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In-vitro antibacterial, antioxidant and cytotoxic potential of gold nanoparticles synthesized using novel Elaeocarpus ganitrus seeds extract

In the present study, we have follwed the hydrothermal path for the synthesis of gold nanoparticles (Au NPs) from the biomaterial Elaeocarpus ganitrus seeds extract, which is a rapid, eco-friendly, non-chemical way. The prepared NPs were thoroughly analysed by powder x-ray diffraction and high resolution transmission electron microscopy studies and were also tested for anticancer studies. Journal of Science: Advanced Materials and Devices 6 (2021) 127e133 Contents lists available at ScienceDirec

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  1. Journal of Science: Advanced Materials and Devices 6 (2021) 127e133 Contents lists available at ScienceDirect Journal of Science: Advanced Materials and Devices journal homepage: www.elsevier.com/locate/jsamd Original Article In-vitro antibacterial, antioxidant and cytotoxic potential of gold nanoparticles synthesized using novel Elaeocarpus ganitrus seeds extract S.P. Vinay a, *, Udayabhanu b, H.N. Sumedha b, M. Shashank b, G. Nagaraju b, N. Chandrasekhar a a Research and Development Center, Department of Chemistry, Shridevi Institute of Engineering and Technology, Tumakuru 572106, Karnataka, India b Energy Materials Research Laboratory, Department of Chemistry, Siddaganga Institute of Technology, Tumakuru 572103, Karnataka, India a r t i c l e i n f o a b s t r a c t Article history: In the present study, we have follwed the hydrothermal path for the synthesis of gold nanoparticles (Au Received 1 April 2020 NPs) from the biomaterial Elaeocarpus ganitrus seeds extract, which is a rapid, eco-friendly, non-chemical Received in revised form way. The prepared NPs were thoroughly analysed by powder x-ray diffraction and high resolution 5 September 2020 transmission electron microscopy studies and were also tested for anticancer studies. Besides, the Accepted 8 September 2020 antioxidant, antibacterial and anticancer properties of Au NPs were studied. In vitro studies revealed the Available online 22 September 2020 dose-dependent cytotoxic effect of Au NPs. The prepared nanoparticles showed good cytotoxic impact against a prostate cancer (PC-3) cells line. The evidences of the current study lead to the synthesis of Keywords: Elaeocarpu sganitrus novel and cost-effective drugs from Elaeocarpus ganitrus seeds extract by using the bio approach. Hydrothermal synthesis © 2020 The Authors. Publishing services by Elsevier B.V. on behalf of Vietnam National University, Hanoi. Gold nanoparticles This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). Prostate cancer Cancer nanomedicine 1. Introduction looked into techniques to grow new successful antimicrobial op- erators that beat protections of these microorganisms and that are The research on nanotechnology is an auspicious area for likewise cost effective [4]. Subsequently, selective medications, cancerous therapy and diagnostics in the biomedical approach. which are less lethal, eco-accommodating and cheap, should be Cancer is a dangerous and threatening disease in the world and investigated. The present work deals with Au NPs that exhibit does increases the morbidity and mortality of human life. Malig- effective cytotoxicity against prostate cancer (PC-3) cells. The level nancy treatment incorporates chemotherapy, radiotherapy and of cell was essentially diminished by expanding the concentration medical procedure and is an incredibly factor in its introduction, from 20 to 100 mg/mL. improvement and result, since now and again these treatments A tree, called Elaeocarpus ganitrus, is an enormous evergreen fizzle or cause the reoccurrence of tumor cells [1]. Likewise, greater expansively leaved tree whose seeds are customarily utilized for parts of the disease drugs kill