Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 137 (2015) 250–258

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Spectrochimica Acta Part A: Molecular and

Biomolecular Spectroscopy
journal homepage: www.elsevier.com/locate/saa

Rambutan peels promoted biomimetic synthesis of bioinspired zinc

oxide nanochains for biomedical applications
R. Yuvakkumar a, J. Suresh b, B. Saravanakumar c, A. Joseph Nathanael d, Sun Ig Hong a,⇑, V. Rajendran c,⇑
a
Department of Nanomaterials Engineering, Chungnam National University, Daejeon 305-764, South Korea
b
Department of Chemistry, Anna University College of Engineering, Kanchipuram 631 552, Tamil Nadu, India
c
Center for Nanoscience and Technology, K.S. Rangasamy College of Technology, Tiruchengode 637 215, Tamil Nadu, India
d
Department of Nano, Medical and Polymer Materials, Yeungnam University, Gyeongsan 712-749, South Korea

h i g h l i g h t s g r a p h i c a l a b s t r a c t

 ZnO synthesis via zinc-ellagate

complex formation.
 Bioinspired ZnO effect on HepG2 liver
cancer cells were explored.
 60% and 40% cell viability was lost
in 7 and 4 days incubated ZnO treated
cells.
 50% and 10% morphological change
was observed in 7 and 4 days ZnO
treated cells.

a r t i c l e i n f o a b s t r a c t

Article history: A naturally occurring rambutan peel waste was employed to synthesis bioinspired zinc oxide nanochains.
Received 9 April 2014 Rambutan peels has the ability of ligating zinc ions as a natural ligation agent resulting in zinc oxide
Received in revised form 12 July 2014 nanochains formation due to its extended polyphenolic system over incubation period. Successful forma-
Accepted 7 August 2014
tion of zinc oxide nanochains was confirmed employing transmission electron microscopy studies. About
Available online 28 August 2014
60% and 40% cell viability was lost and 50% and 10% morphological change was observed in 7 and 4 days
incubated ZnO treated cells compared with control. Moreover, 50% and 55% of cell death was observed at
Keywords:
24 and 48 h incubation with 7 days treated ZnO cells and hence alters and disturbs the growth of cancer
New strategy
Zinc oxide
cells and could be used for liver cancer cell treatment.
Rambutan peel waste Ó 2014 Elsevier B.V. All rights reserved.
Nephelium lappaceum L.
Liver cancer cell treatment

Introduction bility of designing smart bio-nanomaterials with specific biological

Interactions between biological systems and nanostructured
materials are of attracting increasing interest because of the possi-
⇑ Corresponding authors. Tel.: +82 42 8216595; fax: +82 42 8225850 (S.I. Hong).
Tel.: +91 9994130303; fax: +91 4288 274880 (direct), 274860 (V. Rajendran).
E-mail addresses: sihong@cnu.ac.kr (S.I. Hong), veerajendran@gmail.com (V.
Rajendran).
functionalities. There has been an explosive and strategic develop-
ment in novel green syntheses protocols which produces nanoscale
biocompatible nanoparticles and they have been used to probe
biological processes and diagnose biomedical applications [1–3].
In the emerging field of nanobiotechnology, plant extract based
synthetic strategy is one of the most eco-friendly alternative meth-
ods to synthesise nanoparticles [4–6]. In recent times, non-toxic,
biomimetic, environment friendly approaches have been gaining

http://dx.doi.org/10.1016/j.saa.2014.08.022
1386-1425/Ó 2014 Elsevier B.V. All rights reserved.
 R. Yuvakkumar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 137 (2015) 250–258 251

Fig. 1. XRD, Raman, PL, XPS pattern: (a, d, g, j, m, p) 1, (b, e, h, k, n, q) 4, (c, f, i, l, o, r) 7 days of reaction product from the mixture of 0.1 M Zn(NO3)2
6H2O and 10 mL extract.

