International Journal of Biological Macromolecules 138 (2019) 1064–1071

Contents lists available at ScienceDirect

International Journal of Biological Macromolecules

journal homepage: http://www.elsevier.com/locate/ijbiomac

Surface-modified nanocrystalline cellulose from oil palm empty fruit

bunch for effective binding of curcumin
Mei Ling Foo a, Ca Rol Tan a, Pei Dee Lim a, Chien Wei Ooi a, Khang Wei Tan b,c,⁎, Irene Mei Leng Chew a,⁎
a
School of Engineering, Monash University Malaysia, 47500 Bandar Sunway, Selangor, Malaysia
b
School of Energy and Chemical Engineering, Xiamen University Malaysia, Selangor Darul Ehsan 43900, Malaysia
c
College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China

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

Article history: Rod-shape particles have been a good drug carrier due to the long circulatory time, tumor accumulation and high
Received 15 December 2018 cellular uptake in body. Acid-hydrolysed nanocrystalline cellulose (NCC) from empty fruit bunch exhibited a
Received in revised form 10 June 2019 width of 13–30 nm and a length of 150–360 nm in rod-shape structure. NCC holds good potential as a bio-
Accepted 4 July 2019
based drug carrier owing to its biodegradability and biocompatibility. Fourier-transform infrared spectroscopy
Available online 10 July 2019
results confirmed the binding of curcumin onto the NCC modified with tannic acid (TA) and decylamine (DA).
Keywords:
TA-DA modification rendered NCC with a higher level of hydrophobicity, as evidenced by a substantial increase
Nanocrystalline cellulose in contact angle from 45° to 73°. The modified NCC had the curcumin-binding efficiency in the range of 95–99%,
Empty fruit bunch which is at least twofold higher than the unmodified NCC at any curcumin concentration tested. This remarkable
Hydrophobic curcumin-binding effciency was comparable to that of commercialized NCC from wood-based origin. This work
Surface modification suggests NCC as a superior and sustainable drug carrier, while TA-DA modification is a promising approach to
Curcumin alter the surface property of NCC for an efficient binding of curcumin.
Drug carrier © 2019 Elsevier B.V. All rights reserved.

1. Introduction functionalization of its surface via different surface chemistry [18–21],

Short circulatory half-life and poor bioavailability of water-insoluble
drug (e.g. anticancer drugs) have always been the barriers to an effec-
tive delivery of drug in human body [1,2]. To improve the therapeutic
efficacy of drug, drug carrier including liposomes [3], polymeric nano-
particle [4,5], nanogel [6,7], micelles [8] and emulsion [9,10] have
been widely explored. These carriers improved the drug solubility and
prolonged drug's circulation time in human body at a lower adminis-
tered dose [11,12]. Typically, most of these carriers were designed in
the form of spheres due to the ease of manufacturing process [1]. Stud-
ies suggested that rod-shape particles, such as nanorods of gold [13,14],
polystyrene [15] and silica [16], would be able to improve the blood
clearance kinetics, cellular uptake of drug, and tumor accumulation
[1,17]. Nevertheless, the potentially high cost of materials (e.g. gold
nanorod) and the controversial safe administration of synthetic inor-
ganic matters demonstrated the need for other inexpensive and biolog-
ically benign options.
Nanocrystalline cellulose (NCC), a nanomaterial with rod-shaped
structure, shows great potential as a favorable drug carrier. Its high sur-
face area and abundance of surface hydroxyl groups allow
⁎ Corresponding authors.
E-mail addresses: khangwei.tan@xmu.edu.my (K.W. Tan), irene.chew@monash.edu
(I.M.L. Chew).
facilitating a high level of drug loading [22,23]. Moreover, the successful
extraction of NCC from a broad range of lignocellulosic biomass, such as
wood [24], bagasse [25,26], kenaf [27,28], roselle fiber [29], seaweed
[30], and oil palm biomass [31–33] has greatly demonstrated its prom-
ising renewability and sustainability. Empty fruit bunch (EFB), accounts
for 22% (dry basis) per ton of fresh fruit bunches processed in palm oil
industry. Currently, EFB is mainly used in the low-value applications
such as organic mulch in plantation and supplementary fertilizer. De-
spite its potential in biofuel production, the energy-intensive feedstock
preparation, high capital and operating costs remained challenging to
raise its competitiveness against fossil fuel [34]. The high volume of un-
utilized EFB has therefore become a promising source for NCC produc-
tion. Numerous efforts on diverse approaches of NCC extraction
[33,35,36] have been attempted. The potential utilization of EFB-
derived NCC as drug carrier, however, was rarely explored.
In the present work, an approach of surface modification based on
tannic acid (TA) and decylamine (DA) was adapted from Hu et al. [37]
to alter the surface properties of EFB-derived NCC. NCC with the modi-
fied hydrophobic surface was then tested for its efficiency in binding
curcumin, a type of water-insoluble drug. Curcumin is a naturally occur-
ring polyphenol renowned for its outstanding pharmacological features
[38] including anti-inflammatory [39–41], antioxidant [3], antimicrobial
[42], anticancer [43,44], and antimutagenic [10,11] properties. Similar
to other types of water-insoluble drugs, the selection of carriers is

