Abstract
The unique properties of zinc oxide nanoparticles (ZnO-NPs) produced using plant extract make them attractive for use in medical as well as industrial applications. In this study, ZnO-NPs were synthesized using Hertia intermedia extract as the reducing and stabilizing agent followed by characterization and evaluation of its biological potency. Field emission scanning electron microscopy (FESEM) image showed spherical nanoparticles with a size range of 20–80 nm. UV-vis spectroscopy displayed absorption peaks at 362.67 nm which is one of the characteristic features of ZnO-NPs. FT-IR spectra confirmed the presence of some phytoconstituents as capping agents to stabilize the nanoparticles. MTT assay showed cytotoxicity of ZnO-NPs against Caco-2 (IC50 177 μg/mL), SH-SY5Y (IC50 184 μg/mL), MDA-MB-231 (IC50 168 μg/Ml, and HEK-293 (IC50 240 μg/mL) cell lines. Using 7-dichlorodihydrouorescein diacetate (DCFH-DA) assay, significant production of reactive oxygen species (ROS) was measured after 24 h of treatment with 200 μg/mL ZnO-NPs that is indicative of ZnO-NP-mediated oxidative stress. Induction of apoptosis/necrosis in ZnO-NPs-treated cells was determined using annexin V-PE/7-AAD staining. Furthermore, expression analysis of pro-apoptotic gene Bax and anti-apoptotic gene Bcl-2 by real-time PCR showed 10-fold increase in expression of Bax and 16-fold decrease in expression of Bcl-2 after exposure of cells to ZnO-NPs. Well diffusion method did not show effective antibacterial activities of synthesized ZnO-NPs against both gram-negative and gram-positive bacteria. All the results confirm that the ZnO-NPs synthesized in the present work are a potential candidate to induce ROS and oxidative stress that lead to cytotoxicity in cell lines.
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Muhammad, W., Khan, M. A., Nazir, M., Siddiquah, A., Mushtaq, S., Hashmi, S. S., & Abbasi, B. H. (2019). Papaver somniferum L. mediated novel bioinspired lead oxide (PbO) and iron oxide (Fe2O3) nanoparticles: in-vitro biological applications, biocompatibility and their potential towards HepG2 cell line. Materials Science and Engineering: C, 103, 109740.
Mazzola, L. (2003). Commercializing nanotechnology. Nature Biotechnology, 21(10), 1137–1143.
Hoseini, S. J., Darroudi, M., Oskuee, R. K., Gholami, L., & Zak, A. K. (2015). Honey-based synthesis of ZnO nanopowders and their cytotoxicity effects. Advanced Powder Technology, 26(3), 991–996.
Hadia, N., García-Granda, S., & García, J. R. (2014). Effect of the temperature on structural and optical properties of zinc oxide nanoparticles. Journal of Nanoscience and Nanotechnology, 14(7), 5443–5448.
Padalia, H., & Chanda, S. (2017). Characterization, antifungal and cytotoxic evaluation of green synthesized zinc oxide nanoparticles using Ziziphus nummularia leaf extract. Artificial Cells, Nanomedicine, and Biotechnology, 45(8), 1751–1761.
Vigneshwaran, N., Kumar, S., Kathe, A., Varadarajan, P., & Prasad, V. (2006). Functional finishing of cotton fabrics using zinc oxide–soluble starch nanocomposites. Nanotechnology, 17(20), 5087.
Jain, N., Bhargava, A., Tarafdar, J. C., Singh, S. K., & Panwar, J. (2013). A biomimetic approach towards synthesis of zinc oxide nanoparticles. Applied Microbiology and Biotechnology, 97(2), 859–869.
Tarafdar, J., Agrawal, A., Raliya, R., Kumar, P., Burman, U., & Kaul, R. (2012). ZnO nanoparticles induced synthesis of polysaccharides and phosphatases by Aspergillus fungi. Advanced Science, Engineering and Medicine, 4(4), 324–328.
