Synthesis and Characterization of Liposomal Nanocarriers Containing Pomegranate Flower Extract to Investigate Its Toxicity on the MCF-7 Breast Cancer Cell Line
Subject Areas : Journal of Animal Biology
Ayda Shafigh
1
,
Lida Eftekharivash
2
*
,
Ali Akbar Seifi Ashkezari
3
1 - Department of Microbiology, Maragheh Brench, Islamic Azad University, Maragheh, Iran
2 - Department of Microbiology, Maragheh Brench, Islamic Azad University, Maragheh, Iran
3 - Medical Student, Iran University of Medical Sciences, Tehran, Iran
Keywords: Breast cancer, Pomegranate flower extract, Liposome, MCF-7, MTT,
Abstract :
Although pomegranate flower extract has anticancer properties, it requires carriers to improve its efficacy by delivering it to the target cells. Therefore, this study focuses on the development of pomegranate flower extract-loaded nanoliposomes and investigates their effects on MCF-7 breast cancer cells. In this study, after extracting the pomegranate flower extract and determining properties such as the maximum wavelength and standard curves, the extract-loaded nanoliposomes were synthesized using the thin-film method. The physicochemical characteristics of the nanoliposomes, including encapsulation efficiency, drug release, size, and zeta potential, were evaluated using spectrophotometry and DLS, while their morphology was studied using Fe-SEM and AFM microscopy. Finally, the antitumor activity of the nanoliposomes containing the extract and the free extract was assessed on MCF-7 cancer cells using the MTT assay. The results showed that the encapsulation efficiency of the synthesized nanostructure was 73.2 ± 2.6%, and the maximum release of the extract under normal and cancerous cell conditions was 51.3 ± 1.4% and 67.2 ± 2.6%, respectively. The nanoparticle size was 155.7 ± 4.1 nm, their polydispersity index was 0.345, and the zeta potential was -17.1 ± 3.4 mV. Microscopic images indicated that the nanoparticles had a spherical and uniform shape with no aggregation or precipitation. The nanoliposomes containing the extract showed higher toxicity on MCF-7 cells compared to the free extract, exhibiting a lower IC50, while the blank system showed minimal toxicity to healthy fibroblast cells. The nanostructure developed in this study not only demonstrated favorable physicochemical properties but also effectively loaded the pomegranate flower extract and significantly enhanced its antiproliferative effects, making it a promising candidate for breast cancer research.
1. Akbari, P., Taebpour M., Akhlaghi M., Hasan S.H., Shahriyari S., Parsaeian M., Haghirosadat B.F., Rahdar A., Pandey S. 2024. Regulation of the P53 tumor suppressor gene and the Mcl-2 oncogene expression by an active herbal component delivered through a smart thermo-pH-sensitive PLGA carrier to improve osteosarcoma treatment. Medical Oncology, 41(3):68.
2. Akhlaghi, M., Eftekharivash, L., Parnian, F., Taebpour, M., Rostamian, T., Zarezadeh Mehrizi, M., Haghirosadat, B.F. 2021. Synthesis and study of cellular and physicochemical properties of nanoliposomes containing aqueous extract of Hedera Helix. New Cellular and Molecular Biotechnology Journal, 12(45):97-112.
3. Akhlaghi, M., Eftekharivash, L., Taebpour, M., Afereydoon, S., Ebrahimpour, M., Mehrizi, M.Z., fatemeh Haghirosadat, B. 2022. Improving the therapeutic performance of glycyrrhiza glabra hydroalcoholic extract using liposomal nano-carriers and their characterization. Disease and Diagnosis, 11(2):39-48.
4. Akhlaghi, M., Taebpour, M., Lotfabadi, N. N., Naghib, S. M., Jalili, N., Farahmand, L., Tofighi, D. 2022. Synthesis and characterization of smart stimuli-responsive herbal drug-encapsulated nanoniosome particles for efficient treatment of breast cancer. Nanotechnology Reviews, 11(1):1364-1385.
5. American cancer society. 2025. cancer facts and figures 2025. Atlanta american cancer.
6. Baghiani, M., Majdizadeh, M., Haghiralsadat, B.F. 2019. Experimental study: experimental evaluation of phospholipid system containing doxorubicin HCL for use in chemotherapy. Journal of Shahid Sadoughi University of Medical Sciences, 26(11):945-956.
