Plant Mediated Synthesis Cu-Ag-Ru Trimetallic Nanoparticles Using Andrographis Paniculata extract: Antibacterial, Anticancer and Photocatalytic Activities

Introduction

Cancer ranks among the most lethal diseases, resulting in deaths prior to the age of 70 across the globe. According to GLOBOCAN, cancer cases are on the rise, leading to over 9.6 million fatalities worldwide, with predictions indicating further increases by 2040¹. It appears that one in nine individuals is likely to develop cancer during their lifetime. High rates of lung cancer are observed in males, while breast cancer predominates in females. In children, cancers such as lymphoid leukemia affect boys (29.2%) and girls (24.2%).. It was estimated that the incidence of cancer cases will rise from 1.46 million (2022) to 1.57 million(2025)². Factors such as rapid urbanization, an older demographic, sedentary and unhealthy lifestyles, fast-food consumption, and air pollution—both indoor and outdoor—seriously impact individuals in middle- to low-income countries like India ³.

The Indian Council of Medical Informatics and Research (NCDIR) and the National Cancer Registry Programme (NCRP) provide crucial statistics on cancer cases. The data indicates that cancer prevalence is higher among males than females ⁴. Numerous synthetic medications are available, but they often come with side effects. Chemotherapy and radiation therapy have been developed as cancer treatment options, though they also carry side effects⁵. The discovery of drugs through biological methods has led to the identification of many treatments derived from various medicinal plants aimed at addressing diseases⁶. Drugs sourced from medicinal plants have shown effectiveness and minimal side effects⁷.

Andrographis paniculata is a traditional Ayurvedic remedy, part of the Andrographis genus in the Acanthaceae family, which consists of 40 species. Commonly known as the King of Bitter (Kalmegh), it is an annual, branched herb utilized for various health issues⁸. The decoction of this plant is utilized for treating ailments such as fever, colic, the common cold, leucorrhea, prenatal and postnatal care, jaundice, gonorrhea, wounds, and snake bites^9,10. This plant is used to treat respiratory infections, sore throats, and chronic diseases. It naturally possesses antimicrobial¹¹, anti-inflammatory¹², antioxidant¹³, antiallergic and hepatoprotective activity, and nephroprotective activity¹⁴. It was found to have high efficacy in anticancer activity against neuroblastoma (IMR-32) and human cell lines¹⁵. Some of the compounds in Andrographis paniculata, such as 14-deoxyandrographolide and hydroandrographolide, . These compounds, at higher concentrations, have highly effective cytotoxicity towards HPBL ^16,17.

Nanomedicine involves the application of knowledge and tools of nanotechnology to diagnose, treat, and prevent disease with minimal side effects ¹⁸. Nanomaterials are attractive due to their innovative methodologies and glorious applications because of their physical, chemical, biological, optical, and electrical properties. Some metals like palladium, silver, gold, ruthenium, and copper nanoparticles were synthesized by various routes, and the formation of nanoparticles depends upon the particle size and shapes ¹⁹.

Nanomaterials have a wide spectrum of applications in areas ranging from catalytics, photonics, molecular computing, energy storage, fuel cells, tuneable resonant devices, sensors, etc. This is because of increased reactivity in nanoscale materials when compared to their nanosized counterparts²⁰. In fact, it has special properties such as small dimensions, electrical, mechanical, optical, magnetic, and thermal stabilizing properties, and a high surface-to-volume ratio. The surface coating of nanoparticles (NPs) also plays a significant role. Metallic NPs are catagorised into monometallic, dimetallic and polymetallic nanoparticles depending upon the number of metals and oxidation state of the metals. These polymetallic nanoparticles(NPS) have attracted attention because of their enhanced catalytic properties²⁰. The bioengineered NPs are designed to remove harmful particles from the polluted water. Since it is toxic to heavy metals from industries and wastewater from households²¹.

Multi-metallic NPs are generally considered novel materials because they incorporate two or more metals to make alloys. The Multi metallic NPs can be modified to enhance their physicochemical and morphological properties throuigh controlling its structure,temperature,kinetic factor, and shapes to achieve maximum potential and applications. It was concluded that the Multi metallic NPs have specific characteristics that result from the combined effects of individual metals²². The cytotoxicity of metallic NPs synthesized in an aqueous medium of plant extract depends upon the time, dose, etc., particularly their size and shape. It was reported that the smaller size of the NPs was more potent than other shapes. The metallic NPs were closely associated with the generation of reactive oxygen species (ROS). During the DPPH scavenging activity, the diluted NPs were suspended in an aqueous solution of 0.02% to assess the radical scavenging activity²³.

