Anticancer activity of pure and silver doped copper oxide nanoparticles against A549 Cell line

https://doi.org/10.1016/j.mtcomm.2022.104462Get rights and content

Highlights

  • Pure and Ag-doped CuO Nanoparticles have been synthesized using green synthesis.

  • Moringa oleifera leaf extract as reducing, capping, and stabilizing agents.

  • The anticancer activity of synthesized Nps were tested against A549 cell line.

  • Cytotoxicity effect for 20% Ag-doped CuO Nps showed a decrease in cell viability.

Abstract

In day-to-day life, green synthesis of metal oxide nanoparticles has still now achieved excellent heed in the field of anticancer activity. In this present study, Moringa oleifera leaf extract functions as reducing, capping, and stabilizing agents for the preparation of pure and silver doped copper oxide nanoparticles of various (5%, 10%, 15%, and 20%) concentrations. The synthesized materials were subjected to various characterization techniques. XRD, reveals that nanoparticles are in the monoclinic phase and the mean crystallite size for pure and Ag-doped CuO nanoparticles of various concentrations is found to be in the range of 15.22–3.67 nm. SEM images show that nanoparticles are relieved from particle size enlargement and EDAX analysis confirmed the existence of elements like Cu, O, and Ag. The biomolecules present in the leaf extract that helps for the bio-reduction of CuO nanoparticles were identified by FT-IR spectrometer. From UV–vis absorption spectra, peaks from 210 to 240 nm were observed. Furthermore, it also exhibits that if the concentration of silver doping increases there will be an increase in the energy bandgap. By MTT assay, the powerful cytotoxic activity for the synthesized pure and Ag-doped CuO nanoparticles of different concentrations has been probed against human lung cancer A549 cells. Among that, 20% of Ag-doped CuO nanoparticles showed a decrease in cell viability with an IC50 value of 15 µg/mL. After treatment with green synthesized nanoparticles, the cell death was assessed through three types of assessments which include AO/EB dual staining, Hoechst staining, and DCFH-DA staining assays in the cell line. The results betrayed that through apoptosis induction, the tested samples treated against human lung cancer A549 cells showed inhibition in cell proliferation and enhances morphological, and cytological alterations via an increased level of ROS accumulation in the NSCLC cell line.

Introduction

Cancer is a disease that divides the abnormal cells uncontrollably which are present in the part of the human body. The World Health Organization (WHO) has comprehensively reported that nearly 10 million humankind are dying because of cancer worldwide [1]. There are 5 major groups of cancer they are: Carcinoma, Leukemia, Sarcoma, Lymphoma, and Myeloma cancers. Carcinoma is one of the most frequent types of cancer among various kinds. It is found on the surface of internal organs and glands that are usually covered by the tissue or skin. Some examples of this are breast cancer, colorectal cancer, prostate cancer, and lung cancer. Among them, 18.4% of all deaths occurred from lung cancer [2]. During the time of diagnosis, approximately 70% of lung cases have a progressive disease but after diagnosis, only 15% of patients are alive [3]. In many countries, the reason for cancer is that the rate of mortality increases mainly due to inhabitants, population, age, growth, and lifestyle of people who feel anxiety, smoke is affected by radiation or environmental pollution and inadequate physical activities [4]. There are several techniques to cure lung cancer including, chemotherapy, targeted drug therapy, radiation therapy, immunotherapy, and surgery. These techniques are used just to halt the growth of multiple cells which can increase the lifetime of patients.

At the same time, the usage of chemotherapy drugs has its demerits like severe side effects, pernicious and high-cost effectiveness [5], [6], [7]. To overcome this, nanomaterials are found to be an important one for the finding of present-day cancer drugs and they played a vital role in the treatment of cancer [8], [9], [10]. Nanoparticles are beneficial in the field of bio-medicine especially in cancer therapy due to their unique properties like large surface-to-volume ratio and nanoscale size in the range of 1–100 nm [11]. Moreover, they also provide an increased amount of drug efficacy with the extended-release of drug matter [12]. Typically, cancer cells are targeted by metal nanoparticles [13].