Prashanth, G., Prashanth, P., Nagabhushana, B., Ananda, S., Krishnaiah, G., Nagendra, H., Sathyananda, H., Rajendra Singh, C., Yogisha, S., & Anand, S. (2018). Comparison of anticancer activity of biocompatible ZnO nanoparticles prepared by solution combustion synthesis using aqueous leaf extracts of Abutilon indicum, Melia azedarach and Indigofera tinctoria as biofuels. Artificial Cells, Nanomedicine, and Biotechnology, 46(5), 968–979.
Sangeetha, G., Rajeshwari, S., & Venckatesh, R. (2011). Green synthesis of zinc oxide nanoparticles by aloe barbadensis miller leaf extract: Structure and optical properties. Materials Research Bulletin, 46(12), 2560–2566.
Alaghemand, A., Khaghani, S., Bihamta, M. R., Gomarian, M., & Ghorbanpour, M. (2018). Green synthesis of zinc oxide nanoparticles using Nigella sativa L. extract: the effect on the height and number of branches. Journal of Nanostructures, 8(1), 82–88.
Saha, R., Subramani, K., Raju, S. A. K. P. M., Rangaraj, S., & Venkatachalam, R. (2018). Psidium guajava leaf extract-mediated synthesis of ZnO nanoparticles under different processing parameters for hydrophobic and antibacterial finishing over cotton fabrics. Progress in Organic Coatings, 124, 80–91.
Luque, P., Nava, O., Soto-Robles, C., Vilchis-Nestor, A., Garrafa-Galvez, H., & Castro-Beltran, A. (2018). Effects of Daucus carota extract used in green synthesis of zinc oxide nanoparticles. Journal of Materials Science: Materials in Electronics, 29(20), 17638–17643.
Ngoepe, N., Mbita, Z., Mathipa, M., Mketo, N., Ntsendwana, B., & Hintsho-Mbita, N. (2018). Biogenic synthesis of ZnO nanoparticles using Monsonia burkeana for use in photocatalytic, antibacterial and anticancer applications. Ceramics International, 44(14), 16999–17006.
Rajeshkumar, S., Kumar, S. V., Ramaiah, A., Agarwal, H., Lakshmi, T., & Roopan, S. M. (2018). Biosynthesis of zinc oxide nanoparticles using Mangifera indica leaves and evaluation of their antioxidant and cytotoxic properties in lung cancer (A549) cells. Enzyme and Microbial Technology, 117, 91–95.
Mahendra, C., Murali, M., Manasa, G., Ponnamma, P., Abhilash, M., Lakshmeesha, T., Satish, A., Amruthesh, K., & Sudarshana, M. (2017). Antibacterial and antimitotic potential of bio-fabricated zinc oxide nanoparticles of Cochlospermum religiosum (L.). Microbial Pathogenesis, 110, 620–629.
Umar, H., Kavaz, D., & Rizaner, N. (2019). Biosynthesis of zinc oxide nanoparticles using Albizia lebbeck stem bark, and evaluation of its antimicrobial, antioxidant, and cytotoxic activities on human breast cancer cell lines. International Journal of Nanomedicine, 14, 87.
Sukri, S. N. A. M., Shameli, K., Wong, M. M.-T., Teow, S.-Y., Chew, J., & Ismail, N. A. (2019). Cytotoxicity and antibacterial activities of plant-mediated synthesized zinc oxide (ZnO) nanoparticles using Punica granatum (pomegranate) fruit peels extract. Journal of Molecular Structure, 1189, 57–65.
Abbasi, B. H., Shah, M., Hashmi, S. S., Nazir, M., Naz, S., Ahmad, W., Khan, I. U., & Hano, C. (2019). Green bio-assisted synthesis, characterization and biological evaluation of biocompatible ZnO NPs synthesized from different tissues of Milk thistle (Silybum marianum). Nanomaterials, 9(8), 1171.
Happy, A., Soumya, M., Kumar, S. V., Rajeshkumar, S., Sheba, R. D., Lakshmi, T., & Nallaswamy, V. D. (2019). Phyto-assisted synthesis of zinc oxide nanoparticles using Cassia alata and its antibacterial activity against Escherichia coli. Biochemistry and Biophysics Reports, 17, 208–211.