Nanoparticles as drug carrier

Several improvements have been made to chemotherapy over the past 25 years. Some drugs are proven to have excellent efficacy but cause severe side effects and also affect the quality of life. More importantly, these drugs are very expensive. For example, dEPoB was synthesized based on the mechanism of action of paclitaxel and reported to be more efficient than paclitaxel. The discovery of new drugs is a slow process to reduce the existing side effects. However, the ideal chemotherapeutics can be achieved, which requires a high level of concentration of drugs able to concentrate affected cells without disturbing non-targeted tissues and organs. They should be designed in such a way to cause very minimal or no side effects. This concept, also known as ‘ magic bullet’, comes from the experience of German chemist Paul Ehrlich. The magic bullet consists of two components. One should be designed to recognize the target cells and bind to them. Another should play a therapeutic role in this target. In many cases, anticancer payloads, namely radionucleotides, toxins, and chemotherapeutic agents, have been delivered to tumors via monoclonal antibodies²⁴. The various pharmaceutical carriers, including polymers, microcapsules, microparticles, cells, cell ghosts, lipoproteins, liposomes, and micelles, have been used as carriers to multiply the number of drug molecules per target without affecting the normal surrounding cells and tissues. There has been recent progress in the development of nanoparticles. The different kinds of nanoparticles, such as organic, inorganic, and hybrid nanoparticles, are designed to improve the interaction between the biomolecules, controlled transport, and releases²⁵.

Synthesis of Nanoparticles

The various physical and chemical methods, such as electrochemical, ultrasonication, chemical vapor deposition, etc., are significant; the formation of NPs depends upon the process, such as the kinetics of interaction between the metal ions with the reagent, adsorption, influence in morphology, and physiochemical parameters ^26,27. Figure 1. shows the methods of synthesis of nanoparticles.

Figure 1: Different methods of synthesis of nanoparticles. Click here to View Figure

The metallic nanoparticles Ru, Ag, and Pd were produced from the aqueous extract of garlic tunicate leaves. The average dimensions of these TNPs ranged from 50 to 90 nm²⁸. Copper oxide nanoparticles were synthesized using the leaf extract of the Ficus religiosa. Additionally, the anticancer properties can be evaluated at varying concentrations of CuO NPs, which were identified to be 200 µg/mL in A549 cells after 36 hours²⁹. Silver nanoparticles were generated using Andrographis paniculata through an extracellular approach³⁰. Likewise, silver nanoparticles have been produced utilizing numerous plant extracts, including Areca catechu, Maesa calophylla, Maesa laxiflora, Adinandra poilanei, and others. Notably, the six extracts exhibited greater 2,2-diphenyl-1-picryl hydroxyl radical scavenging activity and reducing potential compared to any other extracts. The Ag NPs displayed significant negative zeta potential values, with hydrodynamic sizes ranging from 12.5±1.0 nm to 21.3±4.9 nm. Furthermore, they demonstrated substantial cytotoxic effects on A549 and HeLa cells³¹. Certain metal nanoparticles, including cadmium oxide (CdO), neodymium oxide (Nd2O3), and nickel nanoparticles, were synthesized using extracts of Andrographis paniculata, and these nanoparticles showed effectiveness in anticancer research employing MCF-7 cells^32,33.

Materials and Methods

Materials

Copper sulfate, silver nitrate, and ruthenium chloride were brought from the Sisco Chemical Laboratory with high purity.

Preparation of  TNPS

Approximately 10 grams of fresh Andrographis paniculata leaves were gathered, thoroughly rinsed with tap water to eliminate any contaminants, and then allowed to dry at room temperature. About 250 milliliters of double-distilled or deionized water was used. Around 150 milliliters of this water was added and boiled for one hour at 60 °C to create a plant extract. The resulting solution took on a straw yellow hue. The solution was left overnight and then filtered using Whatman 40 filter paper.

Approximately 3 milliliters of 0.1 N copper sulfate, silver nitrate, and ruthenium chloride were incorporated and heated to 70 °C for one hour. Subsequently, the precursor solution was combined with the extract solution, resulting in a color change from straw yellow to black. The solution was placed on a magnetic stirrer and left still at room temperature. The precipitate that formed was filtered, rinsed with ethanol several times to remove any impurities, and then dried in an oven at 150 °C. Consequently, TNPs were produced and are available for characterization for further analysis.

