Evaluation of anti- cancer and antioxidant properties of nanoemulsions synthesized by Nigella Sativa L. tincture

Document Type : Research Paper


Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran


Objective(s): Today, the use of medicinal plants for treating cancer is extremely important. Over the past few years, the anti-cancer properties of Nigella Sativa L. have been proven. The aim of the present study was to evaluate, the cytotoxic effect of a nanoemulsion synthesized using N. Sativa L. tincture, against a cancerous cell line as well as its and free radical scavenging activities.
Materials and Methods: The size and zeta potential of the nanoemulsion were determined using particle size analyzer and morphological shape of nano emulsion was visualized by transmission electron microscopy (TEM). The antioxidant activity of nanoemulsions was investigated by the DPPH assay. Cytotoxic effects of the nanoemulsions were assessed by MTT method against A2780 ovarian cancer and umbilical vein endothelial cells (HUVEC) as normal cells. To evaluate the probable molecular mechanism of cell death, acridine orange and propidium iodide staining methods were used for identifying apoptotic cells.
Results: The results obtained from this study showed that the synthesized nanoemulsion had a good and dose-dependent radical scavenging capacity in the DPPH assay (IC50 of about 47μg/ml). Also, the nanoemulsion significantly reduced the bioavailability of A2780 cancerous cells (IC50 of 0.72 μg/ml); however, its toxicity against HUVEC cells was much lower (IC50 > 25 μg/ml). The pro-apoptotic effect of the produced nanoemulsion was confirmed by acridine orange and propidium iodide staining.
Conclusion: Nano emulsions synthesized by N. Sativa L. tincture has a relevant potential antioxidant and anticancer effects and therefore they can be considered and studied as anticancer compounds in future experiments.


1.Amiri MS, Joharchi MR, TaghavizadehYazdi ME. Ethno-medicinal plants used to cure jaundice by traditional healers of Mashhad, Iran. Iran j pharm res. 2014; 13(1): 157.
2.Yazdi MET, Housaindokht MR, Sadeghnia HR, Esmaeilzadeh Bahabadi S, Amiri MS, Darroudi M. Assessment of phytochemical components and antioxidant activity of Rheum turkestanicum Janisch. Studies in Medical Sciences. 2020; 31(2): 75-81.
3.Yazdi ME, Khara J, Husaindokht MR, Reza H, Sadeghnia SE, Amiri MS, Darroudi M Biocomponents and antioxidant activity of Ribes khorasanicum. IJBMS. 2018; 3(3): 99-103.
4.Eisenberg DM, Davis RB, Ettner SL, Appel S, Wilkey S, Van Rompay M, Kessler RC. Trends in alternative medicine use in the United States, 1990-1997: results of a follow-up national survey. Jama. 1998; 280(18): 1569-1575.
5.Yazdi ME, Amiri MS, Hosseini HA, Oskuee RK, Mosawee H, Pakravanan K, Darroudi M. Plant-based synthesis of silver nanoparticles in Handelia trichophylla and their biological activities. Bull Mater Sci. 2019; 42(4): 155.
6.Assi MA, Noor MM, Bachek NF, Ahmad H, Haron AW, Yusoff MS, Rajion MA. The various effects of Nigella Sativa on multiple body systems in human and animals. PJSRR. 2016 ;2(3): 1-9.
7.Darakhshan S, Tahvilian R, Colagar AH, Babolsar I. Nigella sativa: A plant with multiple therapeutic implications. Int J Pharmacognosy. 2015; 2(5): 190-114.
8.Tavakkoli A, Mahdian V, Razavi BM, Hosseinzadeh H. Review on clinical trials of black seed (Nigella sativa) and its active constituent, thymoquinone. J Pharmacopuncture. 2017;20(3):179.
9.Venkatachallam SKT, Pattekhan H, Divakar S, Kadimi US. Chemical composition of Nigella sativa L. seed extracts obtained by supercritical carbon dioxide. Int J Food Sci Technol. 2010; 47(6): 598-605.
10.Tembhurne S, Feroz S, More B, Sakarkar D. A review on therapeutic potential of Nigella sativa (kalonji) seeds. J Med Plant Res. 2014; 8(3): 167-177.
11.Ahmad A, Husain A, Mujeeb M, Khan SA, Najmi AK, Siddique NA, Damanhouri ZA, Anwar F. A review on therapeutic potential of Nigella sativa: A miracle herb. Asian Pac J Trop Biomed. 2013; 3(5): 337-352.
12.Menounos P, Staphylakis K, Gegiou D. The sterols of Nigella sativa seed oil. Phytochemistry. 1986; 25(3): 761-763.
13.Gilani A, Aziz N, Khurram I, Chaudhary K, Iqbal A. Bronchodilator, spasmolytic and calcium antagonist activities of Nigella sativa seeds (Kalonji): a traditional herbal product with multiple medicinal uses. J Pak Med Assoc. 2001; 51(3): 115.
14.Yimer EM, Tuem KB, Karim A, Ur-Rehman N, Anwar F. Nigella sativa L.(black cumin): a promising natural remedy for wide range of illnesses. Evid Based Complement Alternat Med. 2019; 2019.
