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Annona muricata L. leaves extract loaded on chitosan nanoparticles combined with methotrexate induce Cytotoxicity in A549 cancer cell line

Article scientifique 2023 Anglais

Résumé

Abstract Annona muricata Linn (AM) and methotrexate (MTX)-loaded chitosan (Ch) as a natural cationic, bio-degradable and biocompatible polymer were investigated for their genotoxic and cytotoxic effects in their normal and nano form. The AM-ChNP and MTX-ChNP were analyzed and characterized according to fourier transform infrared spectroscopy (FTIR), particle size, Zeta potential, transmission electron microscope (TEM) and encapsulation efficiency. The chitosan concentration (3:1) was found to be an important parameter for optimizing the dispersion of AM-ChNPs and of MTX-ChNPs nanocomposites over long time. The efficiency was found to be 87.3% and size of 100–200 nm. The stabilty of the nanocomposites offers a good sign in cancer cell line treatment. Seven groups were prepared using the A549 lung cell line: control group, A. muricata leaf extract (AM), Methotrexate (MTX), AM-ChNPs, MTX-ChNPs, AM + MTX, and AM-ChNPs + MTX-ChNPs. 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide ( MTT) assay for testing cell cytotoxicity; gene expression of Caspase- 3; Bcl- 2; TNF- α; and Cyclin A genes. Cell cycle progress and the percentages of apoptosis and necrosis additionally were assessed. The combination of AM-ChNPs and MTX-ChNPs revealed clear action against A549 cancer cell line. The IC50 values for AM-ChNPs + MTX-ChNPs nano-group compared to AM + MTX group increased by 1.7 folds. Moreover, the apoptotic Caspase-3 gene expression was significantly upregulated by 8 folds. However, antiapoptotic Bcl-2 gene expression was unsignificantly downregulated by 0.16 folds. The inflammatory TNF-α gene expression was significantly upregulated by 6 folds, and the Cyclin A gene expression was downregulated by 0.29 folds, in the combination group of AM-ChNPs + MTX-ChNPs. An increase was discovered through flow cytometric analysis in the G2/M phase cell population driven into apoptosis. This study highlights on the combined effects on genotoxicity and cytotoxicity for both AM-ChNPs and MTX-ChNPs and their free forms; as well as their apoptogenic activities.

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Muhammad, N., Elbeltagy, R., Diab, M., Gardouh, A., El-Habit, O. (2023). Annona muricata L. leaves extract loaded on chitosan nanoparticles combined with methotrexate induce Cytotoxicity in A549 cancer cell line. https://doi.org/10.21203/rs.3.rs-2992634/v1

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Auteur(s)

Nada A.E. Muhammad Rasha Salah Elbeltagy Marwa S.M. Diab Ahmed R. Gardouh Ola H. El-Habit

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