Figure 2: Preparation of biosynthesized CuO/Ag/Ru TNPs via the leaves extract of A.paniculata. Click here to View Figure

Characterization of TNPs

The biological preparation was initially validated through UV-visible spectroscopy. It was subsequently analyzed using a SHIMADZU-UV 1800 UV-visible spectrophotometer and quartz, utilizing a 10 mm path length. This device is employed to acquire qualitative information about substances via their characteristic absorption patterns present in the samples. The morphology of the nanoparticles (NPs) can be verified using scanning electron microscopy (SEM) and atomic force microscopy (AFM). The SEM (model: JEOL-JSM-IT200) operates at an accelerating voltage range of 0.5 kV to 30 kV, with images captured at various magnifications, including 1500X, 5000X, and 10,000X, among others. The interactions between metals and the biochemical compounds found in the NPs can be explored with Fourier-transform infrared spectroscopy (FTIR). The elemental makeup and ratio of individual metals within the TNPs can be assessed using energy dispersive X-ray spectroscopy (EDX). Additionally, the crystallinity and average size of the NPs can be gauged through X-ray diffraction (XRD). X-ray photoelectron spectroscopy (XPS) (model: K alpha) can be employed to validate the formation of NPs and the oxidation states of the metals in the materials. The decomposition, melting points, purity, and energy transitions may be examined with a thermal analyzer (model: NETZSCH-STA449F3 Jupiter). The temperature can be varied from room temperature up to 1400 °C at a rate of 10,200 °C/min in a volume of 10 ml liquid within a blank atmosphere.

Result And Discussion

UV-Visible Spectroscopy

The UV-visible spectroscopy used to assess the formation of nanoparticles and the absorption of the biological TNPs was measured in the range of 200–400 nm. The strong surface plasmon peaks observed at 210nm and 330nm correspond to metals such as CuO, Ag, and Ru NPs. These resonance peaks confirm the formation of the inner and outer cores of TNPs. Moreover, the formation of NPs can be visually confirmed by the color changes from straw yellow to black, and Figure 3A. represents the UV-visible spectra of TNPs.

Figure 3A: UV-Visible spectra of CuO/Ag/ Ru TNPs Click here to View Figure

FTIR

Different functional groups, including ketones, aldehydes, alkaloids, and terpenoids, in the plant extract contribute to the reduction of metals into metallic nanoparticles (NPs). This research aimed to identify the functional groups present in the NPs, as well as to assess the interaction between the metal and the plant extract. The observed bands at 3360, 2900, 1720, and 1160 cm^-1 correspond to the stretching vibrations of OH, C-H, C=O, C=C, and C-O, respectively. The functional groups found in the TNPs suggest the presence of bioorganic compounds such as phenols, amino acids, and carboxylic acids, among others^33,34. A band detected at 3390 and 1640 cm^-1 is associated with C=O stretching vibrations from carboxyl groups and C=N bending from amide groups. The band around 1590 cm^-1 aligns with the N-H band of primary amines found in the Andrographis paniculata extract. The approximately 2900 cm^–1 band corresponds to phenolic and alcoholic compounds in the Andrographis paniculata extract. The slight overlapping bands in this spectrum indicate that the extract of Andrographis paniculata is integrated with the metal NPs. Additionally, the bands observed below 600 cm^-1 are primarily attributed to the metal-metal interactions within the TNPs³³.

Figure 3B: FTIR spectra of CuO/Ag/ Ru TNPs Click here to View Figure

SEM and EDX

The dimensions and configuration of the TNPs can be assessed through SEM analysis, while EDX provides insights into the elemental composition of each TNP, as depicted in Fig. 3C. Figure 3. C illustrates the SEM analysis at various magnification levels of 1 m, 5 m, 10 m, and 500 m, showing that the shape of the TNPs is spherical, manifesting an agglomerated structure where individual NPs cluster together. In the EDX analysis, the metal NPs are composed of carbon (38.9%), oxygen (43.38%), copper (3.2%), ruthenium (4.7%), and silver (10.21%), respectively.

Figure 3C: shows SEM analysis of TNPs at 10µm,5µm,1µm and 500µm respectively and shows EDX spectra of TNPs. Click here to View Figure

XRD

The material’s lattice, crystallinity, average size, and crystal orientation of the NPs can be investigated using XRD analysis. The observed diffraction sharp peaks positions at 2θ= 32.59⁰, 35.61⁰, 38.78⁰, 48.82⁰, 53.37, 66.31, 68.15, 72.4 were assigned to (110), (-111), (111),(-202),(020),(202),(-113),(-311),(220),(-220), and (311) are consistent with the previous literature (JCPDS No. 01-080-0076) of copper oxide NPs with a monoclinic phase[34]. The prominent diffraction peaks at 2θ = 37.99, 46.29, 64.34, and 77.19 correspond to cubic crystalline Ag NPs with JCPDS No. 04-0783[35]. The diffraction peaks of RU NPs were assigned to hexagonal The average size of the TNPs can be calculated by means of the Debye Scherrer formula, D= Kλ/ Cosθ  where k is the Scherrer constant, λ = 1.5418A, β = 0.06, and cosθ = 0.96[36]. In this formula, (0.90 x 1.5418) / (0.06 x 0.97) = 22.2nm. Thus, the diameter of the crystallite is about 22nm.

