Encapsulation of Artemisia scoparia extract in chitosan-myristate nanogel with enhanced cytotoxicity and apoptosis against hepatocellular carcinoma cell line (Huh-7)

Abstract

In the current study, encapsulation by chitosan (CS)-myristic acid (MA) nanogel was used to enhance the cytotoxic and apoptosis activities of Artemisia scoparia extract against Hepato-carcinoma cell line (Huh-7). The synthesized CS-MA nanogel had uniform size distribution below 100 nm with the spherical structure. Subsequently, cytotoxicity of free A. scopari extract, CS-MA nanogel and CS-MA nanogel-encapsulated extract was evaluated on Huh-7 cell line using MTT (3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Finally, the apoptotic effects of CS-MA nanogel-encapsulated extract was evaluated using Real-time PCR and flow cytometry techniques. The results of MTT assay showed that the CS-MA nanogel-encapsulated extract was more effective against Huh-7 cell line comparing to the free extract. In addition, CS-MA nanogel-encapsulated extract induced the apoptosis in Huh-7 cell line with up-regulation of Bax, and miR192 genes along with the suppression of Bcl-2, MMP2, MMP9 and cyclin D1 (P < 0.01). Moreover, the flow cytometric analysis of cell cycle distribution of Huh-7 cells showed enhanced sub-G1 peaks by the CS-MA nanogel-encapsulated extract treatment, which is indicative of apoptosis pathway. In conclusion, A. scoparia extract in the form of CS-MA nanogel-encapsulated can be used as an effective tool to enhance cytotoxicity against Huh-7 cell line and could be a promising candidate for hepato-carcinoma therapy.