https://doi.org/10.1016/j.ijbiomac.2019.07.035
0141-8130/© 2019 Elsevier B.V. All rights reserved.
 M.L. Foo et al. / International Journal of Biological Macromolecules 138 (2019) 1064–1071
crucial to the efficiency of curcumin delivery [11,45]. Hence, the binding
capacity of curcumin onto NCC with and without surface modification
was evaluated to confirm the viability of TA and DA as a substitution
to the commonly adopted surfactant, cetyl trimethylammonium bro-
mide (CTAB), which might interact with the phospholipid bilayers of
cells and lead to cell death [22].
To our knowledge, this is the first work presenting EFB-derived NCC
tailored with hydrophobic surfaces for curcumin binding. The
curcumin-binding efficiency of NCC was quantified spectrophotometri-
cally using UV–vis spectrometer. The Fourier-transform infrared spec-
troscopy and field-emission scanning electron microscopy were used
to examine the binding of curcumin onto NCC surface. The present
work exploited the morphological, structural and biological (i.e. bio-
compatibility, biodegradability) advantages of NCC to enhance its po-
tential uses in nano-carrier aspect.
2. Materials and methods
2.1. Materials
EFB fibers were the complimentary samples given by Eureka Syn-
ergy (Malaysia). Sodium hydroxide and sulfuric acid (H2SO4, 95–97%)
were purchased from Friendemann Schmidt Chemical. Sodium chlorite
(80%), tannic acid (95%), n-Decylamine (99%) were supplied by Acros
Organics. Glacial acetic acid (AR grade) and curcumin (95% from Tur-
meric rhizone) were obtained from Fisher Scientific and Alfa Aesar, re-
spectively. Ethanol (absolute grade) was purchased from Merck.
Wood-origin NCC (NCCwood) with 5–20 nm width and 150–200 nm
length was produced by Process Development Center, University of
Maine. All reagents were of analytical grade and used without further
purification. All aqueous solutions were prepared with deionized (DI)
water (N18.2 MΩ×cm resistivity).
2.2. Preparation of NCC from EFB
EFB fibers were first ground using a rotor mill (Pulverisette 14,
Fritsch) and filtered through a 250-μm siever. To remove the water-
soluble substances, the fibers were washed with water at 50 °C. The fi-
bers were then dried in an oven at 60 °C to a constant weight. An acidic‐
sodium chlorite solution made of equal volume of 2 w/v% of NaClO2 and
acetate buffer (mixture of NaOH and CH3COOH at a defined pH) was
used in the delignification of fibers. The fibers were added into the
pre-heated acidic sodium chlorite solution at 80 °C and stirred for 2 h.
The delignified fibers were then treated with 4 w/v% NaOH to degrade
the hemicellulose at 80 °C for 2 h under constant stirring. Removal of
chemical residual on the fibers surface was done through water wash-
ing at the end of each delignification and alkali treatment. The washed
pulp was filtered with Whatman filter paper (pore size = 11 μm) and
was then subjected to freeze drying. The freeze-dried fibers were sub-
jected to acid hydrolysis using 58 wt% of H2SO4 for 45 min at 45 °C.
Upon completion of acid hydrolysis, cold water was added into the
acid solution to stop the process. The mixture was then centrifuged at
10,000 rpm for 15 min for the collection of hydrolysed sample. This
step was repeated twice for complete removal of acid residue. The sam-
ple was dialyzed using a dialysis membrane (molecular weight cut off
= 12,000–14,000 Da) against deionized water until a constant pH was
achieved. The collected solid suspension was then treated with sonica-
tion (Q700, QSONICA) at 60% amplitude for 10 min to obtain the dis-
persed NCC.
2.3. Modification of nanocrystalline cellulose
The method of modifying NCC using TA and DA was adapted from an
earlier work [37]. NCC suspension (c.a. 1 w/v%) was adjusted to pH 8
using NaOH solution. TA was first added to the NCC suspension at the
concentration of 1 mg/mL. The mixture was stirred for 6 h at room
1065
temperature to form NCCTA suspension. Then, DA was added to the
NCCTA suspension at the concentration of 40 mg/mL. The resulted
NCCTA-DA suspension was stirred for 3 h, before being centrifuged for
10 min at 1000 rpm. The supernatant was discarded and the pellet
was re-suspended in DI water followed by another round of centrifuga-
tion. This step was repeated twice. Finally, the modified EFB-derived
NCC suspension, (NCCTA-DA) was collected. The wood-origin NCC
(NCCwood-TA-DA) prepared using the abovementioned modification pro-
cedures was used as the control.
2.4. Curcumin loading
A stock solution of curcumin was first prepared in ethanol at a con-
centration of 100 μg curcumin/mL ethanol. The stock solution was then
diluted to a desired concentration, and the NCC sample (c.a. 1 mg/mL)
was added to the curcumin solution. The mixture was tumbled at
9 rpm for 60 min using a rotisserie tube rotator at room temperature.
The resulted suspension was subjected to centrifugation to pelletize
the curcumin-bound NCC. The concentration of unbound curcumin in
the supernatant was assayed to determine the binding efficiency.
2.5. Characterizations
2.5.1. X-ray diffraction (XRD) analysis
The crystallinity of samples was analyzed using an XRD diffractome-
ter (AXS-D8, Bruker). The diffracted intensity of the Cu Kα radiation was
measured in the range of 2θ = 5–50° at a scan rate of 0.03° s−1. The em-
pirical method [46] was exploited to estimate the crystallinity index, CrI
(%), on the basis of reflected intensity data by using Eq. (1):
I002 −Iam
CrI ð%Þ ¼  100 ð1Þ
I 002
where I002 and Iam are the intensities of peak in the crystalline and
amorphous regions, respectively.
2.5.2. Thermogravimetric analysis (TGA)
TGA was performed using a thermogravimetric analyser (Q50, TA In-
struments) to determine the degradation characteristics of the cellulose
samples. The samples were heated from room temperature to 600 °C at
a heating rate of 10 °C/min under nitrogen gas flow rate of 60 mL/min.
2.5.3. Fourier-transform infrared spectroscopy (FTIR)
The infrared spectra of samples were obtained using a FTIR spectro-
photometer (Thermo-Nicolet iS10, Thermo Fisher Scientific) equipped
with ATR diamond probe accessory. The spectra were collected in the
transmittance mode from an accumulation of 64 scans over the range
of 400–4000 cm−1.
2.5.4. Contact angle measurement
The contact angles of NCC samples were measured using a goniom-
eter (Ramehart) equipped with a computer-automated program. The
suspension was deposited on a microscope glass plate followed by
oven-drying. With the aid of image analysis tool, the contact angle of
water on the coated glass plate was determined based on sessile drop
technique.
2.5.5. Measurement of particle size and ζ-potential
The hydrodynamic diameters and ζ-potential of samples were mea-
sured by a Zetasizer (Nano-ZS, Malvern) based on dynamic light scatter-
ing (DLS) method.
2.5.6. Microscopy analysis
Prior to microscopy analysis, the NCC suspension was diluted and
dispersed in DI water, whereas curcumin-bound NCCTA-DA was dis-
persed in ethanol. The surface morphology of NCC and curcumin-
1066 M.L. Foo et al. / International Journal of Biological Macromolecules 138 (2019) 1064–1071
bound NCCTA-DA were observed using a field-emission scanning electron
microscope (FE-SEM; SU8010, Hitachi) operated at scanning transmis-
sion electron microscopy (STEM) mode at 15 kV. The diluted suspension
(~ 0.001 w/v%) was dropped onto a carbon-coated copper grid and was
then air-dried at room temperature. To enhance the contrast, the NCC
was stained with a drop of 2 w/v% uranyl acetate followed by air-
drying. The particles size in the microscopic image was estimated
using ImageJ software.
2.5.7. Curcumin-binding analysis
The unbound curcumin in the supernatant of curcumin-bound NCC
pellet was quantified spectrophotometrically with a UV–vis spectrome-
ter (Cary 100, Agilent) at 428 nm at ambient temperature. The concen-
tration of curcumin was determined from a calibration plot (R2 = 0.99)
prepared using different concentrations of curcumin (1–60 μg/mL). The
curcumin-binding efficiency was calculated as follows:
Binding efficiency ð%Þ
Curcumin added−unbound curcumin
¼  100
Curcumin added
2.5.8. Statistical analysis
Analysis of variance (ANOVA) was performed using GraphPad Prism
5 software at the significance level of p b 0.05.
3. Results & discussion
3.1. Characterization of NCC
3.1.1. XRD analysis