Figure 3D: XRD spectra of CuO/Ag/ Ru TNPs. Click here to View Figure

TEM and DLS

From the TEM analysis, it was observed that CuO/Ag/Ru trimetallic nanoparticles composed of spherical, flake, and cup-shaped morphology and the size of the nanoparticles was 25-100 nm(fig.3.E) and in good agreement with the DLS, XRD, and AFM values. The agglomerated form of the nanoparticles was similar to the SEM morphology. Further, these morphologies were compared with the reported previous literature of individual metal nanoparticles. DLS spectra also observed that the maximum intensity was between 1-100 nm with a diameter of 192.5 nm and a standard deviation of 176. Dynamic Light Scattering (DLS) analysis was performed using a Micromeritics instrument, specifically the Nanoplus model. This equipment is capable of measuring the particle sizes of liquid-suspended samples through a dilution method, covering a size range from 0.1 nm to 12.3 µm, with sample concentrations varying from 0.00001% to 40% and a sensitivity for molecular weights as low as 250 Da. DLS provides insights into the dimensions of the nanoparticles produced. Dynamic light scattering illustrates the behavior of nanoparticles as well as their tendency for agglomeration or settling. A light source is introduced into the cell, and the scattered light is captured and analyzed to determine the particle size. The particle size is recorded at an average diameter of 192.5 nm, with a standard deviation of 176 and a refractive index of 1.3 as shown in fig3.F.

Figure 3E: TEM images of CuO/Ag/Ru TNPS. Click here to View Figure

Figure 3F: DLS spectra of CuO/Ag/Ru TNPs. Click here to View Figure

 

AFM

It is an advanced technique to assess the size, shape, and dispersion of TNPs. Here, the sample is dissolved in a suitable solvent and dried as a thin layer on a mica-based glass slide³⁷. Figure 3.G. shows two-dimensional, three-dimensional, and particle-size histograms of TNPs. From the SEM and AFM studies, it was confirmed that the TNPs are spherical-shaped and agglomerated, with an average particle size of about 22 nm.

Figure 3G: AFM spectra of CuO/Ag/ Ru TNPS. Click here to View Figure

XPS

XPS  is used to determine the conformation and oxidation states of metals in TNPs. The binding energy of elements sch as O,Cu, Ag, Ru are given in the table1. The binding energy of  Ru(3d) is 285ev,O(1s) is 532ev,Ag(3d) is 368ev,and  Cu(2p) is 935ev. The XPS spectra of individual elements are given in fig.3.H. The XPS survey spectra the Ag existed as Ag(3d) and Ag(3p³) with the binding energies 368ev and 560ev respectively separated by 200ev.

Table 1: XPS spectra of CuO/Ag/Ru TNPs.

Name	Peak BE	FWHM ev	Area (P)CPS.eV	Atomic %
Ru 3d	285.9	2.4	104601.28	1.32
O 1s	532.45	4.18	1232073.04	95.44
Ag3d	368.41	2.73	355510.88	2.98
Cu2p	935.26	2.22	256830.11	3.23

 

Figure 3H: XPS spectra of Cu(2p), O(1s),Ag(3d),C(1s), Ru(3d) and XPS survey spectra of CuO/Ag/Ru TNPs. Click here to View Figure

Thermal studies (DTA/TG)

DTA/TG is utilized to evaluate the thermal properties of the sample, which include decomposition, melting point, glass transition, oxidation rate, crystallinity degree, purity, transition energy, etc. A sample of 50–100 mg of CuO/Ag/Ru TNPs was placed in an Al₂O₃ crucible, and the temperature was maintained between 20 and 700 °C. As the temperature rises, the material begins to decompose at approximately 300 °C, with the reaction being exothermic, as shown in Fig. 3.i.

Figure 3I: Thermal studies (DTA/TG) of CuO/Ag/Ru TNPs. Click here to View Figure

Photocatalytic Activity of TNPs

The photocatalytic activity of TNPs can be performed using methylene blue dye in a UV-irradiated photoreactor, and this methylene blue (MB) shows characteristic absorption peaks from 200 to 400nm. The decolorization process occurs when UV light is irradiated on the solution to the concentration of X10^-3 M at various concentrations. The effectiveness of degradation process  on the nature of the substance used , time, and the binding of catalyst with the molecules of MB dye that have degraded. This increases the active sites of the photocatalyst surface and free hydroxyl radicals. During this process, the dark blue of the dye diminishes over time because the TNPs act as scavengers that are applied to the active sites of the photocatalyst. Importantly,the formation of reactive oxygen species (ROS) results from the electron-hole pair surface transmission process. These generated ROS are responsible for the photocatalytic activity of TNPs³⁸.

