and Its Antiproliferation in Colon (HCT116) and Liver (HepG2) Cancer Cells

Piman Pocasap  1 , Kawintra Tamprasit  2   3 , Thanyathanya Rungsri  4 , Karnchanok Kaimuangpak  2 , Tarapong Srisongkram  3   5 , Somporn Katekaew  6 , Khanita Kamwilaisak  7 , Ploenthip Puthongking  5 , Natthida Weerapreeyakul  3   5
Affiliations

Pickering Emulsion of Oleoresin from Dipterocarpus alatus Roxb. ex G. Don and Its Antiproliferation in Colon (HCT116) and Liver (HepG2) Cancer Cells

Piman Pocasap et al. Molecules. .

Abstract

Oleoresin of Dipterocarpus alatus Roxb. ex G. Don (DA) has been traditionally used for local medicinal applications. Several in vitro studies have indicated its pharmacological potential. However, the low water solubility hinders its use and development for pharmaceutical purposes. The study aimed to (1) formulate oil-in-water (o/w) Pickering emulsions of DA oleoresin and (2) demonstrate its activities in cancer cells. The Pickering emulsions were formulated using biocompatible carboxylated cellulose nanocrystal (cCNC) as an emulsifier. The optimized emulsion comprised 3% (F1) and 4% (v/v) (F2) of oleoresin in 1% cCNC and 0.1 M NaCl, which possessed homogeneity and physical stability compared with other formulations with uniform droplet size and low viscosity. The constituent analysis indicated the presence of the biomarker dipterocarpol in both F1 and F2. The pharmacological effects of the two emulsions were demonstrated in vitro against two cancer cell lines, HepG2 and HCT116. Both F1 and F2 suppressed cancer cell viability. The treated cells underwent apoptosis, as demonstrated by distinct nuclear morphological changes in DAPI-stained cells and Annexin V/PI-stained cells detected by flow cytometry. Our study highlights the prospect of Pickering emulsions for oleoresin, emphasizing enhanced stability and potential pharmacological advantages.

Keywords: Dipterocarpus alatus; Pickering emulsion; anticancer activity; cell-based assay; oil-in-water.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Physical appearance of DA oleoresin Pickering emulsion with 1% cCNC (A) and 2% cCNC (B) as emulsifier. The emulsions were formulated by varying the oleoresin ratio in the oil phase (10% total) and NaCl in the water phase (90%). Following preparation and storage (25 °C for 30 days), the physical stability of the emulsions was assessed through visual observation.
Figure 2
Figure 2
The physical changes in Pickering emulsions F1 and F2 during storage at 25 °C and 4 °C for 56 days.
Figure 3
Figure 3
Homogeneity test by image analysis. Homogeneity was assessed via image analysis, comparing the intensity of areas X, Y, and Z among formulations F1 (A), F2 (B), and formulation 1-01-A (used as a positive control) (C). A higher p-value indicates increased uniformity among these areas.
Figure 4
Figure 4
Fluorescence microscopic image of Pickering emulsions F1 (A) and F2 (B) stained with oil red-O dye and their size distribution.
Figure 5
Figure 5
Cell viability in the HepG2 (A) and HCT116 (B) cancer cell lines following treatment with Pickering emulsions F1 and F2 for 24 h. The values represent the mean ± SD of triplicate measurements.
Figure 6
Figure 6
Fluorescent microscopy was conducted on HepG2 (A) and HCT116 (B) cells stained with DAPI after being exposed to Pickering emulsions F1, F2, and cisplatin (positive control) at their respective IC50 concentrations for 24 h. The 1 × IC50 values for F1 and F2 against HepG2 cells were 1.25 and 0.92 mg/mL, respectively. Against HCT116 cells, the 1 × IC50 values for F1 and F2 were 3.45 and 1.77 mg/mL, respectively.
Figure 7
Figure 7
Apoptotic cell death induced by Pickering emulsions F1 and F2. Dot plots illustrate the mode of cell death in HepG2 (A) and HCT116 (B) after 24 h of F1 and F2 treatment. Quadrants 1 (Q1) to 4 (Q4) indicate necrosis, late apoptosis, viable cells, and early apoptosis, respectively. Total apoptotic cell death in HepG2 (C) and HCT116 (D) after F1 and F2 treatment for 24 h was quantified. Values are presented as the mean ± SD of triplicate measurements, and significance was determined at a p-value < 0.05. The 2 × IC50 values for F1 and F2 against HepG2 cells were 2.50 and 1.84 mg/mL, respectively. Against HCT116 cells, the 2 × IC50 values for F1 and F2 were 6.90 and 3.54 mg/mL, respectively.
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