As shown in Fig. 1, the XRD patterns of raw EFB, purified fiber and
EFB-derived NCC indicated the predominance of Cellulose I structure
in their compositions. The sharp crystalline peaks observed at around
2θ = 22.6° and 34.6°, as well as the broad amorphous humps at around
14.7° and 16.2° revealed the typical diffractograms of semi-crystalline
material [46–48]. After pre-treatment of EFB, the CrI of the purified fi-
bers increased significantly from 38.8% to 66.1%, which was primarily
ascribed to the effective removal of amorphous lignin and hemicellu-
lose. The dissolution of amorphous domains and hydrolytic cleavage
of glycosidic bonds during acid hydrolysis released the individual crys-
tallites [49,50] and led to an increase in CrI. Thus, a higher value of CrI
Fig. 1. X-ray diffraction patterns of NCC, purified fiber, and EFB.
(77.6%) was obtained from the acid-hydrolysed NCC. The intensified
crystalline peaks detected at 22.6° indicated a higher degree of crystal-
line perfection in the (0 0 2) plane, as a result of the progressive extrac-
tion of cellulose. A higher degree of crystallinity also led to a clearer
distinction between (1 0 1) and (1 0 1) planes, corresponding to the
characteristic peaks in amorphous domains of cellulose [51].
3.1.2. Thermogravimetric analysis
The thermal behaviors of NCC samples were examined using TGA,
and the results are presented in Fig. 2. The initial weight loss observed
below 100 °C was due to the water evaporation. As observed in Fig. 2,
the decomposition of EFB and purified fiber occurred at 280 °C and 293
°C, respectively. The relatively lower decomposition temperature of EFB
fiber was ascribed to the early degradation of pectin, lignin and hemicel-
lulose in the EFB as compared to that of cellulose [52,53]. Upon heating to
400 °C, the oxidation/decomposition of solid residue took place and re-
leased the gaseous products with a lower molecular weight [33,55].
The non-cellulosic components of EFB fiber contributed to a higher quan-
ð2Þ tity of char residue at 600 °C as compared to the purified fiber [55]. On
the other hand, the gradual weight loss of NCC at and above 200 °C
was attributed to the degradation of sulfated cellulose. The sulfate
group, which was introduced during the acid hydrolysis, could decrease
the activation energies of degradation, catalyzing the decomposition of
NCC at lower temperature prior to the acid hydrolysis treatment
[56,57]. Furthermore, a higher content of char residue in NCC is usually
anticipated upon completion of heating as compared to the raw and pu-
rified fibers; this was attributed to the flame retardancy property of NCC
[55,57]. The thermal characteristics of these cellulose samples were pri-
marily governed by the structural changes caused by the extraction con-
dition, leading to the different trend of thermal decomposition.
3.2. Characterization of modified NCC
3.2.1. Fourier-transform infrared spectroscopy
FTIR was used to confirm the structural changes in NCC at different
stages of extraction, surface modification and curcumin loading. The
FTIR spectra are shown in Fig. 3. The ester bond between hydroxyl of lig-
nin and carboxyl of uronic acid in hemicellulose was disrupted during
the alkali treatment; the acid-chlorite delignification potently cleaved
the ether bond between lignin and cellulose [58,59]. Referring to
Fig. 3e, the peaks at 1505 cm−1 and 1592 cm−1 were attributed to the
aromatic skeletal vibration in lignin [60–62]. In addition, the peak lo-
cated at 1729 cm−1 corresponded to C\\O stretching vibration of the
EFB NCC Purified Fiber
100
90
80
70
Weight loss (%)
60
50
40
30
20
10
0
0 100 200 300 400 500 600
Temperature (°C)
Fig. 2. Mass loss curves from thermogravimetric analysis of the EFB, purified fiber and NCC.
 M.L. Foo et al. / International Journal of Biological Macromolecules 138 (2019) 1064–1071
Fig. 3. Fourier-transform infrared spectroscopy spectra of (a) curcumin-bound NCCTA-DA, (b) NCCTA-DA, (c) NCC, (d) purified EFB fiber, and (e) raw EFB.
acetyl and uronic ester groups presented in the ester linkage of carbox-
ylic group of lignin or/and hemicellulose [26,63]. The disappearance of
these characteristic peaks of lignin and hemicellulose indicated the suc-
cess removal of lignin and hemicellulose from the fibers after the steps
of delignification and alkali treatment. The peak observed at around
897 cm−1 was ascribed to C\\H rocking vibration of carbohydrate asso-
ciated with β-glycosidic linkage of cellulose [25,64,65]. Similarly, the
bands in the region of 1420–1430 cm−1, which are usually observed
in the spectra of polysaccharides, were attributed to the symmetric
bending of CH2, as well as the bending of C\\H and C\\O groups in aro-
matic rings [27]. A peak at 1204 cm−1 corresponding to the sulfate
groups found in NCC samples suggested that the celluloses had under-
gone the esterification reaction during acid hydrolysis treatment [56].
It is noteworthy that the peaks located at 897 cm−1 and 1427 cm−1
are the characteristic infrared bands of Type I cellulose [65] and are in
good agreement with the results obtained from XRD analysis. In general,
the FTIR results obtained at different stages of extraction corresponded
well with their respective XRD patterns. In particularly, the increase in
crystallinity of NCC indicated the effective removal of amorphous poly-
mers (i.e. hemicellulose and lignin) and the success formation of NCC.
As shown in Fig. 3a and b, the broad bands in the 2800–2900 cm−1 re-
gion are commonly observed in the surface-modified cellulosic mate-
rial, in which the asymmetrical and symmetrical stretching of CH2 in
C10 alkyl chain were observed at both 2850 cm−1 and 2919 cm−1 in
the spectrum [37,66]. The notable changes across the broad band in
3200–3500 cm−1 region were also observed. This region primarily
corresponded to the O\\H stretching vibration in cellulose molecules
[25], while the prominent peaks around 3300 cm−1 and 3331 cm−1
were ascribed to the N\\H stretching [67]. Furthermore, a new peak at
1457 cm−1 corresponding to C\\C aromatic stretching vibration and
an intense peak at 1644 cm−1 assigned to C_N stretching [68] were de-
tected in Fig. 3b, evidently suggesting the attachment of DA on NCC. The
Fig. 4. Contact angle of (a) NCC, (b) NCCTA, (c) NCCTA-DA.
1067
prominent peak at 1277 cm−1 in FTIR spectrum of NCCTA-DA, which cor-
responds to the C\\H stretching vibration, indicated the successful
binding of curcumin onto NCCTA-DA [19].
3.2.2. Contact angle analysis
To evaluate the wetting behaviors of modified and unmodified NCC
samples, the contact angle analysis was conducted and the results are
shown in Fig. 4. As seen in Fig. 4a, the contact angle of unmodified NCC
was 45°, indicating the hydrophilic surface properties of native NCC. In
contrast, the NCC modified with TA (NCCTA) had a smaller surface con-
tact angle (34°, see Fig. 4b), while the NCCTA coupled with DA (NCCTA-
DA) had a larger contact angle (73°, see Fig. 4c). Generally, the hydropho-
bic surface typically leads to a contact angle N90°, however, for ease of
understanding, the resulted NCCTA-DA was defined as hydrophobic due
to its water-insoluble behavior. The TA was found to have depressing ef-
fect to the resulting contact angle of NCCTA [37,69], which indicated the
increased hydrophobicity of NCCTA-DA was chiefly contributed by DA.
3.2.3. DLS analysis
The equivalent hydrodynamic diameter and surface charge of NCC
were determined to understand the effect of surface modification on
the properties of NCC. Fig. 5 shows the z-average size and ζ-potential
of NCC. The z-average sizes of unmodified NCC and NCCTA-DA were
75 nm and 882 nm, respectively. Such a remarkable increment (p b
0.05) in size of NCCTA-DA was attributed to the formation of water-
insoluble large aggregates triggered by the hydrophobic nature of DA
[37]. NCC is known to be amphiphilic but its hydrophilic character is
more dominant due to the protruding hydroxyl groups [70,71]. It is pos-
tulated that the hydroxyl groups on NCC's surface are the potential sites
for the interaction between TA and NCC, forming the precursor for sub-
sequent attachment of DA. Despite the mechanism of TA deposition is
yet to be fully unraveled, the five-polyphenol-arm structure of TA is
1068 M.L. Foo et al. / International Journal of Biological Macromolecules 138 (2019) 1064–1071