more importance. Their ability to form a wide range of biocompat-
ible nanomaterials paves the way to best suited for biomedical
applications [7–13]. Moreover, green synthesis of nanoparticles
played a vital role and gaining much attention due to its simplicity,
inexpensive and eco-friendly nature. Biofunctional ZnO nanomate-
rials have been obtained by using several green synthesis proce-
dures. For example, Nagajyothi and co-workers designed a green
route to biosynthesis of ZnO nanoparticles using Ptelea trifoliata
fruit extract and investigated the catalytic activity [7]. Rajiv and
co-workers employed bio-fabrication of zinc oxide nanoparticles
using Parthenium hysterophorus L. leaf extract and studied its size-
dependent antifungal activity against plant fungal pathogens [8].
Selvarajan and co-workers portrayed a novel method for the bio-
synthesis of ZnO nanoparticles using a probiotic bacteria Lactobacil-
lus plantarum VITES07 [9]. Jayaseelan and co-workers described a
low-cost, simple procedure for novel microbial route to synthesize
ZnO nanoparticles using Aeromonas hydrophila as eco-friendly
reducing and capping agent and studied their activity against path-
ogenic bacteria and fungi [10]. In a separate study, Sangeetha and
co-workers employed green synthesis of zinc oxide nanoparticles
by Aloe barbadensis miller leaf extract [11]. Moreover, biocompati-
ble and bioinspired nanomaterials is of particular importance for
various biomedical applications [14–17].
However, transition metal oxide nanostructures synthesis
through inorganic complex intermediate using green ligation
agents obtained from natural resources is one of the emerging
thrust areas of interest in green nanomaterials research today.
Recently, this concept in green nanomaterials research has gained
252 R. Yuvakkumar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 137 (2015) 250–258
Fig. 2. Mechanism for ZnO nanoparticles formation.
substantial interest in the research community [18,19]. In general,
the synthesis of zinc oxide nanoparticles has garnered substantial
interest due to their biomedical applications [20–22]. Additionally,
ZnO nanoparticles are known to have good antibacterial properties
to suit biomedical applications [23–26]. In the present study, a
novel biomimetic synthesis of bioinspired zinc oxide nanoparticles
in the presence of rambutan extract polyphenol have been studied
at room temperature incubated at 1, 4 and 7 days. This is particu-
larly advantageous because this method utilizes rambutan waste
as a resource material as well as a natural ligation agent for inno-
vative biomedical applications [27–32]. Moreover, the discarded
rambutan fruit peels contain powerful phenolic antioxidants [33].
Rambutan peels principal components such as ellagic acid, corila-
gin, geranin and ellagitannins are responsible for the potential uti-
lization in both food and medical industry. Rambutan peel extract
also possesses antibacterial activity against pathogenic bacteria
[34,35]. Therefore, we explored whether the formed bioinspired
ZnO had enhanced biomedical applications. We found that the bio-
inspired ZnO had substantially enhanced HepG2 liver cancer cell
treatments when compared to the control.
Experimental details
Preparation of ZnO
Rambutan fruit exported from Thailand was collected from the
Daejeon super market, South Korea. Zinc nitrate [Zn(NO3)2
6H2O]
and ethanol were purchased from Merck chemicals. Rambutan
peels were washed with running water and subsequently incised
into small pieces and placed in circulating oven at 50 °C until
complete dryness. 3 g peels were boiled with ethanol and double
distilled water (1:2 ratio) for 10 min and filtered through What-
man No. 1 filter paper. 0.1 M Zn(NO3)2
6H2O was prepared in
50 mL and 10 mL extract was slowly added drop wise under mag-
netic stirring at 80 °C for 2 h and then incubated at room temper-
ature for 1, 4 and 7 days to form zinc-ellagate complex formation.
Zinc-ellagate complex formed after adequate time of incubation
was directly decomposed in muffle furnace at 450 °C in static air
atmosphere.
Characterization of ZnO
Phase and crystalline nature of prepared samples were identi-
fied by X-ray powder diffraction patterns (Bruker AXS, Germany)
using Cu Ka as the radiation (1.5406 Å) source. Average crystallite
size of zinc oxide samples was calculated using Scherrer’s formula.
High-resolution (Dispersive Raman microscope, Horiba Jobin Yvon;
LabRAM HR UV/Vis/NIR) Raman spectroscopy (excitation, 514 nm)
was used to confirm ZnO. X-ray photoelectron spectra (XPS) were
obtained using a MultiLab 2000 spectrometer (Thermo Electron
Corporation, UK) to identify the elements present in the samples.
The HPLC spectrum was recorded employing LC-10AD pump,
SPD-10A UV/Vis detector and SIL-10A auto sampler (Shimadzu).
Infrared (IR) spectra were recorded using a Fourier transform infra-
red spectrophotometer using a Nicolet 520P FT-IR spectrometer.
Thermogravimetric analysis (TGA) was performed on a Perkin
Elmer 7 Series thermal analysis system under a nitrogen flow
(50 mL/min). The surface morphology of the product was recorded
using scanning electron microscopy (S-3000H; Hitachi, Japan). An
elemental analysis, energy-dispersive X-ray spectroscopy, was
 R. Yuvakkumar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 137 (2015) 250–258 253