Senthilkumar, N., NandhaKumar, E., Priya, P., Soni, D., Vimalan, M., & Potheher, I. V. (2017). Synthesis, anti-bacterial, antiarthritic, anti-oxidant and in-vitro cytotoxicity activities of ZnO nanoparticles using leaf extract of Tectona Grandis (L.). New Journal of Chemistry, 41, 10347–10356.
Sirelkhatim, A., Mahmud, S., Seeni, A., Kaus, N. H. M., Ann, L. C., Bakhori, S. K. M., Hasan, H., & Mohamad, D. (2015). Review on zinc oxide nanoparticles: antibacterial activity and toxicity mechanism. Nano-Micro Letters, 7(3), 219–242.
Doğan SŞ, Kocabaş A (2019) Green synthesis of ZnO nanoparticles with Veronica multifida and their antibiofilm activity. Human & Experimental Toxicology 0960327119888270.
Rauf, M. A., Oves, M., Rehman, F. U., Khan, A. R., & Husain, N. (2019). Bougainvillea flower extract mediated zinc oxide’s nanomaterials for antimicrobial and anticancer activity. Biomedicine & Pharmacotherapy, 116, 108983.
Alam F, Suleman S Study of phytochemistry and antioxidant activity of Hertia intermedia (Boiss.) Flowers of Balochistan.
Esmaeili, S., Hamzeloo-Moghadam, M., Ghaffari, S., & Mosaddegh, M. (2014). Cytotoxic activity screening of some medicinal plants from south of Iran. Research Journal of Pharmacognosy, 1(4), 19–25.
McGaw, L., & Eloff, J. (2005). Screening of 16 poisonous plants for antibacterial, anthelmintic and cytotoxic activity in vitro. South African Journal of Botany, 71(3), 302–306.
Jakupovic, J., Bohlmann, F., & Grenz, M. (1989). Furoeremophilanes from Hertia pallens. Phytochemistry, 28(11), 3231–3232.
Akhgar, M., Ghazanfari, D., & Shariatifar, M. (2012). Chemical composition of the essential oil of Hertia intermedia (Boiss.) O. Kuntze from Iran. Journal of Essential Oil-Bearing Plants, 15(3), 360–363.
Akhgar, M., Shariatifar, M., Akhgar, A., Moradalizadeh, M., & Faghihi-Zarandi, A. (2012). Chemical composition and antibacterial activity of the leaf essential oil from Hertia intermedia. Chemistry of Natural Compounds, 48(2), 329–331.
Soltanian, S., Sheikhbahaei, M., Mansour, M., & Behjat, K. K. (2020). Evaluation of anticancer, antioxidant and antibacterial properties of methanol extract of three Acantholimon Boiss. species. Avicenna Journal of Phytomedicine articles in press. https://doi.org/10.22038/AJP.2020.15805.
Soltanian, S., Sheikhbahaei, M., & Mohamadi, N. (2017). Cytotoxicity evaluation of methanol extracts of some medicinal plants on P19 embryonal carcinoma cells. J Appl Pharm Sci, 7(7), 142–149.
Soltanian, S., Mohamadi, N., Rajaei, P., Khodami, M., & Mohammadi, M. (2019). Phytochemical composition, and cytotoxic, antioxidant, and antibacterial activity of the essential oil and methanol extract of Semenovia suffruticosa. Avicenna Journal of Phytomedicine, 9(2), 143.
Li, H.-B., Jiang, Y., Wong, C.-C., Cheng, K.-W., & Chen, F. (2007). Evaluation of two methods for the extraction of antioxidants from medicinal plants. Analytical and Bioanalytical Chemistry, 388(2), 483–488.
Singh, N., & Haque, F. Z. (2016). Synthesis of zinc oxide nanoparticles with different pH by aqueous solution growth technique. Optik, 127(1), 174–177.
Ravindran, C., Manokari, M., & Shekhawat, M. S. (2016). Biogenic production of zinc oxide nanoparticles from aqueous extracts of Duranta erecta L. World Scientific News, 28, 30–40.