Fig. 5. Z-average size and ζ-potential of NCC, NCCTA and NCCTA-DA measured using dynamic light scattering analysis. Different letters (a to d) indicate statistically significant differences (p b
0.05) between groups (means ± standard deviation) of triplicate measurements.

known to possess diverse bonding functionalities. It was suggested that
the TA anchored onto NCC surface primarily via its catechol/pyrogallol-
type phenol through oxidation under alkaline condition [72,73]. TA
tends to dissociate at a higher pH and becomes negatively charged
[74]. Oxidation of TA leads to the formation of unstable intermediate
known as quinones, which is highly reactive as the electrophilic entities
for attachment of different compounds [72,75,76]. Essentially, the TA
deposition was a bio-inspired concept emerged from the adhesive prop-
erties of protein found in mussel [75,77]. The nucleophilic amines attack
the reactive quinones and engage in secondary reaction, forming the
high-molecular-weight hydrophobic adducts [37,78,79]. The increased
ζ-potential (p b 0.05) of NCCTA-DA implied the neutralization of nega-
tively charges of dissociated TA [74], which could be attributed to the
stable covalent coupling [37,42,80] between the amino moieties of DA
and quinone. Furthermore, the occurrence of intermolecular interac-
tions such as hydrogen-bonding and π-stacking between quinone and
NCC (or quinone molecules), was also speculated [72]. Thus, these prin-
cipal reactions and intermolecular interaction occurred during TA-DA
modification are believed to be responsible for the hydrophobic surface
properties in NCCTA-DA.
the unmodified NCC and NCCwood-TA-DA. The results are shown in
Fig. 8. The curcumin-binding efficiency of unmodified NCC was found
to be a function of curcumin concentration, with the curcumin-
binding efficiency increased from 8% to 54% when the amount of loaded
curcumin increased from 20 μg/mL to 100 μg/mL (p b 0.05 for 40 to 100
μg/mL). The hydrophobic characteristic of cellulose was contributed by
its hydrophobic C\\H bonds located at the axial position of the glucopy-
ranose ring [70,71]. Thus, the hydrophobic interaction between
curcumin and C\\H moieties of cellulose was induced, allowing the
curcumin to be bound to the unmodified NCC in a curcumin-
concentration-dependent manner. The binding capacity of NCCTA-DA
was significantly increased (p b 0.05) and was approximately two-fold
higher than NCC, regardless of the range of curcumin concentration
tested. We infer that the increased level of hydrophobicity in NCCTA-DA
favored the hydrophobic interaction between the hydrophobic phenolic
moieties of curcumin and the long alkyl chain of DA, resulting in the

3.3. Characterization of curcumin-bound surface modified-NCC

3.3.1. Microscopy analysis

The STEM images of EFB-derived NCC and curcumin-loaded NCCTA-DA
are presented in Figs. 6 and 7, respectively. The width and length of the
rod-shaped NCC were estimated to be 13–30 nm and 150–360 nm
(Fig. 6), respectively. The aspect ratio of NCC dimension was as high as
27, indicating a high surface area of NCC available for drug loading
[22]. This high-aspect-ratio feature allowed a large quantity of curcumin
particles to be dispersed onto NCC surface, leading to an effective binding
of curcumin onto NCC wherein the shape and structure of modified NCC
remained unchanged (Fig. 7).