Fig. 3. HPLC: (a) Pure (b) 1, (c) 4, (d) 7 days of reaction product from the mixture of 0.1 M Zn(NO3)2
6H2O and 10 mL extract.

Fig. 4. FTIR, TGA pattern: (a and d) 1, (b and e) 4, (c and f) 7 days of reaction product from the mixture of 0.1 M Zn(NO3)2
6H2O and 10 mL extract.

performed to evaluate the composition of the sample along with Liver cancer cell application
the scanning electron microscopy observation. Transmission elec-
tron microscopic images were obtained using a transmission elec- In order to gain insight into the morphology of liver cancer cells,
tron microscope (CM 200; Philips, USA). 8–10 mm in diameter of 4 and 7 days treated ZnO were incubated
254 R. Yuvakkumar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 137 (2015) 250–258
with HepG2 liver cancer cells. The cells were harvested by trypsin-
ization and centrifugation after the incubation period. The cell pel-
let was suspended in phosphate buffered saline (PBS) with 0.1%
tryphan blue. The morphology of the stained cells was observed
using an inverted microscope. For cell counting, 2  103 cells were
loaded on 4 and 7 days treated ZnO and then Dulbecco’s Modified
Eagle Medium (DMEM) medium with 10% Fetal Bovine Serum
(FBS) was added; the cells were then incubated at 37 °C with 5%
CO2. At intervals of 12, 24 and 48 h, the cells were harvested by
trypsinization and centrifugation after the incubation period. The
obtained cell pellets were suspended in PBS with 0.1% tryphan blue
and then stained cells were counted using a light microscope. For
cell proliferation, 2  104 cells were seeded in 24-well plates and
incubated with 4 and 7 days treated ZnO at different time intervals.
Cell proliferation was assayed using the MTT assay (3-(4,5-Dimeth-
ylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) method by tak-
ing the optical density of viable cells in 570 nm in an Enzyme
Linked Immuno Sorbent Assay (ELISA) reader (Bio-Rad, USA).
Results and discussion
Characterization of materials
The bioinspired ZnO nanostructure using rambutan peel extract
was demonstrated and confirmed by the characteristic peaks
observed in the XRD image. The XRD pattern of the obtained prod-
uct was shown in Fig. 1a–c. The narrow and strong diffraction
peaks revealed that the obtained product were pure and well crys-
talline in nature. The XRD peaks were identified as 100, 002, 101,
102, 110, 103, 112 and 201 reflections respectively in the whole
spectrum of 2 theta values of 31, 34, 36, 47, 56, 63, 66, 68, 69, 72
and 76. All of the diffraction peaks can be indexed to the spherical
and hexagonal wurtzite structure phase of zinc oxide by compari-
son with the data from JCPDS card No. 89-7102 and no other char-
acteristic peaks were observed than ZnO. A similar result was also
observed in Calotropis procera [12]. The average crystallite size was
obtained from the XRD peaks using Scherrer’s formula applied to
the major intense peaks [36]. The calculated average particle diam-
eter was found to be in the range of about 50 nm. Line broadening
of the diffraction peaks is an indication that the synthesized mate-
rials are in the nanometer range.
Fig. 1d–f shows the Raman spectra of ZnO. The ZnO optical
modes at 99, 380, 410, 438 and 583 was respectively corresponded
high
to Elow
2 , A1 (Transverse Optical), E1 (Transverse Optical), E2 and E1
(Longitudinal Optical) first order optical phonons [37]. The addi-
tional modes observed in ZnO at 205, 331, 536 and 659 cm 1 (M1
to M4) were assigned respectively to overtones and combinations
high
corresponding to 2Elow 2 , E2 Elow
2 , 2LA (Longitudinal Acoustic)
and TA + LO (Transverse Acoustic + Longitudinal Optical) phonons
[38]. As seen in Fig. 1g–i, an increase in absorbance and a shift from
380 nm to 750 nm in the peak were observed in PL spectra as time
progressed. The absorbance spectrum of the ZnO is consistent with