both harmful and ordinary cells in an petition globules in Hinduism. The seeds are called Rudraksha, or irregular way [2]. Many degenerative diseases of aging such as Rudraksh [5]. E. ganitrus seeds are usually found in the foothills of brain malfunction, cataracts, cardiovascular diseases and cancer, icy Himalayan Mountains in South-East Asia, the Australian part of are generated by the over production of free radicals inside the New Guinea, Indonesia, Nepal, Hawaii, and Guam. E. ganitrus trees body which is caused by oxidative stress. The free radicals and are rarely found in South India [6]. Even if found, they usually do receptive oxygen species are to be deactivated before they harm not yield E. ganitrus seeds. However, two E. ganitrus trees in Udupi cells. Further, by the unexpected increment in the bacterial Kakkunje Garodi yield E. ganitrus seeds during the whole year. The obstruction against numerous anti-toxins [3], researchers have seeds are secured by an external husk of blue shading when they are completely ready, and, therefore, are otherwise called blueberry dots [7,8]. * Corresponding author. E-mail address: s.pvinay143@gmail.com (S.P. Vinay). Peer review under responsibility of Vietnam National University, Hanoi. https://doi.org/10.1016/j.jsamd.2020.09.008 2468-2179/© 2020 The Authors. Publishing services by Elsevier B.V. on behalf of Vietnam National University, Hanoi. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
  2. S.P. Vinay, Udayabhanu, H.N. Sumedha et al. Journal of Science: Advanced Materials and Devices 6 (2021) 127e133 Phytochemical constituents present in E. ganitrus are beaker and then the solution mixture is allowed to stir for 30 min isoelaeocarpine, rudrakine, elaeocarpidine, epiisoelaeocarpiline, to get a uniform solution. Later this mixture is transferred into a alkaloids, flavonoids, carbohydrates, tannins, quercetin, proteins, 50 mL Teflon lined vessel and was fitted in a stainless steel ethanol, ellagic acid, fat and gallic acid [9,10]. Au is a significant and autoclave and maintained at 180 ± 5 C for 3 hr. After completion a kind inorganic substance. and in this light it is impossible to miss of the reaction, the autoclave was allowed to cool to RT naturally. it since it attributes to novel applications in different fields of sci- After centrifugation, the settled nanoparticles were washed with ence and innovation [11]. Moreover, piezoelectric, pyroelectric, ethanol and water for several times. Further, the synthesized Au optoelectronics, catalysis and semiconducting properties are very NPs were dried and put away for further experiments [31]. important features of Au NPs [12]. Due to this, Au is a symbolic material which is used in the field of light-emitting diodes, bio- 2.2. Characterization sensors, diluted and ferromagnetic materials for spintronic solar cells, photo-catalysis, transistors and which also acts as an anti- X-ray diffraction (XRD) data were recorded in the Rigaku Smart oxidant and anti-bacterial agent [13]. Lab. Transmission Electron Microscopy (TEM) (JEOL 3010) was used Nanoparticles can be synthesised by various methods like to detect the size and shape of the nanoparticles. combustion [14e21], the biosynthesis route [22], microwave irra- diation [23e25], hydrothermal synthesis [26] andultrasonic irra- 3. Results and discussion diation [27,28]. Herein, we report the synthesis of gold nanoparticles using biomaterial Elaeocarpus ganitrus seeds extract 3.1. XRD study under the hydrothermal synthesis method. In this reaction capping and reducing agent is Elaeocarpus ganitrus seeds extract by Fig. 1a represents the X-ray diffraction measurement (XRD) of reducing the Chloroauric acid (HAuCl4) to the Au NPs. The charac- Au NPs which gives information about geometric and internal terisation was done by using the XRD and TEM analysis and, later, atomic arrangements. The cubic nature is determined