15.Es-haghi A, Soltani M, Karimi E, Namvar F, Homayouni-Tabrizi M. Evaluation of antioxidant and anticancer properties of zinc oxide nanoparticles synthesized using Aspergillus niger extract. Mater Res Express. 2019; 6(12): 125415.
16.Yazdi ME, Darroudi M, Amiri MS, Hosseini HA, Nourbakhsh F, Mashreghi M, Farjadi M, Kouhi SM, Mousavi SH. Anticancer, antimicrobial, and dye degradation activity of biosynthesised silver nanoparticle using Artemisia kopetdaghensis. Micro nano lett. 2020.
17.Singh Y, Meher JG, Raval K, Khan FA, Chaurasia M, Jain NK, Chourasia MK. Nanoemulsion. Concepts, development and applications in drug delivery. J Control Release 2017; 252: 28-49.
18.Kale SN, Deore SL. Emulsion micro emulsion and nano emulsion: a review. Syst Rev Pharm. 2017; 8(1): 39.
19.Bromley PJ, inventor; Virun Inc, assignee. Pre-spray emulsions and powders containing non-polar compounds. United States patent US 10,016,363. 2018 Jul 10.
20.Baghani M, Es-haghi A. The Antioxidant Activity and Cytotoxic Effects of Amaranthus cruentus-Biosynthesized Silver Nanoparticles Toward MCF-7 Breast Cancer Cell Line. IJBSM. 2019; 19: 20.
21.Es-haghi A, Javadi F, Yazdi MET, Amiri MS. The Expression of Antioxidant Genes and Cytotoxicity of Biosynthesized Cerium Oxide Nanoparticles Against Hepatic Carcinoma Cell Line. Avicenna J Med Biochem. 2019; 7(1): 16-20.
22.Zarei M, Karimi E, Oskoueian E, Es-Haghi A, Yazdi MET. Comparative Study on the Biological Effects of Sodium Citrate-Based and Apigenin-Based Synthesized Silver Nanoparticles. Nutr. Cancer. 2020: 1-9.
23.Mahato R. Nanoemulsion as targeted drug delivery system for cancer therapeutics. J Pharm Sci Pharmacol. 2017; 3(2): 83-97.
24.Kamble SS, Gacche RN. Evaluation of anti-breast cancer, anti-angiogenic and antioxidant properties of selected medicinal plants. Eur J Integr Med. 2019; 25: 13-19.
25.Yazdi ME, Amiri MS, Akbari S, Sharifalhoseini M, Nourbakhsh F, Mashreghi M, Abbasi MR, Modarres M, Es-haghi A. Green Synthesis of Silver Nanoparticles Using Helichrysum graveolens for Biomedical Applications and Wastewater Treatment. BioNanoScience. 2020: 1-7.
26.Darroudi M, Yazdi MET, Amiri MS. Plant-Mediated Biosynthesis of Nanoparticles. 21st Century Nanoscience–A Handbook: CRC Press; 2020. p. 1---18.
27.Bilia AR, Eterno F, Bergonzi MC, Mazzi G, Vincieri FF. Evaluation of the content and stability of the constituents of mother tinctures and tinctures: The case of Crataegus oxyacantha L. and Hieracium pilosella L. J Pharm Biomed Anal. 2007; 44(1): 70-78.
28.Ieri F, Martini S, Innocenti M, Mulinacci N. Phenolic Distribution in Liquid Preparations of Vaccinium myrtillus L. and Vaccinium vitis idaea L. Phytochem Anal. 2013; 24(5): 467-475.
29.Osanloo M, Amani A, Sereshti H, Abai MR, Esmaeili F, Sedaghat MM. Preparation and optimization nanoemulsion of Tarragon (Artemisia dracunculus) essential oil as effective herbal larvicide against Anopheles stephensi. Ind Crops Prod. 2017; 109: 214-9.
30.Sirivibulkovit K, Nouanthavong S, Sameenoi Y. based DPPH assay for antioxidant activity analysis. Anal Sci. 2018; 34(7): 795-800.
31.Van Meerloo J, Kaspers GJ, Cloos J. Cell sensitivity assays: the MTT assay. Cancer cell culture: Springer; 2011. p. 237-45.
32.Baghani M, Es-haghi A. Characterization of silver nanoparticles biosynthesized using Amaranthus cruentus. Bioinspir biomim nan. 2019: 1-8.
33.Salopek B, Krasic D, Filipovic S. Measurement and application of zeta-potential. Rud Geolosko Naft Zb. 1992; 4(1): 147.
34.Nerio LS, Olivero-Verbel J, Stashenko E. Repellent activity of essential oils: a review. Bioresour Technol. 2010; 101(1): 372-378.
35.Shamasi Z, Es-haghi A, Taghavizadeh Yazdi ME, Amiri MS, Homayouni-Tabrizi M. Role of Rubia tinctorum in the synthesis of zinc oxide nanoparticles and apoptosis induction in breast cancer cell line. Nanomed J 2020:-.