the highly categorized surface plasmon resonance for nanosized
ZnO present at 380 nm. The peak at 536 nm also indicates the for-
mation of largely hexagonal ZnO consistent with the TEM images.
1, 4 and 7 days incubated ZnO samples showed a typical near-
band-gap UV emission at 380 nm and a broad visible emission
band centered at around 530 nm which is usually attributed to
deep-level emission caused by defects of oxygen vacancies. Gener-
ally, nanocrystalline ZnO exhibits 2 peaks in PL spectrum around
380 and 500 nm. Red shift emission suggests increase in incuba-
tion time increases particle sizes. The XPS survey spectrum and
high resolution XPS spectra of the prepared ZnO are shown in
Fig. 1j–r. The survey XPS spectrum shown in Fig. 1j–l clearly
revealed the Zn and O peaks and concluded that the synthesized
product was ZnO. Binding energies around 1023.13 and
1046.23 eV shown in Fig. 1m–o were attributed to Zn 2p3/2 and
2p1/2 electrons respectively. The lone and sharp peak at
1023.13 eV for the Zn 2p3/2 core level is associated with the Zn spe-
cies in the completely oxidized state [39]. The intense peak at
531.7 eV shown in O1s spectrum (Fig. 1p–r) of the prepared ZnO
could be attributed to ZnO oxygen.
Incubation of ZnO
ZnO nanostructures formation scheme in presence of rambutan
peel extract is shown in Fig. 2. Ellagic acid, gallic acid and tannic
acid, polyphenols, phenolic derivatives like dihydroxy benzene,
presence of hydroxyl groups, ellagic acid of plant polyphenolic com-
pounds have been utilized to form stable complexes with metal ions
such as copper, zinc and several other metal ions [40–44]. The uti-
lized rambutan extract may have an affinity towards zinc ions due
to its extensive polyphenolic ring system and may consequently
form zinc-ellagate complexes and undergo direct decomposition
at 450 °C leads to zinc oxide nanostructures. HPLC was explored
to study rambutan peel extract phenolic compounds role in ZnO
synthesis (Fig. 3a–d). Pure, 1, 4 and 7 days of reaction product from
the mixture of 0.1 M Zn(NO3)2
6H2O and 10 mL extract showed a
well defined peak (retention time) at 6.4 min corresponds to the
presence of ellagic acid [45]. Pure rambutan extract without the zinc
source showed a highest peak (Fig. 3a). Moreover, 1, 4 and 7 days of
reaction product also showed a definite peak (Fig. 3b–d). This clearly
revealed and confirmed the attachment of the extract on the
obtained product. The attachment of the extract on the surface of
the obtained nanoparticles was also confirmed later employing
TEM studies. As shown in Fig. 3a–d, the additional peaks observed
at 8, 13 and 14 min (Geraniin Peak; 10–15 min) in the HPLC spec-
trum were most likely due to the major compound in ethanolic
Nephelium lappaceum peel extract [46]. The major compounds
detected in rambutan peel extract analysis were ellagic acid and
geraniin. Moreover, it was observed that the attachment of the
extract was dependent upon the incubation time to form the bioin-
spired zinc oxide. When the incubation time was increased, the
attachment of the rambutan peel extract was increased as evi-
denced from the obtained variation in HPLC peaks. The obtained
HPLC results clearly revealed that the rambutan peel extract binds
to the surface of the ZnO incubated at 1, 4 and 7 days of the reaction
product.
Fig. 4a–c shows the FTIR spectra of rambutan fruit extract pro-
moted ZnO. A strong peak observed in the hydroxyl region of
3300–3500 cm 1 was due to the phenolic AOH groups. The pres-
ence of obtained phenolic AOH groups in IR spectra supported
the possible mechanism of zinc oxide formation using rambutan
peel extract as shown in Fig. 2. The ester oxygen atom and the phe-
nolic hydroxyl groups of polyphenols form p-track conjugation
effect when the hydroxyl groups form binding with the metal as
metal–phenolate complex (Zinc-ellagate complex) by the chelating