7. Bahrami-Banan, F., Sheikhha, M.H., Ghasemi, N., Majdizadeh, M., Haghiralsadat, B.F. 2018. Preparation and study of nano-niosomes containing doxorubicin and evaluation of its toxicity on acute myeloblastic leukemia cell line KG-1. Journal of Payavard Salamat, 12(4):309-323.
8. Deng, Y., Li, Y., Yang, F., Zeng, A., Yang, S., Luo, Y., Yin, W. 2017. The extract from Punica granatum (pomegranate) peel induces apoptosis and impairs metastasis in prostate cancer cells. Biomedicine and Pharmacotherapy, 93:976-984.
9. DeSantis, C.E., Ma, J., Gaudet, M.M., Newman, L. A., Miller, K. D., Goding Sauer, A., Siegel, R.L. 2019. Breast cancer statistics. CA: A Cancer Journal for Clinicians, 69(6):438-451.
10. DeSantis, C.E., Ma, J., Goding Sauer, A., Newman, L.A., Jemal, A. 2017. Breast cancer statistics, 2017, racial disparity in mortality by state. CA: A Cancer Journal for Clinicians, 67(6):439-448.
11. Eftekhari-Vash, L., Majdizadeh, M. 2024. Fabrication and physicochemical characterization of liposome systems containing Glycyrrhiza glabra hydroalcoholic extract and evaluation of toxicity of free and liposomal extract on healthy human fibroblast cell line (HFF) and MCF-7 breast cancer cell line. Journal of Knowledge and Health in Basic Medical Sciences, 18(4):59-69.
12. Faria, A., Calhau, C. 2011. The bioactivity of pomegranate: impact on health and disease. Critical Reviews in Food Science and Nutrition, 51(7):626-634.
13. Filipczak, N., Pan, J., Yalamarty, S.S.K., Torchilin, V.P. 2020. Recent advancements in liposome technology. Advanced Drug Delivery Reviews, 156:4-22.
14. Ghafari, M., Haghiralsadat, F., Khanamani Falahati‐pour, S., Zavar Reza, J. 2020. Development of a novel liposomal nanoparticle formulation of cisplatin to breast cancer therapy. Journal of Cellular Biochemistry, 121(7):3584-3592.
15. Kirby, C. J., Gregoriadis, G. 2019. A simple procedure for preparing liposomes capable of high encapsulation efficiency under mild conditions. In Liposome Technology (pp. 19–27). CRC Press.
16. Ghashghaei, M., Akhlaghi, M. 2021. Investigation of nanoniosomal formulation containing doxorubicin effect on ovarian cancer cell line (OVCAR-3 cell line). New Cellular and Molecular Biotechnology Journal, 11(43):46-62.
17. Harashima, H., et al. 1994. Enhanced hepatic uptake of liposomes through complement activation depending on the size of liposomes. Pharmaceutical Research, 11:402-406.
18. Kamińska, M., Ciszewski, T., Łopacka-Szatan, K., Miotła, P., Starosławska, E. 2015. Breast cancer risk factors. Menopause Review/Przegląd Menopauzalny, 14(3):196-202.
19. Karimi-Moghddam, A., Nikounahad-Lotfabadi, N., Haghiralsadat, B.F., Majdizadeh, M. 2019. Investigating the effect of lipid nanoparticles containing silibinin anti-cancer drug on the growth of breast cancer MCF-7 cell line. Journal of Torbat Heydariyeh University of Medical Sciences, 6(4):1-12.
20. Li, K., Xiao K., Zhu S., Wang Y., Wang W. 2022. Chinese herbal medicine for primary liver cancer therapy: Perspectives and challenges. Frontiers in Pharmacology, 13:889799.
21. Lindner, L.H., Hossann, M. 2010. Factors affecting drug release from liposomes. Current Opinion in Drug Discovery and Development, 13(1):111-123.
22. Majdizadeh, M., Akbarzadeh, S., Al-Turnachy, H.H., Hemati, M., Akhlaghi, M., Haghiralsadat, B.F., Oroojalian, F. 2025. Synthesis of nano-liposomes containing Mentha piperita essential oil and investigating their stability, antimicrobial, antioxidant, and antiproliferative properties. Nanomedicine Journal, 12(1):122-139.
