Formulation of a gambier catechin-loaded nanophytosome and the MTT assay on HeLa cell lines
DOI:
https://doi.org/10.46542/pe.2023.232.1924Keywords:
Gambier catechin, Nanophytosome, MTT assayAbstract
Background: Catechins are good free radical scavengers but exhibit low stability and permeability. Nanophytosomes are currently being developed as the delivery system for phytoconstituents to protect them from decomposition by oxidants or enzymes and increase their permeability.
Objectives: To formulate gambier catechin-loaded nanophytosomes and perform the MTT assay on HeLa cell lines.
Method:Five formulations were prepared using soya lecithin at various molar ratios of cholesterol by thin-layer hydration and sonication. The nanophytosomes were characterised by the determination of vesicle size, zeta potential, polydispersity index, morphology with Transmission Electron Microscope, Fourier Transform Infrared Spectroscopy (FT-IR) analysis, freeze and thaw test, entrapment efficiency, and the in vitro cytotoxicity test.
Result: The optimal formula (F4) with a molar ratio of 1:1:0.8 (catechin:lecithin:cholesterol) resulted in spherical vesicles with an average size (106 ± 0.218) nm, zeta potential -68 mV, polydispersity index 0.412, 93.5% entrapment efficiency and that were stable to temperature changes. FT-IR showed the formation of catechins and lecithin complexes. The activity of catechin-loaded nanophytosomes against HeLa cells showed an IC50 of 36.307 g/ml. There was a significant difference in the average percentage of cells undergoing apoptosis in all treatment groups (p < 0.05).
Conclusion: Catechin-loaded nanophytosomes with a molar ratio of 1:1:0.8 (catechin:lecithin:cholesterol) showed moderate cytotoxic activity against HeLa cell lines.
References
Alshatwi, A. A. (2010). Catechin hydrate suppresses MCF-7 proliferation through TP53/Caspase-mediated apoptosis. Journal of Experimental and Clinical Cancer Research, 29(1). https://doi.org/10.1186/1756-9966-29-167
Babazadeh, A., Zeinali, M., & Hamishehkar, H. (2017). Nano-Phytosome: A Developing Platform for Herbal Anti-Cancer Agents in Cancer Therapy. Current Drug Targets, 18(999). https://doi.org/10.2174/1389450118666170508095250
Chakrabarty, S., Ganguli, A., Das, A., Nag, D., & Chakrabarti, G. (2015). Epigallocatechin-3-gallate shows anti-proliferative activity in HeLa cells targeting tubulin-microtubule equilibrium. Chemico-Biological Interactions, 242. https://doi.org/10.1016/j.cbi.2015.11.004
Cheng, Z., Zhang, Z., Han, Y., Wang, J., Wang, Y., Chen, X., Shao, Y., Cheng, Y., Zhou, W., Lu, X., & Wu, Z. (2020). A review on anti-cancer effect of green tea catechins. In Journal of Functional Foods (Vol. 74). https://doi.org/10.1016/j.jff.2020.104172
Hara-Terawaki, A., Takagaki, A., Kobayashi, H., & Nanjo, F. (2017). Inhibitory activity of catechin metabolites produced by intestinal microbiota on proliferation of HeLa cells. Biological and Pharmaceutical Bulletin, 40(8). https://doi.org/10.1248/bpb.b17-00127
Hebbar, S., & Mathias, A. C. (2018). Phytosomes: A Novel Molecular Nano Complex Between Phytomolecule and Phospholipid as a Value added Herbal Drug Delivery System. Article in International Journal of Pharmaceutical Sciences Review and Research, 51(1)
Hosen, N. (2017). Profil Sistem Usaha Pertanian Gambir di Sumatera Barat. Jurnal Penelitian Pertanian Terapan, 17(2). https://doi.org/10.25181/jppt.v17i2.291
Hussain, H., Santhana Raj, L., Ahmad, S., Abd. Razak, M. F., Wan Mohamud, W. N., Bakar, J., & Ghazali, H. M. (2019). Determination of cell viability using acridine orange/propidium iodide dual-spectrofluorometry assay. Cogent Food and Agriculture, 5(1). https://doi.org/10.1080/23311932.2019.1582398
Kassim, M. J., Hussin, M. H., Achmad, A., Dahon, N. H., Suan, T. K., & Hamdan, H. S. (2011). Determination of total phenol , condensed tannin and flavonoid contents and antioxidant activity of Uncaria gambir extracts. 22(1), 50–59
Kumar, A., & Dixit, C. K. (2017). Methods for characterization of nanoparticles. In Advances in Nanomedicine for the Delivery of Therapeutic Nucleic Acids. https://doi.org/10.1016/B978-0-08-100557-6.00003-1
Marlinda. (2018). Identifikasi Kadar Katekin pada Gambir (Uncaria gambier Roxb.). Jurnal Optimalisasi, 4(1)
Nakayama, M., Shimatani, K., Ozawa, T., Shigemune, N., Tsugukuni, T., Tomiyama, D., Kurahachi, M., Nonaka, A., & Miyamoto, T. (2013). A study of the antibacterial mechanism of catechins: Isolation and identification of Escherichia coli cell surface proteins that interact with epigallocatechin gallate. Food Control, 33(2). https://doi.org/10.1016/j.foodcont.2013.03.016
Ochi, M. M., Amoabediny, G., Rezayat, S. M., Akbarzadeh, A., & Ebrahimi, B. (2016). In vitro co-delivery evaluation of novel pegylated nano-liposomal herbal drugs of silibinin and glycyrrhizic acid (Nano-phytosome) to hepatocellular carcinoma cells. Cell Journal, 18(2)
Purnamasari, N. A. D., Dzakwan, M., Pramukantoro, G. E., Mauludin, R., & Elfahmi. (2020). Evaluation of myricetin nanophytosome with thin-sonication layer hydration method using ethanol and acetone solvents. International Journal of Applied Pharmaceutics, 12(5). https://doi.org/10.22159/ijap.2020v12i5.36520
Yamauchi, R., Sasaki, K., & Yoshida, K. (2009). Identification of epigallocatechin-3-gallate in green tea polyphenols as a potent inducer of p53-dependent apoptosis in the human lung cancer cell line A549. Toxicology in Vitro, 23(5). https://doi.org/10.1016/j.tiv.2009.04.011