by the the synthesised material was tested for Antibacterial, Antioxidant characteristic planes of crystalline NPs. As depicted in this figure, and Anticancer activity against the Human Prostate Cancer (PC-3) there are four main characteristic diffraction peaks at 2q¼38 , cell line. Table 1 2. Experimental Rietveld refined structural parameters for Au NPs. Au NPs Au (JCPDS No.: 65e2870) All chemicals used were of analytical grade. The Chloroauric acid Crystal system Cubic (HAuCl4) was purchased from E-Merck India Ltd. E. ganitrus seeds Space group Pm-3m (221) were collected from Bramhabyidarekal Garodi Kakkunje, Udupi, Lattice parameters (A ) Karnataka, India. The E. ganitrus seed material was shade dried and a¼b¼c 4.0272 powdered into 100 mesh size and stored at ambient temperature in a¼b¼l 90O an airtight container. Unit cell volume (A 3) 67.365 Atomic coordinates x 0.0000 2.1. Preparation of aunanoparticles y 0.0000 z 0.0000 The preparation of gold nanoparticles using Elaeocarpus Refinement parameters RP 43.3 ganitrus seeds extract proceeded under the hydrothermal syn- RWP 45.7 thesis method [29]. The Au NPs was synthesized by Elaeocarpus RExp 32.3 ganitrus seeds extract [30]. The precursor used was Chloroauric c2 2.0 acid (HAuCl4) and Elaeocarpus ganitrus seeds extract was used as S 1.4136 RBragg 0.984 a reducing agent for synthesis. In a typical hydrothermal syn- RF 0.459 thesis, 33 mL Elaeocarpus ganitrus seeds extract was taken in a QD 1.7382 beaker with constant stirring. Add 330 mg of HAuCl4 into the Fig. 1. (a). PXRD pattern of the Au NPs. (b). Rietveld refined pattern. 128
  3. S.P. Vinay, Udayabhanu, H.N. Sumedha et al. Journal of Science: Advanced Materials and Devices 6 (2021) 127e133 Fig. 2. (a), (b) TEM images, (c) HRTEM image, (d) Histogram show the range of Particle size distribution. 44 , 64 , and 77 that confirm that Au NPs with lattice planes functional positions and occupancy, are summarized in Table 1. (111), (200), (220) and (311) are face centered cubes (FCC). The structure of the examined data is well matched with the Debye-Scherrer’s equation is used to find out the average crys- Joint committee on Powder Diffraction Standards (JCPDS) card tallite size which was found to be 18 nm [32]. Fig. 1b illustrates number 65-2870 [33]. The high pure and intense peaks in the the Rietveld refinement examination to find out the lattice XRD plot show the formation of Au NPs without any other parameter of the Au NPs. The refined parameters for Au NPs, like impurities. Fig. 3. Bar diagram of inhibition zone of antibacterial activity of Au NPs. 129
  4. S.P. Vinay, Udayabhanu, H.N. Sumedha et al. Journal of Science: Advanced Materials and Devices 6 (2021) 127e133 Table 2 Antibacterial activity of Au NPs against pathogenic bacteria. S. No Treatment Pseudomonas desmolyticum Staphylococcus aureus (mean ± SE) (mean ± SE) 1 Control NA NA 2 Au NPs (1000 mg/mL) 9.47 ± 0.30 11.50 ± 0.29 3 Au NPs (1500 mg/mL) 15.50 ± 0.30 18.50 ± 0.29 4 Ciprofloxacin (5 mg/mL) 19.63 ± 0.17 22.17 ± 0.44 Values are the mean ± SE of inhibition zone in mm. NA Symbols represent no antibacterial activity was found in this work. 3.2. Transmission electron microscopy (TEM) studies Fig. 2 shows the TEM analysis of Au NPs in optimal conditions. The nanoscale and uniform distribution of Au NPs were examined by the different images of the TEM analysis. The shape of the obtained NPs is quasi-spherical and the mean particle size is found to be 30.34 ± 0.56 nm. The Au materials d-spacing value is obtained by the HR-TEM images which give clear information about the d-spacing value (0.14 nm), which value belongs to the (111) plane [34]. 4. Antibacterial activity The synthesized Au nanoparticles show a good level of in- hibition which was studied for two different bacterial strains: Gram-negative (P.desmolyticum) and Gram-positive (S.aureus). A significant antibacterial efficiency against bacteria is found, as Fig. 4. Percentage inhibition of DPPH radical. shown in Fig. 3. Table 2 shows the different sizes of zone Fig. 5. Shows the cytotoxicity of Au NPs against PC-3 cancer cell line (a ¼ Untreated, b ¼ Treated with standard and c ¼ Treated with nanoparticles). Fig. 6. (a). Percentage of viability in PC-3 cell line with different concentrations of Au NPs. (b). Percentage of cell inhibition in PC-3 cell line. 130
