Biofunctionalised Magnesium Oxide nanoparticle using Zerumbone and its Anticancer

activity on (HT29) Colon Cancer Cell Line

Y.J Jeba Jane Ratney*, Dr.S. Begila Davida
*&a
Department of Chemistry, Scott Christian College, Nagercoil.
Dr. R. Subramanianb
b
PSN College of Engineering and Technology, Tirunelveli.
Ferdin.bjf@gmail.com
Abstract
Green synthesis of nanoparticles supports reuse, reduce and recycle concept towards
the environmental friendly synthesis and also produce the good results compare to chemical
methods in some of the cases. Present work concentrated on to union of MgO nanoparticle by
Green technique utilizing zerumbone, were subjected to sub-atomic docking ponders for the
restraint of leukotriene A4 hydrolase (LTA4H) protein is viewed as a significant focus for
disease treatment. In silico expectation utilizing a docking approach uncovered that LTA4H
may be a potential focus of MgO np utilizing Zerumbone. It stifles safe haven autonomous
tumor cell development by restraining LTA4H action in HT 29 colorectal disease cells. The
incorporated Magnesium oxide nanoparticles have been described by structural, optical,
Thermal and indicated powerful cytotoxic impact against HT29 colon growth cell line with in
IC50 estimation of ------ g/ml by the MTT assay. The present study shows the green
incorporated Magnesium oxide nanoparticle is utilized significant higher anticancer
movement. From the outcomes got it is recommended that the green combined MgO
nanoparticle which is a compelling anticancer specialist.
Keywords: Leukotriene A4 Hydrolase; In Silico; Zerumbone; Cytotoxic; MTT Assay
Introduction
Nanomaterials are part of a commercial revolution that has resulted in an explosion of
hundreds of new products due to their diverse physico-chemical properties, enabling their
usage in a wide range of innovative applications [1]. Noble metal nanoparticles have been the
subject of intense research due to their unique optical, electronic, mechanical, magnetic and
chemical properties that are significantly different from those of bulk materials. These
properties are attributed to their small sizes and large surface areas. Preparation of silver
nanoparticles has attracted particularly considerable attention due to their diverse properties
and uses [2] Nanomaterials are important materials due to their novel characteristics, and
unique physical and chemical properties. Certain identified nanoparticles have anticancer
activity through the accumulation of the nanoparticles in the cell from the enhanced
permeability and retention effect.[3]
Nanoparticles can be synthesized using various approaches including chemical,
physical, and biological methods. Although chemical method of synthesis requires short
period of time for synthesis of large quantity of nanoparticles, this method requires capping
agents for size stabilization of the nanoparticles. Chemicals used for nanoparticles synthesis
and stabilization are toxic and lead to non-ecofriendly byproducts. The need for
environmental non-toxic synthetic protocols for nanoparticles synthesis leads to the
developing interest in biological approaches which are free from the use of toxic chemicals as
byproducts. Thus, there is an increasing demand for green nanotechnology[4]
ZnO nanoparticles with a unique optical, electrical, and thermal performance have
been widely used in the field of catalysis, sunscreen cosmetics, paint materials, and food
packaging materials [5,6]. The chemical and physical properties of nanoparticles have a
strong influence on the way they interact with biological components or the environment.[7]
Cancer is an abnormal type of tissue growth in which the cells exhibit and uncontrolled
division leading the increase in the number of dividing cells [8]. Several studies have
reported that silver nanoparticles have significantly induced cell necrosis or apoptosis in