3.3.2. Curcumin-binding analysis

The curcumin-binding efficiency of NCCTA-DA with curcumin loaded
at 20, 40, 60, 80 and 100 μg/mL was examined and compared against Fig. 6. Scanning transmission electron microscopy image of NCC.
 M.L. Foo et al. / International Journal of Biological Macromolecules 138 (2019) 1064–1071 1069

Fig. 7. Scanning transmission electron microscopy image of curcumin-bound NCCTA-DA with 100 μg/mL of curcumin dispersed in ethanol.

remarkable curcumin-binding efficiency ranging from 95% to 99%. Fur-
thermore, we also surmise that the excellent binding capacity of
NCCTA-DA could be ascribed to the emergence of entanglement of
NCCTA-DA and the formation of crosslinked-like network, facilitating
the binding of curcumin particles onto the surface of NCCTA-DA. Eventu-
ally, the curcumin-bound NCCTA-DA attracted more curcumin molecules
towards the network as the tumbling process proceeded. The similar
binding behavior was also observed by Zainuddin et al. [66] who used
CTAB-modified NCC for curcumin binding; a higher binding efficiency
was achieved when a higher amount of curcumin was loaded to the
CTAB-modified NCC suspension. We speculate that this phenomenon
could possibly be attributed by the increased number of active sites on
curcumin molecular structure, in which both hydrogen bonding and hy-
drophobic interaction are the predominant contributing factors. Fig. 9
shows the possible mechanism for the binding of curcumin onto the
surface of NCC modified using TA and DA through Schiff base and Mi-
chael addition [37,72,73,78]. Generally, the chemical structure of
curcumin contains several potential sites of attachment for different
carriers [38,81]. Hydrogen bonding is an important force involved in
the binding of curcumin onto NCC. The phenolic hydroxyl and methoxy
groups on curcumin's molecular structure and the center enol groups
are the potential hydrogen donors, while the ketone group located in
the middle acts as hydrogen acceptor [38]. NCCwood-TA-DA was used as
a control to benchmark the curcumin-binding efficiency; the efficiency
of curcumin binding onto NCCwood-TA-DA was in the range of 95–98%.
As shown in Fig. 8, the performance of both NCCTA-DA and NCCwood-TA-
DA in curcumin-binding tests was comparable (p b 0.05 for 40 to 100
μg/mL), indicating that EFB-derived NCC is a promising candidate for
making the concept of waste-to-wealth into a reality. Moreover, surface
modification using TA and DA was proven to be a feasible approach for
tailoring NCC as an effective drug carrier.
4. Conclusion
NCC with 13–30 nm width and 150–360 nm length was successfully
extracted from EFB. The rod-shaped NCC exhibits Cellulose I structure
with CrI of 77.6%. After the surface modification, the formation of stable
covalent coupling between TA and DA in NCC resulted in a greater