Safawo, T., Sandeep, B., Pola, S., & Tadesse, A. (2018). Synthesis and characterization of zinc oxide nanoparticles using tuber extract of anchote (Coccinia abyssinica (lam.) Cong.) for antimicrobial and antioxidant activity assessment. OpenNano, 3, 56–63.
Bulatov, E., Sayarova, R., Mingaleeva, R., Miftakhova, R., Gomzikova, M., Ignatyev, Y., Petukhov, A., Davidovich, P., Rizvanov, A., & Barlev, N. A. (2018). Isatin-Schiff base-copper (II) complex induces cell death in p53-positive tumors. Cell Death Discovery, 4(1), 1–9.
Marimoutou, M., Le Sage, F., Smadja, J., d’Hellencourt, C. L., Gonthier, M.-P., & Robert-Da Silva, C. (2015). Antioxidant polyphenol-rich extracts from the medicinal plants Antirhea borbonica, Doratoxylon apetalum and Gouania mauritiana protect 3T3-L1 preadipocytes against H 2 O 2, TNFα and LPS inflammatory mediators by regulating the expression of superoxide dismutase and NF-κB genes. Journal of Inflammation, 12(1), 10.
Cui, D., Liang, T., Sun, L., Meng, L., Yang, C., Wang, L., Liang, T., & Li, Q. (2018). Green synthesis of selenium nanoparticles with extract of hawthorn fruit induced HepG2 cells apoptosis. Pharmaceutical Biology, 56(1), 528–534.
Yuvakkumar, R., Suresh, J., Saravanakumar, B., Nathanael, A. J., Hong, S. I., & Rajendran, V. (2015). Rambutan peels promoted biomimetic synthesis of bioinspired zinc oxide nanochains for biomedical applications. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 137, 250–258.
Ogunyemi, S. O., Abdallah, Y., Zhang, M., Fouad, H., Hong, X., Ibrahim, E., Masum, M. M. I., Hossain, A., Mo, J., & Li, B. (2019). Green synthesis of zinc oxide nanoparticles using different plant extracts and their antibacterial activity against Xanthomonas oryzae pv. Oryzae. Artificial Cells, Nanomedicine, and Biotechnology, 47(1), 341–352.
Van Engeland, M., Nieland, L. J., Ramaekers, F. C., Schutte, B., & Reutelingsperger, C. P. (1998). Annexin V-affinity assay: a review on an apoptosis detection system based on phosphatidylserine exposure. Cytometry: The Journal of the International Society for Analytical Cytology, 31(1), 1–9.
Zimmermann, M., & Meyer, N. (2011). Annexin V/7-AAD staining in keratinocytes (pp. 57–63). In: Mammalian Cell Viability. Springer.
Makarov, V., Love, A., Sinitsyna, O., Makarova, S., Yaminsky, I., Taliansky, M., & Kalinina, N. (2014). “Green” nanotechnologies: synthesis of metal nanoparticles using plants. Acta Naturae (англоязычная версия), 6(1 (20)), 35–44.
Kumari, R., Barsainya, M., & Singh, D. P. (2017). Biogenic synthesis of silver nanoparticle by using secondary metabolites from Pseudomonas aeruginosa DM1 and its anti-algal effect on Chlorella vulgaris and Chlorella pyrenoidosa. Environmental Science and Pollution Research, 24(5), 4645–4654.
Chouhan, H. S., & Sharma, A. (2014). International journal of green and herbal chemistry. International Journal, 3(2), 425–433.
Gnanasangeetha, D., & Thambavani, D. S. (2013). Biogenic production of zinc oxide nanoparticles using Acalypha indica. Journal of Chemical, Biological and Physical Sciences (JCBPS), 4(1), 238.
Manokari, M., & Shekhawat, M. S. (2015). Biogenesis of zinc oxide nanoparticles using aqueous extracts of Hemidesmus indicus (l.) R. Br. Int J Res Stud Microbiol Biotechnol, 1(1), 20–24.