  5. S.P. Vinay, Udayabhanu, H.N. Sumedha et al. Journal of Science: Advanced Materials and Devices 6 (2021) 127e133 Scheme 1. Schematic mechanism of anticancer activity for Au NPs. inhibition of pathogens. The plate results show the good zone to control the DPPH free radical scavenging activities with IC50 of inhibition which proves the improved anti-bacterial activity. value of 372 mg/ mL (Fig. 4). 5. Antioxidant activity 6. Anticancer activity DPPH, having the characteristic absorption at 517 to 520 nm, is The current work is in concurrence with the ideal application for an antioxidant with a stable free radical which is stable at room cancer treatment which suppresses or prevents carcinogenic pro- temperature. Using this radical, the scavenging activity of Au NPs gression. Figs. 5 and 6 show the anticancer activity in terms of was studied. The extent of radical scavenging was absorbed by the the cell viability against the PC-3 cell line. To estimate the effect of decrease in the absorption, which determines the percentage of Au NPs concentration (20-100 mg/mL) on the cell viability of the PC- scavenging of free radicals or inhibition. The Au NPs has the ability 3 cell line, the MTT assay was performed. As the concentration of Au Table 3 Cytotoxicity of Hydrothermal synthesis of Au NPs against PC-3 Cancer cell line. Cell line Standard Concentration of Au NPs sample (mg/mL) IC50 Cisplatin (mg/mL) Control 20 40 60 80 100 PC-3 5.14 ± 0.32 100 92.83 ± 0.33 80.00 ± 0.29 53.50 ± 0.29 31.83 ± 0.17 3.33 ± 0.17 64.23 ± 0.33 131
  6. S.P. Vinay, Udayabhanu, H.N. Sumedha et al. Journal of Science: Advanced Materials and Devices 6 (2021) 127e133 NPs increased the percentage of cell death in PC-3 cells gradually [14] S.P. Vinay, Udayabhanu, G. Nagaraju, C.P. Chandrappa, N. Chandrashekar, Novel Gomutra (Cow urine) mediated synthesis of silver oxide nanoparticles increases which is represented in Scheme 1 [35,36]. Different and their enhanced Photocatalytic, Photoluminescence and antibacterial concentrations of 20, 40, 60, 80 and 100 mg/mL of Au NPs were studies, J. Sci.: Adv. Mater. Dev. 4 (2019) 392e399, https://doi.org/10.1016/ exposed to PC-3 cells. To evaluate the IC50 values, Curve software j.jsamd.2019.08.004. has been used which is represented in Table 3 [37]. The obtained [15] R. Chauhan, A.S. Kumar, R. Chaudhary, Structure and optical properties of Zn1- xNixO nanoparticles by coprecipitation method, J. Optoelectron. Biomed. evidences clearly show that the Au NPs appreciably control the Mater. 3 (2011) 17e23. cancer cells with a moderate concentration. As the concentration of [16] S.P. Vinay, N. Chandrasekhar, C.P. Chandrappa, Biosynthesis of silver nano- Au NPs against the PC-3 cell line increases the percentage of live particles using leaves extracts of Acalypha hispida burm.f. and study of their antibacterial activity, Int. Res. J. Nat. Appl. Sci. 4 (7) (2017) 9e19. cells was reduced [38e40]. [17] S.P. Vinay, N. Chandrasekhar, C.P. Chandrappa, Eco-friendly approach for the green synthesis of silver nanoparticles using flower extracts of sphagneticola 7. Conclusion trilobata and study of antibacterial activity, Int. J. Pharm. Biol. Sci. (IJPBS) 7 (2) (2017) 145e152. [18] S.P. Vinay, N. Chandrasekhar, C.P. Chandrappa, One-step green synthesis of This study has led to the preparation of Au NPs using the extract silver nanoparticles using flower extract of Tabebuia argentea Bur. & K. 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