Fig. 8. Curcumin-binding efficiency of NCC, NCCTA-DA and NCCwood-TA-DA at different curcumin concentration. Average value of triplicate measurements was presented as means ± standard
deviation. Different letters (a to i) indicate statistically significant differences (p b 0.05) between means.
1070 M.L. Foo et al. / International Journal of Biological Macromolecules 138 (2019) 1064–1071
Fig. 9. Schematic illustration of the possible mechanism for binding of curcumin onto surface of NCC modified using TA and DA.
hydrophobicity with a contact angle of 73°. The binding capacity of hy-
drophobic NCC and its potential as a carrier for curcumin were evalu-
ated. The STEM image confirmed the binding of curcumin onto NCC.
The NCCTA-DA achieved a remarkable efficiency in curcumin binding
(99%) as compared to the maximum binding of curcumin (54%) onto
the unmodified NCC. Moreover, the curcumin-binding efficiency of
NCCTA-DA presented in this work was comparable to that of the commer-
cialized NCCwood-TA-DA. This revealed that the NCCTA-DA is an effective
and promising carrier for hydrophobic particles. Furthermore, this find-
ing also signified the potential of tapping EFB as the abundant source of
NCC for application as the drug carrier. Nonetheless, future studies shall
be conducted in the aspects of in vivo cytotoxicity, drug stability, tumor-
targeting properties and eventually proof of concept to assess the safety
and efficacy of NCCTA-DA in cancer treatment.
Acknowledgement
The authors would like to acknowledge the financial support from
Monash University Malaysia (Higher Degree by Research Scholarships)
and the Ministry of Higher Education, Malaysia (FRGS/2/2014/TK05/
MUSM/03/1).
References
[1] N.P. Truong, M.R. Whittaker, C.W. Mak, T.P. Davis, The importance of nanoparticle
shape in cancer drug delivery, Expert Opin. Drug Deliv. 12 (1) (2015) 129–142.
[2] Y.A. Luqmani, Mechanisms of drug resistance in cancer chemotherapy, Med. Princ.
Pract. 14 (Suppl. 1) (2005) 35–48.
[3] M. Takahashi, S. Uechi, K. Takara, Y. Asikin, K. Wada, Evaluation of an oral carrier sys-
tem in rats: bioavailability and antioxidant properties of liposome-encapsulated
curcumin, J. Agric. Food Chem. 57 (19) (2009) 9141–9146.
[4] N. Sanoj Rejinold, M. Muthunarayanan, V.V. Divyarani, P.R. Sreerekha, K.P.
Chennazhi, S.V. Nair, H. Tamura, R. Jayakumar, Curcumin-loaded biocompatible
thermoresponsive polymeric nanoparticles for cancer drug delivery, J. Colloid Inter-
face Sci. 360 (1) (2011) 39–51.
[5] K.W. Tan, S.Y. Tang, T. Renjan, N. Vasanthakumari, M. Sivakumar, Curcumin-loaded
sterically stabilized nanodispersion based on non-ionic colloidal system induced by ul-
trasound and solvent diffusion-evaporation, Pure Appl. Chem. 88 (1–2) (2016) 43–60.
[6] S.S. Bansal, M. Goel, F. Aqil, M.V. Vadhanam, R.C. Gupta, Advanced drug-delivery sys-
tems of curcumin for cancer chemoprevention, Cancer Prev. Res. 4 (8) (2011)
1158–1171.
[7] J.K. Oh, D.I. Lee, J.M. Park, Biopolymer-based microgels/nanogels for drug delivery
applications, Prog. Polym. Sci. 34 (12) (2009) 1261–1282.
[8] G. Gaucher, M.-H. Dufresne, V.P. Sant, N. Kang, D. Maysinger, J.-C. Leroux, Block co-
polymer micelles: preparation, characterization and application in drug delivery, J.
Control. Release 109 (1) (2005) 169–188.
[9] S. Ganta, H. Devalapally, M. Amiji, Curcumin enhances oral bioavailability and anti-
tumor therapeutic efficacy of paclitaxel upon administration in nanoemulsion for-
mulation, J. Pharm. Sci. 99 (11) (2010) 4630–4641.
[10] S. Ganta, M. Amiji, Coadministration of paclitaxel and curcumin in nanoemulsion
formulations to overcome multidrug resistance in tumor cells, Mol. Pharm. 6 (3)
(2009) 928–939.
[11] M.M. Yallapu, M. Jaggi, S.C. Chauhan, Curcumin nanoformulations: a future
nanomedicine for cancer, Drug Discov. Today 17 (1) (2012) 71–80.
[12] W.-H. Lee, C.-Y. Loo, P.M. Young, D. Traini, R.S. Mason, R. Rohanizadeh, Recent ad-
vances in curcumin nanoformulation for cancer therapy, Expert Opin. Drug Deliv.
11 (8) (2014) 1183–1201.
[13] Arnida, M.M. Janát-Amsbury, A. Ray, C.M. Peterson, H. Ghandehari, Geometry and
surface characteristics of gold nanoparticles influence their biodistribution and up-
take by macrophages, Eur. J. Pharm. Biopharm. 77 (3) (2011) 417–423.
[14] K. Niikura, T. Matsunaga, T. Suzuki, S. Kobayashi, H. Yamaguchi, Y. Orba, A.
Kawaguchi, H. Hasegawa, K. Kajino, T. Ninomiya, K. Ijiro, H. Sawa, Gold nanoparti-
cles as a vaccine platform: influence of size and shape on immunological responses
in vitro and in vivo, ACS Nano 7 (5) (2013) 3926–3938.
[15] S. Barua, J.-W. Yoo, P. Kolhar, A. Wakankar, Y.R. Gokarn, S. Mitragotri, Particle shape
enhances specificity of antibody-displaying nanoparticles, Proc. Natl. Acad. Sci. 110
(9) (2013) 3270.
[16] V.P. Chauhan, Z. Popović, O. Chen, J. Cui, D. Fukumura, M.G. Bawendi, R.K. Jain, Fluo-
rescent nanorods and nanospheres for real-time in vivo probing of nanoparticle
shape-dependent tumor penetration, Angew. Chem. Int. Ed. 50 (48) (2011)
11417–11420.
[17] A. Banerjee, J. Qi, R. Gogoi, J. Wong, S. Mitragotri, Role of nanoparticle size, shape and
surface chemistry in oral drug delivery, J. Control. Release 238 (2016) 176–185.
[18] S. Eyley, W. Thielemans, Surface modification of cellulose nanocrystals, Nanoscale 6
(14) (2014) 7764–7779.
[19] M.M. Yallapu, M.R. Dobberpuhl, D.M. Maher, M. Jaggi, S.C. Chauhan, Design of
curcumin loaded cellulose nanoparticles for prostate cancer, Curr. Drug Metab. 13
(1) (2012) 120–128.
 M.L. Foo et al. / International Journal of Biological Macromolecules 138 (2019) 1064–1071
[20] K. Tan, S. Heo, M. Foo, I.M. Chew, C. Yoo, An insight into nanocellulose as soft con-
densed matter: challenge and future prospective toward environmental sustainabil-
ity, Sci. Total Environ. 650 (1) (2019) 1309–1326.
[21] S. Dong, H.J. Cho, Y.W. Lee, M. Roman, Synthesis and cellular uptake of folic acid-
conjugated cellulose nanocrystals for cancer targeting, Biomacromolecules 15 (5)