effect. The role of the chelating effect will make metal ion more
stable which may be consistent with zinc-ellagate complex forma-
tion. The obtained IR spectra clearly revealed that the polyphenols
and hydroxyl groups were involved in ZnO formation. In addition,
the peaks observed at 1633 cm 1 could be assigned to the C@C
stretch due to the aromatic ring system. The weaker bands
observed at 2753 and 831 cm 1 could be assigned to CAH stretch
(alkanes) and CAH stretch (aromatics) functional groups. The
absorption peaks at 1386 cm 1 can be attributed the CAC stretch-
ing of aromatic rings in the synthesis of ZnO. In our synthesis, the
strong peak of aromatic hydroxyl groups (1633, 1386 and
831 cm 1) present in rambutan peel extracts ligate with zinc ions
to form the zinc-ellagate complex at pH 5. The pH value varies
depending upon the metal ions [47]. In addition, the peak observed
 R. Yuvakkumar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 137 (2015) 250–258
Fig. 5. TEM image: (a–c) 1, (d–f) 4, (g–i) 7 days of reaction product from the mixture of 0.1 M Zn(NO3)2
6H2O and 10 mL extract.
at 2416 cm 1 was assigned to quinoid containing keto–enol sys-
tem. The formation of product most likely occurs due to the two-
electron oxidation of the hydroxyl groups of the phenol ring sys-
tem of the extract as reported elsewhere [48]. The spectrum
showed a single band at 445 cm 1 attributed to metal–oxygen
(MAO) vibrational band. The peak in the region between 400 and
600 cm 1 were allotted to ZnAO [49]. The results obtained from
the IR spectra are having the combined characteristics of zinc oxide
as well as secondary compounds of rambutan extract (Fig. 4a and
b) and hence showing importance of functional groups in ZnO syn-
thesis. HPLC and FTIR analysis confirmed the presence of natural
phenolic antioxidant compounds and hence offer favorable effects
for the synthesis of bioinspired zinc oxide.
Thermal analysis curve (Fig. 4d–f) showed a uniform weight
loss from 200 °C up to about 1000 °C of the bioinspired ZnO using
fruit extracts. A small uniform weight loss occurring from 200 °C
can be ascribed to the elimination of water, ethanol and partial
dehydroxylation of ZnO. Fig. 4d showed a steady minor weight loss
in 7 days incubated sample when compared to 1 and 4 days in the
temperature range of 200–1000 °C due to the elimination of the
adsorbed water and to the dehydroxylation process. The same
has been evidenced from HPLC results (Fig. 3d). The TGA curve of
the 1 and 4 days incubated ZnO also showed steady weight losses
between 200 and 1000 °C attributed to the loss of water from ZnO;
255
as well as differentiated behavior marked by a uniform weight loss
due to minor attachment of the extract at low incubation time. It
was found that the prepared ZnO maintained good thermal stabil-
ity and 7 days incubated samples was little more stable when com-
pared to 1 and 4 days incubated ZnO samples. When incubation
time increases, the attachment of the extract on the bioinspired
sample was increased. According to the results obtained from
HPLC, FTIR and TGA, it was clear that the attachment of the rambu-
tan extract plays a key role in the formation of bioinspired ZnO. It
may be due to the rambutan extracts which possess ellagic acid
which readily attracts the cations and triggers ZnO synthesis.
Moreover, all these factors cumulatively support the synthesis of
bioinspired ZnO in the presence of rambutan peel extract.
In addition, TEM was employed to characterize the surface
attachment of the extract and to confirm the size, shape and mor-
phology of the bioinspired ZnO as shown in Fig. 5a–i. From the TEM
images (Fig. 5a–i), it is clear that the morphology of the bioinspired
ZnO was to be spherical and hexagonal and the size ranges
between 50 and 100 nm and also some of the particles were
agglomerates. As shown in Fig. 5a–i, when incubation time