23. Majdizadeh, M., Rezaei-Zarchi, S., Movahedpour, A.A., Shahi Malmir, H., Sasani, E., Haghiralsadat, B.F. 2018. A new strategy in improving therapeutic indexes of medicinal herbs: Preparation and characterization of nano-liposomes containing Mentha piperita essential oil. SSU Journals, 25(11):853-864.
24. Mirzaei, F., Majdizadeh, M., Fatahi-Bafghi, A., Ehsani, R., Haghiralsadat, B. F. 2021. Fabrication and characterization of liposomal nano-carriers containing essential oils of Trachyspermum ammi to counteract Trichomonas vaginalis. Koomesh, 23(2):283-290.
25. Moghadam, R.N., Majdizadeh, M., Golbashy, M., Haghiralsadat, F., Hemati, M. 2024. Laboratory study: Synthesis and optimization of nano-niosomes containing Bunium persicum essential oil and investigating its toxicity on Trichomonas vaginalis parasite and HFF cell line. Heliyon, 10(16):e35967.
26. Mohseni, M., et al. (2021). A new therapeutic approach for the treatment of breast cancer using synthesis of liposomes containing silybinin and their characterization. New Cellular and Molecular Biotechnology Journal, 11(44):57-70.
27. Momenimovahed, Z., Salehiniya, H. (2019). Epidemiological characteristics of and risk factors for breast cancer in the world. Breast Cancer: Targets and Therapy, 11:151-164.
28. Nagayasu, A., Uchiyama, K., Kiwada, H. 1999. The size of liposomes: A factor that affects their targeting efficiency to tumors and the therapeutic activity of liposomal antitumor drugs. Advanced Drug Delivery Reviews, 40(1–2):75-87.
29. Oun, R., Moussa, Y.E., Wheate, N.J. 2018. The side effects of platinum-based chemotherapy drugs: a review for chemists. Dalton Transactions, 47(19): 6645-6653.
30. Pande, S. 2023. Liposomes for drug delivery: Review of vesicular composition, factors affecting drug release, and drug loading in liposomes. Artificial Cells, Nanomedicine, and Biotechnology, 51(1):428-440.
31. Parnian, F., Hekmati-Moghadam, S.H., Majdizadeh, M., Jebali, A., Haghiralsadat, B.F. 2020. Fabrication of niosomal nano-carriers containing aqueous extract of hedera helix and comparison of toxicity of free extract and niosome extract on HT29 colorectal cancer cell line. Journal of Knowledge and Health in Basic Medical Sciences, 15(3):31-45.
32. Pearce, A., Haas, M., Viney, R., Pearson, S. A., Haywood, P., Brown, C., Ward, R. 2017. Incidence and severity of self-reported chemotherapy side effects in routine care: A prospective cohort study. PloS one, 12(10):e0184360.
33. Pornsunthorntawee, O., Chavadej, S., Rujiravanit, R. 2011. Characterization and encapsulation efficiency of rhamnolipid vesicles with cholesterol addition. Journal of Bioscience and Bioengineering, 112(1):102-106.
34. Rabani, E., Behzadi, R., Majdizadeh, M., Haghiralsadat, B.F. 2022. Fabrication of liposomal formulation containing paclitaxel and comparison of its toxicity with non-liposomal paclitaxel on MCF-7 breast cancer cell line. Pars Journal of Medical Sciences, 18(2):26-40.
35. Sasani, E., Shahi Malmir, H., Daneshmand, F., Majdizadeh, M., Haghiralsadat, B.F. 2020. Synthesis and physiochemical characterization of liponiosomal hybrid nano-carriers as carriers for doxorubicin HCl anti-cancer drug. Journal of Sabzevar University of Medical Sciences, 27(1):35-47.
36. Sasani, E., Shahi Malmir, H., Daneshmand, F., Majdizadeh, M., Haghiralsadat, B.F. 2018. A new study on synthesis and optimization of PEGylated liponiosomal nanocarriers containing curcumin for use in cancer chemotherapy. SSU Journals, 26(6):528-541.