Figure 4: Photocatalytic activity of CuO/Ag/Ru TNPs using Victoria blue dye. Click here to View Figure

Antibacterial Activity of TNPs

The microbial strains of Escherichia coli, Streptococcus pyogenes, Staphylococcus aureus, and Klebsiella pneumoniae used to examine the antibacterial activity of the TNPs at different concentrations are shown in table 2, and the image were given in fig5.A. The zone of inhibition values for TNPs against bacterials strains at three different concentrations (25µl,50µl& 50µl) are graphically represented in the fig 5.B. The mechanism behind the antimicrobial activity is that the higher biological content of the TNPs, indicated by potential disruption of the membrane, damage to DNA, and production of reactive oxygen species such as peroxide, superoxide, inactive species, etc., can eliminate or reduce bacterial growth and, intricately with the membrane, lipids, and enzymes, induce cellular death. The hybrid TNPs exhibited higher antimicrobial activity than the individual metal NPs²⁸.

Table 2: Antibacterial activity of CuO/Ag/Ru TNPs using different pathogens at various concentrations.

Bacterials pathogens	Zoon of inhibition (mm)
	25 μl	50 μl	75 μl	Norfloxacin
Escherichia coli	10±0.10	14±0.13	18±0.14	30±1.12
Streptococcus pyogenes	19±0.18	20±0.17	25±0.17	29±1.13
Staphylococcus aureus	10±0.12	13±0.12	15±0.15	30±1.10
Klebsiella pneumoniae	17±0.16	20±0.17	22±0.17	32±1.14

 

Figure 5A: The graph of maximum antibacterial activity of Cu/Ag/Ru TNPs against E. coli, S. pyogenes, S. aureus, and K. pneumonia. Click here to View Figure

Figure 5B: Antibacterial activity of CuO/Ag/Ru TNPs using  strains of E. coli, S. pyogenes, S. aureus, and K. pneumonia. Click here to View Figure

Anticancer Activity

The yellow compound 3-4,5-dimethylthiozol-2-Yl-2,5-diphenyltetrazoliumbromide (MTT) is transformed by mitochondrial dehydrogenase in healthy cells, producing a quantifiable purple product. Living cells possess NAD(P) H-dependent reductase, which converts the MTT reagent into formazan, resulting in a deep purple color. The crystals of  Formazan crystals are dissolved with the solubilizing solution, and the absorbance are usually recorded between 500 and 600 nm by a plate reader. A total content was dissolved in a  50 mg of MTT with approximately 10 mL of PBS. After vortexing for one minute, it was filtered through 0.45-micron filters. The flask was wrapped in aluminum foil to protect it from light, given that MTT is sensitive to light. This solution was stored about 4 °C for further incvestigations.

For the cell viability assessment, PA-1 viable cells were collected and counted with a hemacytometer, then diluted in DMEM medium to achieve a cell density of 1 × 10⁴ cells/ml. They were placed in 96-well plates, with one well per cell, and incubated for 24 hours to facilitate attachment. Subsequently, PA-1 cells were treated with a control and various concentrations (10–70 µg/ml) of aqueous extracts were administered to each well. The cells were subjected to inubation at 37°C in a wet condition where it containing 95% air and 5% CO₂ for 24 hours. Following the incubation period, the drug-treated cells were thoroughly washed with fresh culture medium, and MTT (5 mg/ml in PBS) dye was added to each well. This content was subjected incubation for  4 hours at 37°C.

The purple precipitate of formazan that formed was dissolved in 100 µl of concentrated DMSO, and cell viability was assessed by measuring absorbance at 540 nm using a multi-well plate reader. The results were expressed in terms of percentage of values viable cells compared to the control values. The half-maximum inhibitory concentration (IC₅₀) values were determined, and the optimal doses were evaluated at various time points^39,40.

The IC₅₀ values were derived from the dose-response curve of the aqueous extract, where a 50% reduction in cytotoxicity was compared to that of control cells. All experiments were conducted at least three times, with three replicates each. The MTT assay results at varying concentrations are presented in Table 3, and the graphical representation of the MTT assay for CuO/Ag/Ru TNPs tested against MCF-7 cells is illustrated in Figure 6.A. The images of both the treated and control groups from the MTT assay for CuO/Ag/Ru TNPs against MCF-7 cells are displayed in Figure 6.B.

Inhibitory of cell proliferation %

Statistical analysis

 The statistical comparisons were performed by one-way analysis of variance (ANOVA), followed by Duncan’s multiple range test (DMRT), using SPSS version 12.0 for Windows (SPSS Inc., Chicago; http://www.spss.com). The values are considered statistically significant if the p value is less than 0.05^41,42.

 Table 3: MTT assay of CuO/Ag/Ru TNPs  at different concentrations.