increased, the sizes of the product increased and different shapes
of nanoparticles such as spherical and hexagonal were observed
(Fig. 5a–i). After 4 days, we observed the formation of multidimen-
sional chain-like structures in which spherical nanoparticles were
256 R. Yuvakkumar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 137 (2015) 250–258
Fig. 6. SEM–EDX pattern: (a and b) 1, (c and d) 4, (e and f) 7 days of reaction product from the mixture of 0.1 M Zn(NO3)2
6H2O and 10 mL extract.
intertwined to each other (Fig. 5d and e, g and h). These figures
undoubtedly indicate the obtained particles to be spherical and
hexagonal structure. The SAED images of the prepared ZnO nano-
particles at 10(1/nm) scales are shown in Fig. 5(c, f, and i) respec-
tively. The electronic diffraction pattern (Fig. 5c, f, and i) confirmed
the formation of the highly crystalline zinc oxide nanostructure
with (1 0 0), (1 0 1) and (1 1 0) phases. It was observed that the ram-
butan extract was mainly appearing to attach on the surface of bio-
inspired ZnO as shown in Fig. 5e–h. This is very similar to the
synthesis of bioinspired zinc oxide nanoparticles as reported else-
where [10]. The FESEM images also exhibit distinguished spherical
and hexagonal morphology as shown in Fig. 6a, c, and e. The syn-
thesized ZnO nanoparticles were generally in random and not uni-
form, which is very similar to those described in the previous
literature [8]. The EDX (energy-dispersive X-ray) shows the chem-
ical composition for the synthesized ZnO. Fig. 6(b, d, and f) shows
the EDX spectrum recorded in the spot-profile mode from one of
the densely populated ZnO nanoparticles area. Fig. 6b, d, and f rep-
resents the strong signals from the zinc and oxygen atoms and
other signals from C atoms were also observed at low angle which
is very similar to Jayaseelan et al. [10]. The signals were likely due
to X-ray emission from the rambutan extract attached in the
surface of the bioinspired ZnO and this may be due to the role of
ellagic acid adopted to attach with the nanoparticles.
Owing to all the results that observed, the presence of rambutan
extract ellagic acid appears to attach the surface of the bioinspired
ZnO nanoparticles. In the past, such behavior has been observed in
the presence of ellagic acid [41]. Thus, in the presence of rambutan
extract, initially different sizes and shapes of ZnO nanoparticles are
formed and the interactions with rambutan extract as an attach-
ment allow biomedical favorable application due to the presence
of the polyphenolic systems and could applicable to liver cancer
cell treatments. It appears that rambutan extract not only ligating
zinc ions and also attach in the surface of bioinspired zinc oxide
nanoparticles. All the obtained results clearly revealed that
rambutan extract binds to the ZnO leading to promote possible
biomedical applications. Moreover, the researchers have been
investigating the possibility of using the finely tuned ZnO for the
development of antiproliferative nanohybrid compound with con-
trolled release property using ellagic acid as the active agent [50].
Moreover, in the present study, obtained ZnO at room temperature
in the presence of rambutan ellagic acid could be more suitable for
biomedical applications. Therefore, the obtained nanoparticles
were examined for HepG2 liver cancer cell treatments.
 R. Yuvakkumar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 137 (2015) 250–258
Fig. 7. Cell morphology (a) control, (b) 4 days incubated ZnO treated cells and (c) 7 days incubated ZnO treated cells, bar: 10 lm.
Fig. 8. Cell counting: control, 4 and 7 days incubated ZnO treated cells.
G2 liver cancer cells test
Zinc oxide nanoparticles are potential candidate for biomedical
cancer applications and therapeutic treatment [50] and gaining
interest due to their high stability [51], inherent photolumines-