36.Shakeel F, Shafiq S, Haq N, Alanazi FK, Alsarra IA. Nanoemulsions as potential vehicles for transdermal and dermal delivery of hydrophobic compounds: an overview. Expert opin drug del. 2012 ;9(8): 953-974.
37.Harwansh RK, Deshmukh R, Rahman MA. Nanoemulsion: Promising nanocarrier system for delivery of herbal bioactives. J Drug Deliv Sci Technol. 2019; 51: 224-233.
38.Viswanathan P, Muralidaran Y, Ragavan G. Challenges in oral drug delivery: a nano-based strategy to overcome. Nanostructures for oral medicine. 2017. p. 173-201.
39.Korak T, Ergül E, Sazci A. Nigella sativa and Cancer: A Review Focusing on Breast Cancer, Inhibition of Metastasis and Enhancement of Natural Killer Cell Cytotoxicity. Curr Pharm Biotechnol. 2020.
40.Majdalawieh AF, Fayyad MW, Nasrallah GK. Anti-cancer properties and mechanisms of action of thymoquinone, the major active ingredient of Nigella sativa. Crit Rev Food Sci Nutr. 2017; 57(18): 3911-3928.
41.Koroleva M, Nagovitsina T, Yurtov E. Nanoemulsions stabilized by non-ionic surfactants: stability and degradation mechanisms. Phys Chem Chem Phys. 2018; 20(15): 10369-10377.
42.Ndisang JF, Vannacci A, Rastogi S. Oxidative stress and inflammation in obesity, diabetes, hypertension, and related cardiometabolic complications. Hindawi; 2014.
43.Rodic S, Vincent MD. Reactive oxygen species (ROS) are a key determinant of cancer’s metabolic phenotype. Int J Cancer. 2018; 142(3): 440-448.
44.Ziech D, Franco R, Pappa A, Panayiotidis MI. Reactive Oxygen Species (ROS)––Induced genetic and epigenetic alterations in human carcinogenesis. Mutat Res 2011; 711(1-2): 167-173.
45.Hileman EO, Liu J, Albitar M, Keating MJ, Huang P. Intrinsic oxidative stress in cancer cells: a biochemical basis for therapeutic selectivity. Cancer Chemother. Pharmacol. 2004; 53(3): 209-19.
46.Rinaldi F, Hanieh PN, Longhi C, Carradori S, Secci D, Zengin G. Neem oil nanoemulsions: characterisation and antioxidant activity. J Enzyme Inhib Med Chem. 2017; 32(1): 1265-1273.
47.Shahbazi Y. Antioxidant, antibacterial, and antifungal properties of nanoemulsion of clove essential oil. J Nanomed Res. 2019; 4(4): 204-208.
48.Eid AM, Elmarzugi NA, Abu Ayyash LM, Sawafta MN, Daana HI. A Review on the Cosmeceutical and External Applications of Nigella sativa. J Trop Med. 2017; 2017.
49.Zaorsky NG, Churilla T, Egleston B, Fisher S, Ridge J, Horwitz E, et al. Causes of death among cancer patients. Ann Oncol. 2017; 28(2): 400-407.
50.Reid BM, Permuth JB, Sellers TA. Epidemiology of ovarian cancer: a review. Cancer Biol Med 2017; 14(1): 9.
51.Jabir NR, Tabrez S, Ashraf GM, Shakil S, Damanhouri GA, Kamal MA. Nanotechnology-based approaches in anticancer research. Int J Nanomedicine. 2012; 7: 4391.
52.Gordaliza M. Natural products as leads to anticancer drugs. Clin Transl Oncol. 2007; 9(12): 767-776.
53.Imran M, Rauf A, Khan IA, Shahbaz M, Qaisrani TB, Fatmawati S, Abu-Izneid T, Imran A, Rahman KU, Gondal TA. Thymoquinone: A novel strategy to combat cancer: A review. Biomed. Pharmacother. 2018; 106: 390-402.
54.Randhawa MA, Alghamdi MS. Anticancer activity of Nigella sativa (black seed)—a review. Am J Chin Med. 2011; 39(06): 1075-1091.
55.Ait Mbarek L, Ait Mouse H, Elabbadi N, Bensalah M, Gamouh A, Aboufatima R. Anti-tumor properties of blackseed (Nigella sativa L.) extracts. Braz J Med Biol Res. 2007; 40(6): 839-847.
56.Hosseini F. Investigating the Effects of Anti-tumor and Anti-inflammatory of Nigella sativa Plant on 4T1 and CT26 Tumor Cell Lines. J Paramed Sci. 2017; 12(3): 0-.
57.Shokrzadeh M, Habibi E, Pourfoladchi N, Mohammadi H. Protective Effects of Trigonella foenum graecum L. Extract Against Cytotoxicity Induced by Cyclophosphamide on Ovarian Cancer (SKOV3) and Normal Cell Lines. J Mazandaran Univ Med Sci. 2017; 27(152): 31-39.
58.Singh G, Marimuthu P, de Heluani CS, Catalan C. Chemical constituents and antimicrobial and antioxidant potentials of essential oil and acetone extract of Nigella sativa seeds. J Sci Food Agric. 2005; 85(13): 2297-2306.