(2014) 1560–1567.
[22] A.B. Seabra, J.S. Bernardes, W.J. Fávaro, A.J. Paula, N. Durán, Cellulose nanocrystals as
carriers in medicine and their toxicities: a review, Carbohydr. Polym. 181 (2018)
514–527.
[23] J.K. Jackson, K. Letchford, B.Z. Wasserman, L. Ye, W.Y. Hamad, H.M. Burt, The use of
nanocrystalline cellulose for the binding and controlled release of drugs, Int. J.
Nanomedicine 6 (2011) 321–330.
[24] S. Beck-Candanedo, M. Roman, D.G. Gray, Effect of reaction conditions on the prop-
erties and behavior of wood cellulose nanocrystal suspensions, Biomacromolecules
6 (2) (2005) 1048–1054.
[25] M.R.K. Sofla, R.J. Brown, T. Tsuzuki, T.J. Rainey, A comparison of cellulose
nanocrystals and cellulose nanofibres extracted from bagasse using acid and ball
milling methods, Adv. Nat. Sci. Nanosci. Nanotechnol. 7 (3) (2016) 035004.
[26] A. Mandal, D. Chakrabarty, Isolation of nanocellulose from waste sugarcane bagasse
(SCB) and its characterization, Carbohydr. Polym. 86 (3) (2011) 1291–1299.
[27] M. Jonoobi, J. Harun, M. Mishra, K. Oksman, Chemical composition, crystallinity and
thermal degradation of bleached and unbleached kenaf bast (Hibiscus cannabinus)
pulp and nanofiber, BioResources 4 (2) (2009) 626–639.
[28] H. Kargarzadeh, I. Ahmad, I. Abdullah, A. Dufresne, S.Y. Zainudin, R.M. Sheltami, Ef-
fects of hydrolysis conditions on the morphology, crystallinity, and thermal stability
of cellulose nanocrystals extracted from kenaf bast fibers, Cellulose 19 (3) (2012)
855–866.
[29] L.K. Kian, M. Jawaid, H. Ariffin, Z. Karim, Isolation and characterization of nanocrys-
talline cellulose from roselle-derived microcrystalline cellulose, Int. J. Biol.
Macromol. 114 (2018) 54–63.
[30] S. Singh, K.K. Gaikwad, S.-I. Park, Y.S. Lee, Microwave-assisted step reduced extrac-
tion of seaweed (Gelidiella aceroso) cellulose nanocrystals, Int. J. Biol. Macromol.
99 (2017) 506–510.
[31] M.S. Mohaiyiddin, O.H. Lin, W.T. Owi, C.H. Chan, C.H. Chia, S. Zakaria, A.R. Villagracia,
H.M. Akil, Characterization of nanocellulose recovery from Elaeis guineensis frond
for sustainable development, Clean Techn. Environ. Policy 18 (8) (2016) 2503–2512.
[32] J. Lamaming, R. Hashim, O. Sulaiman, C.P. Leh, T. Sugimoto, N.A. Nordin, Cellulose
nanocrystals isolated from oil palm trunk, Carbohydr. Polym. 127 (2015) 202–208.
[33] A.A. Al-Dulaimi, W.D. Wanrosli, Isolation and characterization of nanocrystalline cel-
lulose from totally chlorine free oil palm empty fruit bunch pulp, J. Polym. Environ.
25 (2) (2017) 192–202.
[34] S.H. Chang, An overview of empty fruit bunch from oil palm as feedstock for bio-oil
production, Biomass Bioenergy 62 (2014) 174–181.
[35] F. Fahma, S. Iwamoto, N. Hori, T. Iwata, A. Takemura, Isolation, preparation, and
characterization of nanofibers from oil palm empty-fruit-bunch (OPEFB), Cellulose
17 (5) (2010) 977–985.
[36] N. Razali, M.S. Hossain, O.A. Taiwo, M. Ibrahim, N.W. Mohd Nadzri, N. Razak, N.F.
Mohammad Rawi, M. Mohd Mahadar, M.H. Mohamad Kassim, Influence of acid hy-
drolysis reaction time on the isolation of cellulose nanowhiskers from oil palm
empty fruit bunch microcrystalline cellulose, BioResources 12 (3) (2017) 6773–6788.
[37] Z. Hu, R.M. Berry, R. Pelton, E.D. Cranston, One-pot water-based hydrophobic surface
modification of cellulose nanocrystals using plant polyphenols, ACS Sustain. Chem.
Eng. 5 (6) (2017) 5018–5026.
[38] M. Salem, S. Rohani, E.R. Gillies, Curcumin, a promising anti-cancer therapeutic: a re-
view of its chemical properties, bioactivity and approaches to cancer cell delivery,
RSC Adv. 4 (21) (2014) 10815–10829.
[39] L.M.G. Zambrano, D.A. Brandao, F.R.G. Rocha, R.P. Marsiglio, I.B. Longo, F.L. Primo,
A.C. Tedesco, M.R. Guimaraes-Stabili, C. Rossa Junior, Local administration of
curcumin-loaded nanoparticles effectively inhibits inflammation and bone resorp-
tion associated with experimental periodontal disease, Sci. Rep. 8 (2018) 6652.
[40] V.R. Yadav, S. Suresh, K. Devi, S. Yadav, Effect of cyclodextrin complexation of
curcumin on its solubility and antiangiogenic and anti-inflammatory activity in rat
colitis model, AAPS PharmSciTech 10 (3) (2009) 752–762.
[41] Q. Chen, S. Gou, Y. Huang, X. Zhou, Q. Li, M.K. Han, Y. Kang, B. Xiao, Facile fabrication
of bowl-shaped microparticles for oral curcumin delivery to ulcerative colitis tissue,
Colloids Surf. B: Biointerfaces 169 (2018) 92–98.
[42] G. Xu, D. Pranantyo, B. Zhang, L. Xu, K.-G. Neoh, E.-T. Kang, Tannic acid anchored
layer-by-layer covalent deposition of parasin I peptide for antifouling and antimi-
crobial coatings, RSC Adv. 6 (18) (2016) 14809–14818.
[43] R.K. Gangwar, G.B. Tomar, V.A. Dhumale, S. Zinjarde, R.B. Sharma, S. Datar, Curcumin
conjugated silica nanoparticles for improving bioavailability and its anticancer ap-
plications, J. Agric. Food Chem. 61 (40) (2013) 9632–9637.
[44] M.M. Yallapu, S. Khan, D.M. Maher, M.C. Ebeling, V. Sundram, N. Chauhan, A. Ganju,
S. Balakrishna, B.K. Gupta, N. Zafar, M. Jaggi, S.C. Chauhan, Anti-cancer activity of
curcumin loaded nanoparticles in prostate cancer, Biomaterials 35 (30) (2014)
8635–8648.
[45] A. Shehzad, F. Wahid, S. Lee Young, Curcumin in cancer chemoprevention: molecu-
lar targets, pharmacokinetics, bioavailability, and clinical trials, Arch. Pharm. 343 (9)
(2010) 489–499.
[46] L. Segal, J.J. Creely, A.E. Martin, C.M. Conrad, An empirical method for estimating the
degree of crystallinity of native cellulose using the x-ray diffractometer, Text. Res. J.
29 (10) (1959) 786–794.
[47] M. Wada, L. Heux, J. Sugiyama, Polymorphism of cellulose I family:reinvestigation of
cellulose IVI, Biomacromolecules 5 (4) (2004) 1385–1391.
[48] Q. Li, S. Renneckar, Supramolecular structure characterization of molecularly thin
cellulose I nanoparticles, Biomacromolecules 12 (3) (2011) 650–659.
1071
[49] M.M. de Souza Lima, R. Borsali, Rodlike cellulose microcrystals: structure, properties,
