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      Rapid Biosynthesis of Silver Nanoparticles Using Pepino ( Solanum muricatum) Leaf Extract and Their Cytotoxicity on HeLa Cells

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          Abstract

          Within nanotechnology, gold and silver nanostructures have unique physical, chemical, and electronic properties [ 1, 2], which make them suitable for a number of applications. Moreover, biosynthetic methods are considered to be a safer alternative to conventional physicochemical procedures for both the environmental and biomedical applications, due to their eco-friendly nature and the avoidance of toxic chemicals in the synthesis. For this reason, employing bio routes in the synthesis of functionalized silver nanoparticles (FAgNP) have gained importance recently in this field. In the present study, we report the rapid synthesis of FAgNP through the extract of pepino ( Solanum muricatum) leaves and employing microwave oven irradiation. The core-shell globular morphology and characterization of the different shaped and sized FAgNP, with a core of 20–50 nm of diameter is established using the UV-Visible spectroscopy (UV-vis), field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM) and Zeta potential and dynamic light scanning (DLS) studies. Moreover, cytotoxic studies employing HeLa (human cervix carcinoma) cells were undertaken to understand FAgNP interactions with cells. HeLa cells showed significant dose dependent antiproliferative activity in the presence of FAgNP at relatively low concentrations. The calculated IC 50 value was 37.5 µg/mL, similar to others obtained for FAgNPs against HeLa cells.

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          Most cited references 64

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          Unique cellular interaction of silver nanoparticles: size-dependent generation of reactive oxygen species.

          The rapid advancement of nanotechnology has created a vast array of engineered nanomaterials (ENMs) which have unique physical (size, shape, crystallinity, surface charge) and chemical (surface coating, elemental composition and solubility) attributes. These physicochemical properties of ENMs can produce chemical conditions to induce a pro-oxidant environment in the cells, causing an imbalanced cellular energy system dependent on redox potential and thereby leading to adverse biological consequences, ranging from the initiation of inflammatory pathways through to cell death. The present study was designed to evaluate size-dependent cellular interactions of known biologically active silver nanoparticles (NPs, Ag-15 nm, Ag-30 nm, and Ag-55 nm). Alveolar macrophages provide the first defense and were studied for their potential role in initiating oxidative stress. Cell exposure produced morphologically abnormal sizes and adherence characteristics with significant NP uptake at high doses after 24 h. Toxicity evaluations using mitochondrial and cell membrane viability along with reactive oxygen species (ROS) were performed. After 24 h of exposure, viability metrics significantly decreased with increasing dose (10-75 microg/mL) of Ag-15 nm and Ag-30 nm NPs. A more than 10-fold increase of ROS levels in cells exposed to 50 microg/mL Ag-15 nm suggests that the cytotoxicity of Ag-15 nm is likely to be mediated through oxidative stress. In addition, activation of the release of traditional inflammatory mediators were examined by measuring levels of cytokines/chemokines, including tumor necrosis factor (TNF-alpha), macrophage inhibitory protein (MIP-2), and interleukin-6 (IL-6), released into the culture media. After 24 h of exposure to Ag-15 nm nanoparticles, a significant inflammatory response was observed by the release of TNF-alpha, MIP-2, and IL-1beta. However, there was no detectable level of IL-6 upon exposure to silver nanoparticles. In summary, a size-dependent toxicity was produced by silver nanoparticles, and one predominant mechanism of toxicity was found to be largely mediated through oxidative stress.
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            Geranium leaf assisted biosynthesis of silver nanoparticles.

            Development of biologically inspired experimental processes for the synthesis of nanoparticles is evolving into an important branch of nanotechnology. In this paper, we report on the use of Geranium (Pelargonium graveolens) leaf extract in the extracellular synthesis of silver nanoparticles. On treating aqueous silver nitrate solution with geranium leaf extract, rapid reduction of the silver ions is observed leading to the formation of highly stable, crystalline silver nanoparticles in solution. Transmission electron microscopy analysis of the silver particles indicated that they ranged in size from 16 to 40 nm and were assembled in solution into quasilinear superstructures. The rate of reduction of the silver ions by the geranium leaf extract is faster than that observed by us in an earlier study using a fungus, Fusarium oxysporum, thus highlighting the possibility that nanoparticle biosynthesis methodologies will achieve rates of synthesis comparable to those of chemical methods. This study also represents an important advance in the use of plants over microorganisms in the biosynthesis of metal nanoparticles.
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              Rapid biological synthesis of silver nanoparticles using plant leaf extracts.

              Five plant leaf extracts (Pine, Persimmon, Ginkgo, Magnolia and Platanus) were used and compared for their extracellular synthesis of metallic silver nanoparticles. Stable silver nanoparticles were formed by treating aqueous solution of AgNO(3) with the plant leaf extracts as reducing agent of Ag(+) to Ag(0). UV-visible spectroscopy was used to monitor the quantitative formation of silver nanoparticles. Magnolia leaf broth was the best reducing agent in terms of synthesis rate and conversion to silver nanoparticles. Only 11 min was required for more than 90% conversion at the reaction temperature of 95 degrees C using Magnolia leaf broth. The synthesized silver nanoparticles were characterized with inductively coupled plasma spectrometry (ICP), energy dispersive X-ray spectroscopy (EDS), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and particle analyzer. The average particle size ranged from 15 to 500 nm. The particle size could be controlled by changing the reaction temperature, leaf broth concentration and AgNO(3) concentration. This environmentally friendly method of biological silver nanoparticles production provides rates of synthesis faster or comparable to those of chemical methods and can potentially be used in various human contacting areas such as cosmetics, foods and medical applications.
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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                Materials (Basel)
                Materials (Basel)
                materials
                Materials
                MDPI
                1996-1944
                28 April 2016
                May 2016
                : 9
                : 5
                Affiliations
                [1 ]Instituto Interuniversitario de Investigación de Reconocimiento Molecular y Desarrollo Tecnológico (IDM), Unidad Mixta Universitat Politècnica de València-Universitat de València, Camino de Vera s/n, Valencia 46022, Spain; mogormo@ 123456upvnet.upv.es (M.G.); elazgi@ 123456upvnet.upv.es (E.A.); fsanceno@ 123456upvnet.upv.es (F.S.); mmarcos@ 123456qim.upv.es (M.D.M.)
                [2 ]CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Valencia 46022, Spain
                [3 ]Materials Chemistry Laboratory, Department of Chemistry, Gulbarga University, Gulbarga, Karnataka 585106, India; kujalliravi@ 123456gmail.com (R.B.); raman.dms@ 123456gmail.com (A.V.)
                [4 ]Biological Research Innovation Centre and Solutions LLP, Bengaluru, Karnataka 56004, India
                [5 ]Departamento de Química, Universitat Politècnica de València, Camino de Vera s/n, Valencia 46022, Spain
                [6 ]Instituto de Conservación y Mejora de la Agrodiversidad Valenciana, Universitat Politècnica de València, Camino de Vera 14, Valencia 46022, Spain; fraherga@ 123456upvnet.upv.es (F.J.H.); jprohens@ 123456btc.upv.es (J.P.)
                Author notes
                [* ]Correspondence: rmaez@ 123456qim.upv.es ; Tel.: +34-96-3877343
                [†]

                These authors contributed equally to this work.

                Article
                materials-09-00325
                10.3390/ma9050325
                5503040
                © 2016 by the authors;

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license ( http://creativecommons.org/licenses/by/4.0/).

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