Anticancer activity of silver nanoparticles synthesized using aqueous fruit shell extract of Tamarindus indica on MCF-7 human breast cancer cell line

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Abstract

This present investigation reveals that the Silver Nanoparticles biosynthesizing capability of the fruit shell of pharmacologically important tree Tamarindus indica. An ecofriendly, easy, one step, non-toxic and inexpensive approach is used, where aqueous plant extract acts as a reducing as well as stabilizing agent of Silver Nanoparticles. The Silver Nanoparticles were characterized by UV–Vis spectroscopy, Fourier Transform Infrared Spectroscopy, Energy-Dispersive X-ray, Scanning Electron and Transmission Electron Microscopy and spectroscopy analysis. Surface plasmon resonance of the Nanoparticles was observed at 450 nm in UV–Vis Spectroscopy. Fourier Transform Infrared Spectroscopy result confirms that the plant extract acts as the reducing as well as the capping agent of the Silver Nanoparticles. Transmission Electron Microscopy indicated that the synthesized Nanoparticles are spherical in shape and approximately 20–52 nm in size. Presence of Silver in the Silver Nanoparticles is 36.87% by weight, as confirmed by Energy-Dispersive Spectroscopy (EDS). The synthesized Silver Nanoparticles have cytotoxicity against MCF-7 cell lines. The cytotoxicity study exhibited a dose-dependent effect against breast cancer cells (MCF-7) using MTT assay, the inhibitory concentration (IC50) was found to be 20 μg/ml and their anticancer potential has been discovered using live and dead assay (Ao/EtBr), ROS and Rho123 assay. In conclusion, the results of this study demonstrated that Silver Nanoparticles from Tamarindus indica fruit shell extract may be a potential therapeutic agent for human breast cancer treatment.

Introduction

Nanotechnology is a multidisciplinary science field has the aim to create atomic, molecular and supramolecular materials with enhanced properties. Nanomaterials are used in glucose, DNA and RNA detection, heavy detection, disease diagnosis, disease control and micro-electronics [1]. Various methods are available for preparations of metal nanoparticles are physical, chemical and biological methods. Biological method means synthesis of nanoparticles using bacteria, fungi, algae, yeast and plants [[2], [3], [4]]. The physical and chemical methods have some advantages are that the nanoparticles has narrow size distribution while the main disadvantages are consumption of high energy, time and high cost and usage of toxic chemicals [5]. Biological methods have many advantages over conventional methods like eco-friendly, low cost, lack of downstream processing and high yields. Moreover, the biological methods are otherwise called as green synthesis. Hence, Nanoparticles synthesis through plants extracts offers simple one step reduction process with large scale production. Numerous report on the fabrication of Nanoparticles using plant extracts with significant biological activities [6].

Reduction of metal ions into Nanoparticles using plant extracts is called Phytomining. The exploration of plant extracts for Nanoparticles synthesis is widespread protocol in recent years. Plant extracts are serving as reducing, capping and stabilizing agents. Plant related parts such as leaves, stems, roots, shoots, flowers, barks, seeds and their metabolites have been successfully used for the efficient biosynthesis of Nanoparticles [7]. synthesized Silver Nanoparticles using aqueous extract of Nepetade flersiana plant. Face centered cubic structured Nanoparticles was obtained with the average size 33 nm. This process shows the maximum SPR band at 400 nm. Functional groups carboxylic and alcoholic groups from flavonoids and terpenoids were responsible for the reduction process. In green synthesis of Nanoparticles sunlight was act as a catalyst and garlic extract was used as reducing agent was demonstrated by Rastogi and Arunachalam (2011). Enhanced production of Silver Nanoparticles was achieved by employing Persea Americana seed extract with different concentration. Here different sized Nanoparticles were observed whereas the concentration was increased the size of Silver nanocolloid also increased. While increasing the concentration of extract in Nanoparticles synthesis different colors from yellow to brown was obtained. These different colors are associated with the surface plasmon resonance effect of Silver Nanoparticles. Semi-spherical to large oblongated Nanoparticles with the size 2.3 nm were observed from TEM images [8]. Nanoparticles have potential to cure various diseases and it is an alternative therapeutic agent due to the unique properties based on their morphological characters. In the recent years, Silver Nanoparticles recommended to development of new anticancer drugs due to broad-spectrum of biological activities [[9], [10], [11]]. Nanoparticles are highly toxic to cancerous cells than bulk materials. This is a rapidly increasing the possibilities of using Nanoparticles as therapeutic agents in the diagnosis and treatment of cancer cells. And also, Nanoparticles can be designed to carry drugs and controlled release at the targeted site.

Silver Nanoparticles serve as antitumor agents by decreasing progressive development of tumor cells. This might be because of their inhibitory actions in several signaling cascades liable for the development and pathogenesis of cancer. Taken together, this information recommends that Silver Nanoparticles can actuate cytotoxicity on cancer cells and hindering tumor progression without lethality to normal cells [12]. In this present investigation we have used fruit shell extract of Tamarindus indica for the first time to synthesize silver nanoparticles and its application on anticancer activity. The synthesized nanoparticles were characterized using various spectroscopic and microscopic techniques.

Section snippets

Chemicals

Analytical graded Silver Nitrate, DPPH (2,2-Diphenyl-1-Picrylhydrazyl), Ammonium Molybdate, Hydrogen Peroxide, Ascorbic Acid, MTT reagent (3-(4, 5-dimethylthiazol- 2-yl)-2, 5-diphenyltetrazolium bromide), Dimethyl Sulfoxide (DMSO), Acridine Orange and Ethidium Bromide (Ao/EtBr), Phosphate Buffer Solution (PBS), Rhodamine 123, and other chemicals and solvents were obtained from Sigma Aldrich (Bangalore, India) and Himedia Ltd (Mumbai, (India) with highest purity.

Collection of plant material

Tamarindus indica fruit shells

Cell culture

The breast cancer cell line (MCF-7) was purchased in NCCS PUNE. Cells were grown in high glucose media DMEM using FBS (V/V), 1% penicillin/streptomycin (W/V). The cells were grown in 5% CO2 and 95% humidified atmosphere air.

Cell cytotoxicity (MTT) assay

The cell cytotoxicity was achieved by the MTT assay (3-(4, 5-dimethylthiazol- 2-yl)-2, 5-diphenyltetrazolium bromide). The MCF-7 cells are seeded in 96 well plates (1 × 104 cells/well) and allowed to attach overnight. The next day morning, old media was aspirated with a new

Visual observation

Green synthesis of Silver Nanoparticles using tamarind fruit shell extract was initially confirmed by visual observation. The shell extract was exposed to Silver ion solution the Silver Nanoparticles was formed. Initially, the reaction mixture was appeared pale yellow in color and it changed into brown indicates the formation of Silver Nanoparticles (Fig. 1). After 1 h the reaction mixture changed into dark brown and blackish brown indicates that reduction of Silver ion process is completed.

Conclusion

Silver Nanoparticles have emerged as an important class of nanomaterials for a wide range of industrial and medical applications. Developing biocompatible molecule, using nanotechnology, as an anticancer agent is one of the novel approaches in the field of cancer therapy. The Tamarindus indica (Tamarind) fruit shell extract used for the synthesis of Silver Nanoparticles is a simple, low cost, eco-friendly and large scale production. The UV–Vis spectrum shows the characteristics absorption peak

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