Pendahuluan

Nanotechnology has emerged as a promising area of research owing to its wide range of applications in the eld
of biology and medicine. Although various methods are available for the synthesis of metal nanoparticles, most
of the methods such as chemical reduction of metal ions in aqueous solutions (LizMarzan and Lado-Tourino
1996), use of reverse micelles for chemical and photo reduction (Sun et al. 2001), thermal decomposition in
organic solvents (Esumi et al. 1990), and chemical reduction using radiation (Henglein 1998) involve noxious
chemicals. These methods apart from being expensive pose a major threat to environment and also prove to be
dangerous for living beings. Thus, the growing quest to develop eco-friendly nanoparticles led to the
development of biomimetic approach. Biomimetic is an interdisciplinary area involving the synthesis of
nanoparticles using biological principles (Forough and Farhadi 2010).
Synthesis of nanoparticles using various biological approaches including use of enzymes (Willner et al. 2006),
microbes (Klaus et al. 1999; Nair and Pradeep 2002), fungus (Vigneshwaran et al. 2007), plant, or plant extracts
(Chandran et al. 2006; Jae and Beom 2009) have been successfully demonstrated as alternatives to noxious
chemical or physical methods. Notably, the biological synthesis of nanoparticles using plants has received much
more attention in the recent years as it is cost-effective and also overcome the drawback of maintaining pure
microbial cultures as is required in microbial methods. Thus, green synthesis is the recent method implemented
for largescale synthesis of nanoparticles (Iravani 2011).
Free radicals are generated in living systems as part of metabolic activities, and their levels are regulated by
cellular antioxidant system. Uncontrolled levels of reactive oxygen species can cause cell membrane damage
and DNA mutation, which further result in development of many diseases such as cancer, brain disorders, and
cardiovascular diseases (Percival 1998). Cancer remains as a huge threat to public health for developed as well
as developing countries. Despite modern advancements in diagnosis, prevention, and therapy, cancer still affects
millions of people worldwide and is one of the leading causes of death (Ferlay et al. 2004). Many naturally
occurring and synthetic compounds have been studied for their anticancer properties which include antioxidants
(Reed 1999). Many plant-derived phytocompounds
have been shown to possess antioxidant property (Sudjaroen 2009). Metal nanoparticles prepared from plant
extracts have been reported to exhibit considerable antioxidant properties which can be exploited in the
treatment of diseases like cancer (Kanipandian and Thirumurugan 2014; Nakkala et al. 2014).
Gymnema sylvestre has been used in the treatment of diabetes since ages in folk, ayurvedic, and homeopathic
systems of medicine (Grover et al. 2002). It is also used in the treatment of asthma, eye complaints, family
planning, snakebite, urinary complaints, stomach ailments, piles, chronic cough, breathing troubles, colic pain,
cardiopathy, constipation, dyspepsia and hemorrhoids, and hepatosplenomegaly (Patel et al. 2012). Reports on
the antioxidant activity of G. sylvestre are available (Tiwari et al. 2014).
In this study, silver and gold nanoparticles were synthesized using G. sylvestre leaf extract and evaluated for
their in vitro free radical quenching activity and cytotoxic effects in Hep2 cells. Characterization of synthesized
G. sylvestre silver (GYAgNPs) and gold (GYAuNPs) nanoparticles was carried out using different techniques
such as transmission electron microscopyenergy-dispersive X-ray spectroscopy (TEMEDX), Fourier
transform infrared spectroscopy (FT-IR), dynamic light scattering (DLS) particle size analyzer, and
thermogravimetric analysisdifferential scanning calorimetry (TGADSC). The antioxidants activities of
GYAgNPs and GYAuNPs were assessed by different in vitro assays. Further, the cytotoxic effect in Hep2 cells
was evaluated by 3-(4,5dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and induction of
apoptosis-mediated cell death was conrmed by cytomorphology, acridine orange and ethidium bromide
(AO/EB), 2 dichlorouorescin diacetate (DCF-DA), propidium iodide (PI), and 4,6-diamidino-2 phenylindole
dilactate (DAPI) staining techniques