Augmenting the cytotoxic effect of the aerial parts of Carissa macrocarpa (Eckl.) A. DC. cultivated in Egypt by using surfactant-free nanoemulsion.

Document Type : Original Article

Authors

1 Department of Pharmacognosy, National Research Centre, Cairo, Egypt.

2 Department of Pharmaceutical Technology, National Research Centre, Cairo, Egypt.

3 Department of Food Science and Nutrition, National Research Centre, Cairo, Egypt.

4 Department of Pharmacognosy, Faculty of Pharmacy, Cairo University, Egypt.

Abstract

Carissa macrocarpa (Eckl.) A. DC. was previously reported for its anticancer and cytotoxic activities due to its high phenolic and flavonoid contents. The crude methanolic extract of the aerial parts of Carissa macrocarpa (Eckl.) A. DC cultivated in Egypt proved a potent cytotoxicity against HEPG2 and HCT116. The biologically active extract was encapsulated in a nanocellulose-stabilized nanoemulsion (NC-NE) in a trial to enhance its aqueous solubility and ameliorate its effect while avoiding the use of surfactants. The selected NC-NE had a particle size of 194 ± 18 nm with a uniform dispersion (PDI= 0.2) and a good physical stability (Z = -32 mV). Also, the release enhancement was about 3 fold compared to the crude extract. NC-NE had an almost spherical outline (as detected by TEM). The cytotoxicity of the prepared nanoemulsion was studied to determine the difference in efficacy between it and the crude methanolic extract of the plant under investigation. Percentage of inhibition of the selected nanoformulation (NC-NE) against HePG2 and HCT116 was 100% (IC50 20.62 µg/mL, IC90 33.13 µg/mL) and 99.3% (IC50 32.80 µg/mL, IC90 58.22 µg/mL) respectively at 100 ppm, while that of the methanolic extract (E) was 36.5% (IC50 was more than 100 µg/mL) and 71.3% (IC50 74.11 µg/mL, IC90 117.54 µg/mL) respectively at 100 ppm. It is obviously clear from our results that preparation of nanoformulation (NC-NE) increased the cytotoxicity of carissa macrocarpa crude methanolic extract. Phytochemical investigation of the active crude methanolic extract revealed the presence of many phenolic and flavonoid compounds. The major phenolic compounds were chlorogenic acid, protocatechuic acid and caffeic acid which represented 1024.33 μg/g, 257.48 μg/g and 179.90 μg/g respectively, while the three identified flavonoids were rutin, apigenin-7-glucoside and quercetin that represented 299.22 μg/g, 254.90 μg/g and 16.39 μg/g respectively. These compounds were separated and identified by spectroscopic and chromatographic techniques. As far as we know, it is the first time to study the effect of nanoparticle preparation on cytotoxic effect of Carissa macrocarpa methanolic extract.

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