and applications, Macromol. Rapid Commun. 25 (7) (2004) 771–787.
[50] M.S. Islam, N. Kao, S.N. Bhattacharya, R. Gupta, H.J. Choi, Potential aspect of rice husk
biomass in Australia for nanocrystalline cellulose production, Chin. J. Chem. Eng. 26
(3) (2018) 465–476.
[51] P. Lu, Y.-L. Hsieh, Preparation and characterization of cellulose nanocrystals from
rice straw, Carbohydr. Polym. 87 (1) (2012) 564–573.
[52] J.I. Morán, V.A. Alvarez, V.P. Cyras, A. Vázquez, Extraction of cellulose and prepara-
tion of nanocellulose from sisal fibers, Cellulose 15 (1) (2008) 149–159.
[53] N. Johar, I. Ahmad, A. Dufresne, Extraction, preparation and characterization of cel-
lulose fibres and nanocrystals from rice husk, Ind. Crop. Prod. 37 (1) (2012) 93–99.
[55] W.P. Flauzino Neto, H.A. Silvério, N.O. Dantas, D. Pasquini, Extraction and character-
ization of cellulose nanocrystals from agro-industrial residue – soy hulls, Ind. Crop.
Prod. 42 (2013) 480–488.
[56] P. Lu, Y.-L. Hsieh, Preparation and properties of cellulose nanocrystals: rods, spheres,
and network, Carbohydr. Polym. 82 (2) (2010) 329–336.
[57] M. Roman, W.T. Winter, Effect of sulfate groups from sulfuric acid hydrolysis on the
thermal degradation behavior of bacterial cellulose, Biomacromolecules 5 (5)
(2004) 1671–1677.
[58] Y. Zhou, H. Stuart-Williams, G.D. Farquhar, C.H. Hocart, The use of natural abun-
dance stable isotopic ratios to indicate the presence of oxygen-containing chemical
linkages between cellulose and lignin in plant cell walls, Phytochemistry 71 (8)
(2010) 982–993.
[59] J. Zhang, Y.S. Choi, C.G. Yoo, T.H. Kim, R.C. Brown, B.H. Shanks, Cellulose–
hemicellulose and cellulose–lignin interactions during fast pyrolysis, ACS Sustain.
Chem. Eng. 3 (2) (2015) 293–301.
[60] B. Xiao, X.F. Sun, R. Sun, Chemical, structural, and thermal characterizations of alkali-
soluble lignins and hemicelluloses, and cellulose from maize stems, rye straw, and
rice straw, Polym. Degrad. Stab. 74 (2) (2001) 307–319.
[61] A. Kumar, Y.S. Negi, N.K. Bhardwaj, V. Choudhary, Synthesis and characterization of
methylcellulose/PVA based porous composite, Carbohydr. Polym. 88 (4) (2012)
1364–1372.
[62] P. Garside, P. Wyeth, Identification of cellulosic fibres by FTIR spectroscopy - thread
and single fibre analysis by attenuated total reflectance, Stud. Conserv. 48 (4)
(2003) 269–275.
[63] X.F. Sun, F. Xu, R.C. Sun, P. Fowler, M.S. Baird, Characteristics of degraded cellulose
obtained from steam-exploded wheat straw, Carbohydr. Res. 340 (1) (2005) 97–106.
[64] A. Alemdar, M. Sain, Isolation and characterization of nanofibers from agricultural
residues – wheat straw and soy hulls, Bioresour. Technol. 99 (6) (2008) 1664–1671.
[65] Y.S.N. Anuj Kumar, Veena Choudhary, Nishi Kant Bhardwaj, Characterization of cel-
lulose nanocrystals produced by acid-hydrolysis from sugarcane bagasse as agro-
waste, J. Mater. Phys. Chem. 2 (1) (2014) 1–8.
[66] N. Zainuddin, I. Ahmad, H. Kargarzadeh, S. Ramli, Hydrophobic kenaf nanocrystal-
line cellulose for the binding of curcumin, Carbohydr. Polym. 163 (2017) 261–269.
[67] A. Khan, R.A. Khan, S. Salmieri, C. Le Tien, B. Riedl, J. Bouchard, G. Chauve, V. Tan, M.R.
Kamal, M. Lacroix, Mechanical and barrier properties of nanocrystalline cellulose re-
inforced chitosan based nanocomposite films, Carbohydr. Polym. 90 (4) (2012)
1601–1608.
[68] T. Sreethawong, K.W. Shah, S.-Y. Zhang, E. Ye, S.H. Lim, U. Maheswaran, W.Y. Mao,
M.-Y. Han, Optimized production of copper nanostructures with high yields for effi-
cient use as thermal conductivity-enhancing PCM dopant, J. Mater. Chem. A 2 (10)
(2014) 3417–3423.
[69] T. Güler, E. Polat, Tannic acid as a hydrophobicity modifier for galena in the presence
of metal ions, Physicochem. Probl. Miner. Process. 53 (1) (2016) 5–16.
[70] C. Yamane, T. Aoyagi, M. Ago, K. Sato, K. Okajima, T. Takahashi, Two different surface
properties of regenerated cellulose due to structural anisotropy, Polym. J. 38 (8)
(2006) 819–826.
[71] D.J. Gardner, G.S. Oporto, R. Mills, M.A.S.A. Samir, Adhesion and surface issues in cel-
lulose and nanocellulose, J. Adhes. Sci. Technol. 22 (5–6) (2008) 545–567.
[72] Q. Stéphane, D. Denis, D.-C. Céline, P. Laurent, Plant polyphenols: chemical properties,
biological activities, and synthesis, Angew. Chem. Int. Ed. 50 (3) (2011) 586–621.
[73] J. Guo, W. Sun, J.P. Kim, X. Lu, Q. Li, M. Lin, O. Mrowczynski, E.B. Rizk, J. Cheng, G.
Qian, J. Yang, Development of tannin-inspired antimicrobial bioadhesives, Acta
Biomater. 72 (2018) 35–44.
[74] C. Sun, B. Xiong, Y. Pan, H. Cui, Adsorption removal of tannic acid from aqueous so-
lution by polyaniline: analysis of operating parameters and mechanism, J. Colloid In-
terface Sci. 487 (2017) 175–181.
[75] H. Lee, S.M. Dellatore, W.M. Miller, P.B. Messersmith, Mussel-inspired surface chem-
istry for multifunctional coatings, Science 318 (5849) (2007) 426–430.
[76] C.M.G.C. Renard, A.A. Watrelot, C. Le Bourvellec, Interactions between polyphenols
and polysaccharides: mechanisms and consequences in food processing and diges-
tion, Trends Food Sci. Technol. 60 (1) (2017) 43–51.
[77] J.H. Waite, N.H. Andersen, S. Jewhurst, C. Sun, Mussel adhesion: finding the tricks
worth mimicking, J. Adhes. 81 (3–4) (2005) 297–317.
[78] J. Yang, M.A. Cohen Stuart, M. Kamperman, Jack of all trades: versatile catechol
crosslinking mechanisms, Chem. Soc. Rev. 43 (24) (2014) 8271–8298.
[79] J. Yang, V. Saggiomo, A.H. Velders, M.A. Cohen Stuart, M. Kamperman, Reaction
pathways in catechol/primary amine mixtures: a window on crosslinking chemis-
try, PLoS One 11 (12) (2016) e0166490.
[80] L.Q. Xu, D. Pranantyo, J.B. Liu, K.-G. Neoh, E.-T. Kang, Y.X. Ng, S. Lay-Ming Teo, G.D.
Fu, Layer-by-layer deposition of antifouling coatings on stainless steel via
catechol-amine reaction, RSC Adv. 4 (61) (2014) 32335–32344.
[81] L. Shen, H.-F. Ji, The pharmacology of curcumin: is it the degradation products?
Trends Mol. Med. 18 (3) (2012) 138–144.