Researchers also have proved that metal nanoparticles like silver [14], gold [15], cerium [16], iron [17], cobalt [18], zinc oxide [19], and titanium dioxide [20] have a good meritorious anticancer activity. Among them, Copper oxide nanoparticles can kill cancer cells and it has been captivated by all the researchers due to their long everlasting span and they are quite cost-effective [21]. In lung epithelial A549 cells, the cell cycle was arrested by the CuO Nps, from which copper ions are released. Then it persuades DNA damage, started to regulate downwards the cell nuclear antigen, and finally, it gets proliferated [22]. Copper Oxide nanoparticles have distinctive properties towards electrical, optical, chemical, thermal, and biological. Moreover, it has enormous applications in various fields like biomedical, industrial, environmental, and pharmaceutical [23], [24]. Of all metals, Silver is one of the most valuable metals with properties like electronic, chemical, and optical [25] in addition, it has many remarkable therapeutic applications such as anti-angiogenesis, anti-cancer, anti-platelet, antimicrobial, and anti-inflammatory activities [26], [27], [28], [29]. Nanoparticles can be synthesized using both physical and chemical methods [30], [31], [32], [33], [34]. To overcome the drawbacks and the usage of chemical reagents which include noxiousness, and health hazardousness [35], [36], the present study was carried out to synthesize pure and silver doped copper oxide nanoparticles of various concentrations in eco-friendly routeway by M. Oleifera leaf extract. This plant belongs to the family of Moringaceae and it is commonly known as the “Drumstick tree” [37]. The secondary metabolites have played a major role in biological properties [38]. M. Oleifera Leaves contain secondary metabolites such as flavonoids, saponins, terpenoids, tannins, and phenols reduces the Cu2+ ions in the precursor, and also biomolecules such as amino acids and proteins act as an efficient reducing, capping, and stabilizing agents [39], [40]. Related to Copper and Copper Oxide nanoparticles mediated from the plant extract there is numerous research on anticancer activity against lung, breast, colon, and blood that has been reported [41]. As mentioned earlier, Copper, silver, and gold nanoparticles are suitable materials for bio-applications. Among these, we were interested to investigate the biocompatibility for pure and while incorporating silver as a dopant into CuO nanoparticles against human A549 lung cancer of various concentrations using M. Oleifera leaf extract and to study the anticancer activity.

Section snippets

Materials

The chemicals, Copper (II) chloride dihydrate (CuCl2.2 H2O), and Silver nitrate (AgNO3) are of analytical grade, purchased from Sigma Aldrich with 99% and all aqueous solutions have been prepared using double distilled water. 2′, 7′ -dichlorofluorescein-diacetate was purchased from Merck. Dimethyl thiazolyl tetrazolium bromide, dimethyl sulfoxide, acridine orange/ethidium bromide, and Hoechst were obtained from Hi-Media. All the chemicals used for cell cultures are cell culture grades.

Preparation of leaf extract

Moringa

X-ray diffraction

XRD spectra for pure and Ag-doped CuO nanoparticles of various concentrations mediated from M. Oleifera leaf extract was shown in Fig. 2. The X-ray diffraction pattern explains that the peaks show good crystalline nature due to the absence of other peaks like Cu(OH)2, and Ag, and it confirmed the presence of CuO nanoparticles. Furthermore, peak intensities were identical with JCPDS card no. (01–089–5899) and it indicated that the planes were in the monoclinic phase. The average crystallite size

Conclusion

Pure and Ag-doped CuO nanoparticles of various concentrations have been synthesized using M. Oleifera leaf extract as reducing, capping, and stabilizing agents. In contrast to conventional methods, the greener approach is facile, affordable, expeditious, and eco-friendly. Characterization techniques like XRD, SEM, EDAX, FTIR, and UV studies have confirmed the formation of synthesized nanoparticles. Besides, the anticancer activity of pure, and Ag-doped CuO nanoparticles of various

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

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