many cell types [9]. There is increasing demands for anticancer therapy [10]. Invitro
cytotoxicity testing procedures reduces the use of laboratory animals [11] and hence use of
cultured tissues and cells have increased [2]. The discovery and identification of new
antitumor drug with low side effects on immune system has become an essential goal in
many studies of immuno-pharmacology [13].
Zingiber officinale Roscoe, belongs to Zingiberaceae family with leafy stem up to
60 cm. Ginger contains a number of different pungent and active ingredients. The rhizome is
rich in the secondary metabolites such as phenolic compounds (gingerol, paradol and
shogaoal), volatile sesquiterpenes (zingiberene and bisabolene) and monoterpenoids
(curcumene and citral) [14] The major pungent compounds found in ginger are the gingerols
[15,16]. 6-gingerol [5-hydroxy-1-(4`-hydroxy- 3`-methoxyphenyl)-3-decanone] has been
shown to have an antipyretic, antitussive, hypotensive [17], cardiotonic [18], antiplatelet
[19], antiangiogenic [20], anti-inflammatory and analgesic [21], cytotoxic, apoptotic [22],
antitumor [23], anticancer [24], antioxidant [25], antihepatotoxic [26], antifungal [27],
vanilloid receptor agonistic [28], cholagogic [29] and antiemetic [30] activities.
The purpose of the present work was the study of molecular docking with
LTA4H protein and invitro anticancer activity of zinc oxide nanoparticle on human colon
cancer cell ine.
Materials & Methods
Green synthesis of Magnesium oxide nanoparticles using zerumbone from Zingiber
Zerumbet
Magnesium acetic acid derivation, NaOH and dish sets was bought from Sigma
Aldrich. All crystal was washed with sterile refined water and dried in a hot air stove before
utilize. The technique for the amalgamation was referenced from writing (Gunalan
Sangeethaa et al, 2012). To the 50 ml of zerumbone from alpinia calcarata, magnesium acetic
acid derivation was included and set under an attractive stirrer. Following 15 minutes NaOH
was included till the pH 12 brought about a pale white fluid arrangement. This was then set in
an attractive stirrer for 2 hours. The pale white ppt was then taken out and washed again and
again with refined water took after by ethanol to get free contaminations. The pale white
powder of MgO nanoparticle was gotten in the wake of drying at 6000 C at 3 hours.
3. Leukotrienes

Fig.1leukotriene protien
The leukotrienes make a class out of fundamentally related paracrine hormones got
from the oxidative digestion system of arachidonic corrosive and are ensnared in human
tumor and endless aggravation [16, 17]. Leukotrienes are found at abnormal states in most
incendiary injuries and are included in the physiologic changes that are normal for the fiery

procedure [18]. LTA4H was appeared to show large amounts of protein expression in specific
sorts of diseases, and its restraint prompts diminished growth occurrence in creature models
[19,20].
4.Anticancer activity of [6]-Gingerol mediated Magnesium Oxide nanoparticle
The anticancer movement of tests on HT 29 cells were dictated by the MTT examine.
Cells (1 105/well) were plated in 0.2 ml of medium/well in 96-well plates. Hatch at 5 %
CO2 hatchery for 72 hours. At that point, include different groupings of the examples in 0.1%
DMSO for 48hrs at 5 % CO2 hatchery. After expulsion of the specimen arrangement and
washing with phosphate-cushioned saline (pH 7.4), 20l/well (5mg/ml) of 3-(4,5-dimethyl-2thiazolyl)- 2,5-diphenyl-tetrazolium bromide (MTT) in phosphate-supported saline
arrangement was included. After 4hrs brooding, 1ml of DMSO was included. Practical cells
were controlled by the absorbance at 540nm. Estimations were performed and the fixation
required for a half hindrance of reasonability (IC50) was resolved graphically. The impact of
the specimens on the multiplication of HT 29 cells were communicated as the % cell
practicality, utilizing the accompanying recipe
HT 29 cells were expressed as the % cell viability, using the following formula:
Calculation
RESULT&DISSCUSSION
% cell viability = A540 of treated cells / A540 of control cells 100

XRD
Figure 2 demonstrates X-beam diffraction examples of MgO nanoparticle blended by
the response of zerumnbone from Zingiber zerumbet rhizome with Magnesium acetic acid
derivation and sodium hydroxide. The pinnacles got from this green strategy is (001),(100),
(101),(102),(110),(111),(103),(201) planes of MgO nanoparticles are in excellent record with
the estimations of standard card (2003 JCPDS NO=89-7746 global community for
Diffraction information). The normal particles size was around 22 nm which was plausible by
Debye Scherrer's condition (D=K/cos )
Counts
Jeba-B
800

600

400

200

0
30

40

50

60

70

Position [2Theta] (Copper (Cu))

Fig-2-----X-Ray diffraction of MgO np by green technique utilizing Zerumbone

Fig 3 (A,B,C,D) demonstrates the surface morphologies of green blended Magnesium oxide
nanoparticle of various amplification by the SEM (JOEL-JSM 6390) machine .Images
demonstrated that the circular molded individual particles and also number of
conglomeration. However the normal size of the chose singular particles are in the request of
44 nm for green strategy.