Prashanth, G., Prashanth, P., Bora, U., Gadewar, M., Nagabhushana, B., Ananda, S., Krishnaiah, G., & Sathyananda, H. (2015). In vitro antibacterial and cytotoxicity studies of ZnO nanopowders prepared by combustion assisted facile green synthesis. Karbala International Journal of Modern Science, 1(2), 67–77.
Al-Sheddi, E. S., Farshori, N. N., Al-Oqail, M. M., Al-Massarani, S. M., Saquib, Q., Wahab, R., Musarrat, J., Al-Khedhairy, A. A., & Siddiqui, M. A. (2018). Anticancer potential of green synthesized silver nanoparticles using extract of Nepeta deflersiana against human cervical cancer cells (HeLA). Bioinorganic Chemistry and Applications, 2018, 9390784.
Zhang, G., Gurtu, V., Kain, S. R., & Yan, G. (1997). Early detection of apoptosis using a fluorescent conjugate of annexin V. Biotechniques, 23(3), 525–531.
Mohanta, Y. K., Panda, S. K., Jayabalan, R., Sharma, N., Bastia, A. K., & Mohanta, T. K. (2017). Antimicrobial, antioxidant and cytotoxic activity of silver nanoparticles synthesized by leaf extract of Erythrina suberosa (Roxb.). Frontiers in Molecular Biosciences, 4, 14.
Kajani, A. A., Zarkesh-Esfahani, S. H., Bordbar, A.-K., Khosropour, A. R., Razmjou, A., & Kardi, M. (2016). Anticancer effects of silver nanoparticles encapsulated by Taxus baccata extracts. Journal of Molecular Liquids, 223, 549–556.
Wang, C., Hu, X., Gao, Y., & Ji, Y. (2015). ZnO nanoparticles treatment induces apoptosis by increasing intracellular ROS levels in LTEP-a-2 cells. BioMed Research International, 2015, 423287.
Rosarin, F. S., Arulmozhi, V., Nagarajan, S., & Mirunalini, S. (2013). Antiproliferative effect of silver nanoparticles synthesized using amla on Hep2 cell line. Asian Pacific Journal of Tropical Medicine, 6(1), 1–10.
Bai, D.-P., Zhang, X.-F., Zhang, G.-L., Huang, Y.-F., & Gurunathan, S. (2017). Zinc oxide nanoparticles induce apoptosis and autophagy in human ovarian cancer cells. International Journal of Nanomedicine, 12, 6521.
Saliani, M., Jalal, R., & Goharshadi, E. K. (2016). Mechanism of oxidative stress involved in the toxicity of ZnO nanoparticles against eukaryotic cells. Nanomedicine Journal, 3(1), 1–14.
Manke, A., Wang, L., & Rojanasakul, Y. (2013). Mechanisms of nanoparticle-induced oxidative stress and toxicity. BioMed Research International, 2013, 942916.
Fu, P. P., Xia, Q., Hwang, H.-M., Ray, P. C., & Yu, H. (2014). Mechanisms of nanotoxicity: generation of reactive oxygen species. Journal of Food and Drug Analysis, 22(1), 64–75.
Zare, E., Pourseyedi, S., Khatami, M., & Darezereshki, E. (2017). Simple biosynthesis of zinc oxide nanoparticles using nature's source, and it's in vitro bio-activity. Journal of Molecular Structure, 1146, 96–103.
Ciftci, H., TÜRK, M., TAMER, U., Karahan, S., & Menemen, Y. (2013). Silver nanoparticles: Cytotoxic, apoptotic, and necrotic effects on MCF-7 cells. Turkish Journal of Biology, 37(5), 573–581.
Oltval, Z. N., Milliman, C. L., & Korsmeyer, S. J. (1993). Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programed cell death. Cell, 74(4), 609–619.
Krishnaraj, C., Jagan, E., Ramachandran, R., Abirami, S., Mohan, N., & Kalaichelvan, P. (2012). Effect of biologically synthesized silver nanoparticles on Bacopa monnieri (Linn.) Wettst. Plant growth metabolism. Process Biochemistry, 47(4), 651–658.