MCF-7
Ni	Blank	0	100	200	300	400	500	600	700	800
A	0.06	1.380	1.532	1.481	1.270	1.0261	0.837	0.6048	0.687	0.632
B	0.08	1.670	1.477	1.204	1.260	1.2494	0.978	0.8291	0.823	0.765
C	0.03	1.7526	1.432	1.1871	1.133	1.0924	1.2089	1.2089	0.734	0.654
		1.601	1.480	1.291	1.221	1.123	0.947	0.881	0.748	0.684

Figure 6A: Graphical representation of MTT assay of CuO/Ag/Ru TNPs against MCF-7cells. Click here to View Figure

Figure 6B: The images of treated and controlled MCF-7 breast cancer cells using CuO/Ag/Ru TNPs. Click here to View Figure

Antioxidant Activities

DPPH (free radical scavenging activity), the DPPH molecules has stable nitrogen radicals used to study the free radical quenching activity of different antioxidants. The DPPH assay was performed with the reference substance ascorbic acid acts as positive control. The TNPs were dissolved with suitable solvent like deionised water at different concentrations as shown in table4. The antioxidant activity of TNPs increases with increasing concentrations. The measurement was performed at five different concentrations and plotted to calculate R² value as shown in fig 8&9. The antioxidant activity is the ability to protect from oxidative damages and IC₅₀ values were found to be 107.8µg/ml and R² value is 0.912. Furthermore, this result is good agreement with the antioxidant activity values of individual nanoparticles. The percentage of radical scavenging activity (%) can be calculated using the following equations.

Radical scavenging activity (%) = OD_control -OD_sample    X 100

Table 4: Antioxidant activity of TNPs at different concentrations

Test	Concentration of Sample (mg/ml)	% of Inhibition of Nanoparticles	Ascorbic acid
DPPH assay	50	31.3	40
	100	39.2	50
	500	48.8	60
	200	59.2	70
	250	68.6	80
IC50VALUE		107.8	100

 

Figure 7: Graph of antioxidant activity of TNPS with positive control Click here to View Figure

Figure 8: Graphical representation of antioxidant activity of CuO/Ag/Ru TNPs  at different concentrations. Click here to View Figure

Conclusion

This green method provide great opportunities to develop more complex nanomaterials with good yield with less minimal of reactants. The trimetallic nanoparticles are usually greater performance in properties with enourmous applications and outcomes. This nanoparticles shows highly selective in catalytic action in photocatalytic activities. It has unique antibacterial activity against bacterial strains. It exhibit superior anticancer activities against human breast cancer cells even in minimum concentrations. This nanoparticles has distinct shape can help bind with the active site of the drug molecules. It can be used as potential drug carrier with minimum harmness to the normal cells.

Acknowledgment

I thanful to my Research supervisor , Dr.S.Srinivasan, Associate Professor, Annamalai University .

Funding Sources

The author(s) received no financial support for the research, authorship, and/or publication of this article.

Conflict of Interest

The author(s) do not have any conflict of interest.