Fig 3 (A,B,C,D) SEM pictures of MgO np by green technique utilizing zerumbone

Figure 4 demonstrates TEM picture and chose region electron diffraction examples of
Zno np by green technique utilizing Zerumbone from Zingiber zerumbet. Nearness of the
arrangement of rings containing spots in SAED affirmed MgO nps have bigger grain
estimate. It indicates crystalline structure. The crystallinity of MgO np is affirmed by the
XRD and TEM . This spectra has given the data with respect to the way of the gem faces. The
gem planes of MgO np saw by TEM picture.

Fig 4 TEM pictures of MgO np by green technique utilizing Zerumbone

UV-Vis range used to reval the optical property of green orchestrated MgO
nanoparticle utilizing Zerumbone by double beam spectrophotometer (2202). .It can be seen
from the fig that there was concentrated ingestion in the ultra violet band of 200-600nm. The
ingestion wave length at around 298 nm of MgO np recommended the excitonic character at
room temperature.

Fig 5 UV spectra of MgO np by green technique utilizing Zerumbone

Fig 6 demonstrates the FTIR spectra of green synthsised MgO nanoparticle by Bruker
Alpha T (Made in Germany) machine. The crest at 3811 cm -1 can be credited to the OH
extending in the precious stone structure of magnesium hydroxide nanoparticle. The wide
band at 3448cm-1 is ascribed to surface retains OH aggregate. The crest at 2509 cm -1 could

be relegated to CH extending vibration. The top at 1444 cm -1 signifies N-H gather. The crest
at 1384 cm-1 demonstrates the vibration of methylene gathering. The crest at 1008 cm-1
compares to extending vibration of alcohols. The crest at 864 and 686 cm-1 relegated to
sweet-smelling bunches. As indicated by Awwrd et al., there was a crest at 430 connected
MgO nanoparticle.

Fig 6 FT-IR spectra of MgO np by green technique utilizing zerumbone

Anticancer activity
Zerumbone contains a higher measure of bioactive mixes, for example, phenolic
mixes Cytotoxicity of the silver nanoparticles was assessed in vitro against Ht29cell line.
Cytotoxic examination of the example demonstrated an immediate measurements reaction
relationship and cytotoxicity expanded at higher focuses. The specimen exhibited an
extensive cytotoxicity against Ht29cell line. The focus important to create half passing rate
was 25 g/mL. Anticancer exercises of the silver nanoparticles assessed against Ht29cell line
are introduced in Table1 and Fig. 7 . Be that as it may, this zerumbone has been broadly
considered for different anticancer reviews; no work has been completed on the MgO NPs
union and anticancer movement against HT29 cell line. In view of this reason we chose just
zerumbone interceded MgO NPs in the anticancer reviews. MgO NPs demonstrated an
unmistakable cytotoxic impact on HT29 cells and a reasonable concentrationresponse
relationship

Fig 7 Anticancer activity of MgO np by Zerumbone

Tab 1 Anticancer activity of MgO np by Zerumbone

S.No
1
2
3
4
5
6

CONCLUSIONS

Concentration
g/ml
200
100
50
25
12.5
Control

Dilution
Neat
1:1
1:2
1:4
1:8
-

Absorbance
540nm
0.00
0.21
0.34
0.43
0.52
0.52

% cell Viability
0.10
26.02
38.64
62.0
78.0

MgO nanoparticles were combined effectively by Biological technique utilizing zeumbone. The UVVis spectroscopic review affirmed the lessening of metal particle and development of nanoparticle
with crest at - nm. The SEM investigation affirms the arrangement of ZnO nanoparticles with circular
size of distance across running - . The XRD ponder affirms the structure of ZnO nanoparticles and the
development of thin pinnacle proposes the crystalline way of MgO nanoparticles. FTIR estimations
were done to distinguish the conceivable biomolecules in charge of topping and effective adjustment
of the metal nanoparticles integrated by zerumbone. Inside the confinements of this review, the MTT
examine comes about recommend that when the grouping of green integrated MgO nanopowders was
25mg/mL, the feasibility of HT29 cells dropped to around 58%. In this way this review effectively
exhibits the advantageous usage of bio energizes to orchestrate MgO nanopowders and their
conceivable organic applications.

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