Baharara, J., Namvar, F., Ramezani, T., Mousavi, M., & Mohamad, R. (2015). Silver nanoparticles biosynthesized using Achillea biebersteinii flower extract: apoptosis induction in MCF-7 cells via caspase activation and regulation of Bax and Bcl-2 gene expression. Molecules, 20(2), 2693–2706.
Salehi, S., Shandiz, S. A. S., Ghanbar, F., Darvish, M. R., Ardestani, M. S., Mirzaie, A., & Jafari, M. (2016). Phytosynthesis of silver nanoparticles using Artemisia marschalliana Sprengel aerial part extract and assessment of their antioxidant, anticancer, and antibacterial properties. International Journal of Nanomedicine, 11, 1835.
Ahamed, M., Akhtar, M. J., Raja, M., Ahmad, I., Siddiqui, M. K. J., AlSalhi, M. S., & Alrokayan, S. A. (2011). ZnO nanorod-induced apoptosis in human alveolar adenocarcinoma cells via p53, survivin and bax/bcl-2 pathways: Role of oxidative stress. Nanomedicine: Nanotechnology, Biology and Medicine, 7(6), 904–913.
Brayner, R., Ferrari-Iliou, R., Brivois, N., Djediat, S., Benedetti, M. F., & Fiévet, F. (2006). Toxicological impact studies based on Escherichia coli bacteria in ultrafine ZnO nanoparticles colloidal medium. Nano Letters, 6(4), 866–870.
Vivek, R., Thangam, R., Muthuchelian, K., Gunasekaran, P., Kaveri, K., & Kannan, S. (2012). Green biosynthesis of silver nanoparticles from Annona squamosa leaf extract and its in vitro cytotoxic effect on MCF-7 cells. Process Biochemistry, 47(12), 2405–2410.
Khan, M. F., Hameedullah, M., Ansari, A. H., Ahmad, E., Lohani, M., Khan, R. H., Alam, M. M., Khan, W., Husain, F. M., & Ahmad, I. (2014). Flower-shaped ZnO nanoparticles synthesized by a novel approach at near-room temperatures with antibacterial and antifungal properties. International Journal of Nanomedicine, 9, 853.
Shaik, M. R., Khan, M., Kuniyil, M., Al-Warthan, A., Alkhathlan, H. Z., Siddiqui, M. R. H., Shaik, J. P., Ahamed, A., Mahmood, A., & Khan, M. (2018). Plant-extract-assisted green synthesis of silver nanoparticles using Origanum vulgare L. extract and their microbicidal activities. Sustainability, 10(4), 913.
Ahmadi Shadmehri, A., Namvar, F., Miri, H., Yaghmaei, P., & Nakhaei Moghaddam, M. (2019). Assessment of antioxidant and antibacterial activities of zinc oxide nanoparticles, graphene and graphene decorated by zinc oxide nanoparticles. International Journal of Nano Dimension, 10(4), 350–358.
Nagajyothi, P., Sreekanth, T., Tettey, C. O., Jun, Y. I., & Mook, S. H. (2014). Characterization, antibacterial, antioxidant, and cytotoxic activities of ZnO nanoparticles using Coptidis Rhizoma. Bioorganic & Medicinal Chemistry Letters, 24(17), 4298–4303.
Y-g, Y., Wang, Y.-h., Xing, H.-H., & Gurunathan, S. (2017). Quercetin-mediated synthesis of graphene oxide–silver nanoparticle nanocomposites: a suitable alternative nanotherapy for neuroblastoma. International Journal of Nanomedicine, 12, 5819.
Acknowledgments
The authors thank Dr. Mansour Mirtadzadini at Department of Biology, Faculty of Science, Shahid Bahonar University of Kerman, for plant collection and identification.
Funding
This study was funded by Bam University of Medical Science, Bam, Iran (grant number: 97000081).
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Soltanian, S., Sheikhbahaei, M., Mohamadi, N. et al. Biosynthesis of Zinc Oxide Nanoparticles Using Hertia intermedia and Evaluation of its Cytotoxic and Antimicrobial Activities. BioNanoSci. 11, 245–255 (2021). https://doi.org/10.1007/s12668-020-00816-z
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DOI: https://doi.org/10.1007/s12668-020-00816-z