References

1. Bray F, Ferley J, Soarjomataram I , Siegel LR, Torre AL , Jemal A“ Global cancer statistics 2018 : GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries”, CA Cancer J.Clin. 2018;68:394-424.
   CrossRef
2. Krishnan Sathishkumar, Meesha Chaturvedi, Priyanka Das, S.Stephen and Prashant Mathur,  “Cancer incidence estimates for 2022 &  projection for 2025; Result from National cancer registry programme India”, Indian J Med Res 2022, 156, 598-607.
   CrossRef
3. Vaitheeswaran Kulothungan, Krishnan Sathishkumar, sravya Leburu, Thilagavathi Ramamoorthy , Santhappan Stephen, Dharmapa Basavajappa , Nifty Tomy, Rohith Mohan, Geetha R. Menon, and Prashant Mathur “Burden of cancers in India – estimates of cancer crude incidence, YLLS, YLDS and DALYs for 2021 and 2025 based on National cancer registry program, 2022,22: 527“
   CrossRef
4. Cherian Varghese “ Cancer prevention and control in India “
5. Hogland HC, Haemotological complications of cancer chemotherapy . Semki Oncol. 1982; 9: 95-102.
6. .Joselin J, Jeeva S, Andrographis paniculata : A review of its  traditional uses, phytochemistry and pharmacology . Med Aromat plants 3:169.
7.  Fransworth NR : Screening plants for new medicines , chapter 9, edited by Wilson E, Washington D.C, National Academy press; 1988.
8. Perry L M, Medicinal plants of  East and South east Asia : attributed properties and uses. Cambridge : MIT Press ; 1980.
9. Agbonlahor Okhuarolea , Joyco Ehrizoghi Faloda Osayemwenre Erharuyi , Vincent Imieji , Abiodun Falodun, Peter Langer, Harnessing the medicinal properties of Andrographis paniculata for disease and beyond : a review of its phytochemistry and pharmacology , Asian Pae J Trop Dis 2014; 4(3): 213-222.
   CrossRef
10. Zaiden .MR, Noor , Rain A, Badrul. AR, Adlin A, Norazah. A,and Zakiah.I, In vitro screening of five local medicinal plants for antibacterial activity using disc diffusion method, Trop Biomed, 2005; 22: 165-170.
11.  Abu – Ghefreh,. AA , Canatan. H, and Ezeamuzie CI, In vivo and invitro anti-inflammatory effects of Andrographolide , Int Immunopharmacol, 2009;9:313-318. 
    CrossRef
12. Trinedi NP, and  Rawal.UM, H, Hepatoprotective and antioxidant properties of Andrographis paniculata (Nees) in BHC- induced liver damage in mice, Indian J Exp Biol, 2001;39: 41-46
13. Vetriselvan.S, Rajamanickam .V,Muthappan M, and Gnanasekaran D et al,. Hepatoprotective effects of aqueous extract of Andrographics paniculata against CCl₄ induced hepatotoxicity in allerno wistar rats; Asian Journal of Pharmaceutical and clinical Research, 2011; 4(3);93-94.
14. Rajeshkumar Shanmugam, Mohammedi brahim Ponnanikajamideen, M. Nagalingam, Mahendran Vanaja , Anticancer activity of Andrographis paniculata leaves extract against microblastime (IMF-32) and human colon (HT-29) cancer cell line , world journal of pharmacy and pharmaceutical sciences , 205.
15. Ajaya Kumar, K. Sridevi, N.Vijaya Kumar, S.Nanduri, S. Rajagopal, Anticancer and immunostimulatory compounds from Andrographis paniculata , Journal of of ethnopharmacology 92(2004) 291-295.
    CrossRef
16.  MM. Rahman, F. Islam, S.Afsana Mim et al., Multifunctional therapeutic approach of nanomedicines against inflammation in cancer and ageing , Journal of nanomaterials,2022, vol.2022, Article ID 4217529, 19 pages.
    CrossRef
17.  A.M. Abu-Dief, L.H.Abdel Rahman. MA, Abd-El Sayed, MM. Zikry and A.Nafady,Green synthesis of Ag nanoparticles utilizing Delonix Regia extract as anticancer antimicrobial agents, chemistry select ,2021, pp.13263-13266.
    CrossRef
18. C.N.Rao, H.S.S.Ramakrishna Matte, Rakesh Yoggu and A.Govindaraj , Recent progress in the synthesis of inorganic nanoparticles , Dalton Trans, 2012,41,5089.
    CrossRef
19. Sharma G, Kumar.A, Sharma. S, Nushand.M, Prakash Dwaivedi.R, Alothman ,Z.A, Mola, G.T, Novel development of nanoparticles to bimetallic nanoparticles and their composites : A review J. King Savd Univ.Sci. 2019, 31, 259-269.
    CrossRef
20. Sana Abbas, Saima Nasreen , Adeela Haron, Muhammad Aqeel Ashraf, Synthesis of Silver and Copper Nanoparticles from plants and applications as Adsorbents for Naphthalene decontamination , Saudi Journal of Biological sciences 27(2020)1016-1023.
    CrossRef
21. Nagaraj Basavagowda and Kwang- Hyun Baek , Multimetallic nanoparticles as Alternative Antimicrobial agents ; challenges and perspectives molecules. 2021 ,26,912,
    CrossRef
22. Eun -Young Ahn, Younie Park, Anticancer prospects of silver nanoparticles green- synthesized by plant extracts, Material science and Engineering C116(2020) 111 253, 2020.
    CrossRef
23. Kohler G and Milstein C:  Continuous cultures of fuel cells secreting antibody of predefined specificity . Nature 75(suppl.I) : 381-394,1975.
