Deciphering Annona's Anticancer Potential: Comparative Analysis of Crude Extract Versus Nanoformulation on SCC-25 Oral Cancer Cells Through cell Viability, Apoptosis, Cell Cycle, ROS, and MMP Analysis.

Abstract

Annona muricata contains acetogenins, which have shown promising anticancer activity against various cell lines. This study aims to evaluate and compare the anticancer activity of the crude extract of Annona muricata and its nano formulation on Squamous Cell Carcinoma-25 (SCC-25) oral cancer cell lines. The crude extract of Annona muricata was prepared using standard extraction techniques, while its nano formulation was synthesized through nanoparticle fabrication methods. Authenticated SCC-25 cell lines were obtained from ATCC and cultured and treated with varying concentrations of both the crude extract and nano formulation. Cell viability assays, apoptosis assays, Cell Cycle assay, ROS, and MMP analysis techniques were employed to assess the anticancer activity and mechanism of action. In the MTT assay, the Annona formulation treated cells exhibited lower IC50 values compared to the crude extract treated SCC-25 cell lines. In the cell cycle assay, the Annona crude extract inducedhigher cell cycle arrest in the G1 phase in SCC-25 cell lines compared to the control. The nano formulationof Annona demonstrated significantly higher cell cycle arrest in G1 phase compared to both the control and the Annona crude extract-treated SCC-25 cell lines. The crude extract showed less apoptotic activity in apoptosis assay when compared to control, whereas the Annona formulation exhibited higher late apoptosis compared to the control, indicating the potential anticancer properties of Annona. The mean fluorescent intensity test of SCC-25 oral cancer cells treated with Annona crude extract and Annona formulation showed a significant loss of Mitochondrial membrane potential compared to the control. The percentage of MMP was lower in Annona-treated cells, while the Annona formulationtreated cells showed similar results to the control. The mean fluorescent intensity of ROS in SCC-25 oral cancer cells treated with Annona crude extract and Annona formulation showed significantly lower Reactive oxygen species production compared to the control. The percentage of ROS was lower in Annona treated cells compared to the formulation, but the Annona formulation-treated cells showed lower values than the control. In conclusion, both the crude extract and nano formulation of Annona muricata possess potent anticancer activity against SCC-25 oral cancer cell lines. However, the nano formulation exhibited superior efficacy, suggesting its potential for further development as a therapeutic agent for oral cancer treatment.