cence [52], photocatalytic systems [53] and promotion of reactive
oxygen species generation [54]. The shape and morphology such
as spherical, one-dimensional, tetrapod-like ZnO nanomaterials is
another important consideration for biomedical applications. It is
possible to enhance permeation and retention effect due to the
enhanced physico chemical characteristics such as increased sur-
face to volume ratio and reactivity by tailoring morphology and
size [1]. When nanoparticles interact with cells, the reactive oxy-
gen species (ROS) generation induced and hence lead to cell death
257
when antioxidative capacity of the cell is exceeded [54]. Enhanced
in vivo anti-tumor efficacy has been developed employing nanom-
aterials as pharmaceutical carriers for delivery and diagnostics
agents. ZnO is also a promising therapy to arrest in vivo tumor
growth and to kill cancer cells employing UV, magnetic field or
shortwave radiofrequency energy irradiated. When sufficient
supernormal temperatures are reached, the tumor cells are killed
without harming surrounding healthy tissue. Recent improve-
ments have been described by conjugating cell specific ligands to
the surface of nanoparticles resulting in greater control of drug tar-
geting at the tissue and cellular levels and by encapsulating drugs
within nanoparticles [1]. Therefore, in the present work, the bioin-
spired 4 and 7 days treated ZnO at low incubation temperature
were used for liver cancer cell treatment and the status of the cells
is explored for cell morphology and cell proliferation studies along
with control as shown in Fig. 7a–c. Initially, 2  103 cells were
loaded and the total cell count in a neubauer chamber was
observed. The number of viable cell counts was reduced in 4 and
7 days treated ZnO as shown in Fig. 8 when compared to control.
More than 60% and 40% of cell viability was lost in 7 and 4 days
ZnO treated cells. 7 days ZnO treated cells were 50% modified from
normal cells and were irregular and morphologically affected
(Fig. 7c) and 4 days treated ZnO cells showed 10% morphological
change (Fig. 7b). Therefore, in the present study, 7 days treated
ZnO alters and disturbs the growth of cancer cells. The maximum
inhibition of cell proliferation was observed at 24 and 48 h of incu-
bation. Nearly 50% and 55% cell death was observed at 24 and 48 h
incubation with 7 days treated ZnO. Moreover, 4 days treated ZnO
showed not very significant cell death compared with 7 days
treated ZnO. Thus, the studies on ZnO treated at 4 and 7 days cells
suggest that 7 days treated ZnO can be used as a more suitable bio-
material in liver cancer cell treatment. In conclusion, bioinspired
ZnO by a new and green cost-effective method has been explored
258 R. Yuvakkumar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 137 (2015) 250–258
in the presence of the rambutan peel extract. The formation of bio-
inspired ZnO in the presence of rambutan extract has an added
advantage of being attached to ZnO could have potential biomed-
ical applications.
Conclusions
In this study, rambutan extract assisted biomimetic syntheses
of zinc oxide nanochains with different incubation times were
explored. The topographical analysis confirmed the attachment of
biomaterial on the surface of the obtained product. The effects of
nano ZnO on G2 liver cancer cells were studied. 60% and 40% cell
viability was lost respectively in 7 and 4 days ZnO treated cells and
50% and 10% cell morphological change was observed in 7 and
4 days ZnO treated cells. About 50% and 55% cell death was
observed at 24 and 48 h incubation in 7 days treated ZnO. The pre-
pared 7 days treated bioinspired ZnO with a well-defined nano-
structure at lower period of incubation could be used in liver
cancer cell treatments.
Acknowledgment
This work was supported by Chungnam National University
Research Fund (2014).
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