24.  Langer R : Drug delivery and targeting . Nature 392 : 5-10, 1998.
    CrossRef
25.  Jin Zhang, Christopher Q. Lens, Michael Post, Benott Simard , Yues Deslandes, and Tsung Han Hsieh , Design of Nanoparticles as drug carriers for cancer therapy, cancer genomics & proteomics : 147-158, 2006.
26. D.R.Bhumkar, H.M.Joshi, Sastry,V.B., Pokharkar, chitosan seduced gold nanoparticles as novel carriers for transmucosal delivery of insulin, Pharm. Res. 24(8)(2007)1415-1426.
    CrossRef
27. Prasad Govindrao Jamkharde , Namerata W.Ghule, Abdul Haque Baner, Mohan G. Kalaskar,Metal nanoparticle synthesis : An overview on methods of preparation, advantage and disadvantages and applications, Journal of Drug Delivery Science and Technology 53(2019), 101174. 
    CrossRef
28. Shaimaa Hussein, Ayman M.Mahmond, Hassan A. Elegebaly, Omnia Magdyn Hendawy, Emad H.M.Hassaneim, Shaima M.N. Moustafa , Nasssar F.Alotaibi, and Amr M.Nassar. Journal of chemistry, volume 2022, 14 pages.3
    CrossRef
29. Renu Sankar, Ramasamy Maheswari, Selvaraju Karthik, Kanchi Subramanian Shivashangari, Vilwanathan Ravikumar, Anticancer activity of Ficus religiosa engineered copper oxide nanoparticles , Materials science and Engineering C44(2014) 234-239, 2014.
    CrossRef
30. C.Pannerselvam, S.Ponarulselvam and K.Murugan , Potential Anti-plasmodial Activity of Synthesized Silver Nanoparticles using Andrographics paniculata Nees (Acanthaceae), Archives of Applied Science Research, 2011, 3(6) : 208-217.
31. M.Sundrarajan , V.Muthulakshmi , Green synthesis of ionic liquid mediated Neodymium oxide nanoparticles by Andrographis paniculata leaves extract for effective biomedical applications. S7213-3437(20)31065-4.
32.  K.Karthik, S.Dhanuskodi, C.Gobinath, S.Prabukumar, S.Sivaramakrishnan, Andrographis paniculata extract mediated green synthesis of CdO nanoparticles and its electrochemical and antibacterial studies. J.Mater Sci : Mater Electron (2017)28: 7991-8001.
    CrossRef
33. Zahra Vaseghi, Omid Tavakoli, Ali Nematollahzadeh, Rapid
    biosynthesis of novel Cu/Cr/Ni trimetallic oxide nanoparticles with antimicrobial activity : (2018) 1898-1911.
    CrossRef
34. Hojat Veisi, Bikash Karmakar, Taiebeh Tamoradi, Rerza Tayebee, Sami Jadifar, Shahram Lotfi, Behrooz Maleki & Saba Hemmati, Bio- inspired synthesis of palladium nanoparticles fabricated magnetic Fe3O4 nanocomposite over Fritillaria imperialis flower extract as an efficient recyclable catalyst for the reduction of nitroarenes, Scientific reports, 4515, 2020.
    CrossRef
35. Awadesh Kumar Mishra, Kavindra Nath Tiwari, Rajesh Saini, Pradeep Kumar, Sunil Kumar Mishra , Virendra Bahadur Yadav, Gopath Nath, Journal of inorganic and organomateallic polymers and materials (2020)30: 2266-2278.
    CrossRef
36. Haritha Meruvu, Meena Vangalapati, Seema Chaitanya Chippada and Srinivasa Rao Bammidi, Synthesis and characterization of Zinc oxide nanoparticles and its antimicrobial acti vity against Bacillus substilis and Escherichia coli , Rasayan J.Chem, Vol.4,No.1.2017, 217-222.
37. Ram Prasad and Vyshnava Satyanarayana Swamy , Antibacterial Activity of Silver Nanoparticles Synthesized by Bark Extract of Syguim cumini , Journal of Nanoparticles, Volume 2013, 6 pages.
    CrossRef
38. Absulmohsen Ali Alsehri and Maqsood Ahmad Malik, Facile one pot Biogenic synthesis of Cu-Co-Ni Trimetallic Nanoparticles for enhanced photocatalytic Dye Degradation , 2020.
    CrossRef
39. J.Jayachitra, M.Akshaya, N.Pushpa, R.Suja Pandian and C. Ruth Christiya,Anticancer activity, UV-Visible and FTIR analysis of Herbal formulation. Annals of R.S.C.B, ISSN: 1583-6258, Vol.25, Issue 6, 2021, pages 5636-5655.
    CrossRef
40. Tim Mosmann, Rapid Colorimetric Assay for Cellular Growth and Survival ; Application to Proliferation and Cytotoxicity Assays, Journal of immunological Methods, 65(1983), 55-63.
    CrossRef
41. Yinmom Htun, Shinji Nakamura, Yasuhiro Nakao, Tsutomu Mitsuie, Kenichi Ohta, Makota Arioka, Takayuki Yokota, Eri Inove, Toi Tsuchiya Kosuke Koyano, Yukihiko Konishi, Takanori Miki, Masaki Veno and Takashi Kusaka, Conflicting findings on the effectiveness of hydrogen therapy for ameliorating vascular leakage in a 5-day hypoxic- isochemic survival piglet model , Scientific report, 2023, 13: 10486.
    CrossRef
42. Peter Larsson, Hama Engquist, Jana Biermann, Elisabeth Werener Ronnerman, Eva Forssell- Aronsson, Amiko Kovaes, Per Karlson, Khalil Helon, and Toshima Z.Parris, Optimization of cell viability assays to improve replicability and reproducibility of cancer drugs sensitivity screens, Scientific reports, 2020, 5798. 
    CrossRef

---

This work is licensed under a Creative Commons Attribution 4.0 International License.