37. Shahi Malmir, H., Kalantar, S.M., Sasani, E., Asgari, M., Majdizadeh, M., Haghiralsadat, B.F. 2019. Synthesis and optimization of niosomal carriers containing doxorubicin in order to achieve a final formulation with high potential in cancer cells’ temperature and acidity. Journal of Shahid Sadoughi University of Medical Sciences, 26(10):879-894.
38. Sharma, P., McClees, S.F., Afaq, F. 2017. Pomegranate for prevention and treatment of cancer: an update. Molecules, 22(1):177.
39. Siyadatpanah, A., Norouzi, R., Mirzaei, F., Haghirosadat, B.F., Nissapatorn, V., Mitsuwan, W., Nawaz, M., Lourdes, M., Hosseini, S. A., Montazeri, M., Majdizadeh, M., Almeida, R. S., Hemati, M., Wilairatana, P., Douglas, H. 2022. Green synthesis of nano-liposomes containing Bunium persicum and Trachyspermum ammi essential oils against Trichomonas vaginalis. Journal of Microbiology, Immunology and Infection, 56(1):150-162.
40. Smith, M.C., Crist R.M., Clogston J.D., McNeil S.E., 2017. Zeta potential: A case study of cationic, anionic, and neutral liposomes. Analytical and Bioanalytical Chemistry, 409:5779-5787.
41. Sun, Y.S., Zhao, Z., Yang, Z.N., Xu, F., Lu, H.J., Zhu, Z.Y., Zhu, H.P. 2017. Risk factors and preventions of breast cancer. International Journal of Biological Sciences, 13(11):1387.
42. Taebpour, M., Akhlaghi, M., Shahriary, S., Hajihosseini, S., Haghiralsadat, F., Oroojalian, F., Tofighi, D. 2022. Synthesis, physicochemical characterization and pharmaceutical function of niosomal nanoparticles-encapsulated bioactive compound for osteosarcoma treatment. Nanomedicine Journal, 9(3):205-215.
43. Taebpour, M., Majdizadeh, M., Haghiralsadat, B.F. 2021. Fabrication and characterization of liposomal nanoparticles containing hydroalcoholic extract of Artemisia absintium and its toxicity on MCF-7 breast cancer cell line. Iranian Journal of Breast Diseases, 14(1):64-77.
44. Taebpour, M., Majdizadeh, M., Haghiralsadat, B.F., Akhlaghi, M. 2021. Fabrication of liposomal nanoparticles containing hydroalcoholic extract of Silybum marianum and evaluation of its toxicity on human osteosarcoma cancer cell line (SAOS-2) and evaluation effect of free hydroalcoholic extract of Silybum marianum and empty liposomes on healthy human fibroblast cell (HFF). Journal of Knowledge and Health in Basic Medical Sciences, 16(1):61-73.
45. 45. Taebpour, M., Majdizadeh, M., Nodoushan, F.S., Akhlaghi, M., Haghiralsadat, B.F. 2023. Evaluation and Comparison of Punica Granatum Peel and Flower Extract Toxicity on MCF-7 Breast Cancer Cell Line and Comparison of Antioxidant Activity of Punica Granatum Peel and Flower Extract. Journal of Knowledge and Health in Basic Medical Sciences, 18(1):9-16.
46. Talei-Ardakani, N., Daneshmand, F., Mirhoseini, M., Majdizadeh, M., Haghiralsadat, B.F. 2020. Fabrication and characterization of physicochemical niosomal nanocarriers containing quercetin flavonoids for therapeutic purposes. Journal of Knowledge and Health in Basic Medical Sciences, 15(1):fa32-fa40
47. van den Boogaard, W.M., Komninos, D.S., Vermeij, W.P. 2022. Chemotherapy side-effects: not all DNA damage is equal. Cancers, 14(3):627.
48. Waks, A.G., Winer, E.P. 2019. Breast cancer treatment: a review. Jama, 321(3):288-300.
49. Zarehzadeh-Mehrizi, M., Majdizadeh, M., Zarezadeh-Mehrizi, M., Afshoun, A., Zhian-Mehrizi, M., Haghiralsadat, B.F. 2023. Investigating the performance of liposomal formulations containing clove essential oil made by thin layer and heat method in temperature and pH conditions similar to the mouth in order to be used in the research of oral and dental diseases. Experimental Animal Biology, 12(46):15-26.