International Journal of Biological Macromolecules xxx (xxxx) 1–10

Contents lists available at ScienceDirect

International Journal of Biological Macromolecules

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

F
OO
Curcumin loaded waste biomass resourced cellulosic nanofiber cloth as a
potential scaffold for regenerative medicine: An in-vitro assessment
a b a b a b b
Nurul Nadirah Suteris , , Amina Yasin , , Izan Izwan Misnon , , Rasidi Roslan ,
b b b ⁎ a b ⁎
Farah Hanani Zulkifli , Mohd Hasbi Ab Rahim , Jayarama Reddy Venugopal , , Rajan Jose , ,

PR
a Center for Advanced Intelligent Materials, Universiti Malaysia Pahang, 26300 Kuantan, Malaysia
b Faculty of Industrial Sciences and Technology, Universiti Malaysia Pahang, 26300, Kuantan, Malaysia

ARTICLE INFO ABSTRACT

Keywords: This article demonstrates the development of nanofibrous cloths by electrospinning of renewable materials, i.e.,
Natural medicine
Renewable materials
Skin tissue engineering
Surface wetting
Adsorbent polymers
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curcumin-loaded 90% cellulose acetate (CA)/10% poly(ε-caprolactone) (PCL), for applications in regenerative
medicine. The CA is derived from the biomass waste of the oil palm plantation (empty fruit bunch). The
nanofiber scaffolds are characterized for the fiber morphology, microstructure, thermal properties, and wettabil-
ity. The optimized smooth and bead-free electrospun fiber cloth contains 90% CA and 10% PCL in two curcumin
compositions (0.5 and 1 wt%). The role of curcumin is shown to be two-fold: the first is its function as a drug and
the second is its role in lowering the water contact angle and increasing the hydrophilicity. The hydrophilicity
enhancements are related to the hydrogen bonding between the components. The enhanced hydrophilicity con-
CT
tributed to improve the swelling behavior of the scaffolds; the CA/PCL/Cur (0.5%) and the CA/PCL/Cur (1.0%)
showed swelling of ~700 and 950%, respectively, in phosphate-buffered saline (PBS). The drug-release studies
revealed the highest cumulative drug release of 60% and 78% for CA/PCL/Cur (0.5%) and CA/PCL/Cur (1.0%)
nanofibers, respectively. The in-vitro studies showed that CA/PCL/Cur (0.5 wt%) and CA/PCL/Cur (1.0 wt%)
nanofiber scaffolds facilitate a higher proliferation and expression of actin in fibroblasts than those scaffolds
without curcumin for wound healing applications.
RE

1. Introduction many applications such as filtration, cosmetic, pharmaceutical, veteri-

Development of functional materials from renewable sources have
become one of the top priorities in the modern world not only due to
the depletion of natural resources but also to eliminate the adversities
OR
nary food and tissue engineering [8–10].
Recently, in regenerative medicine, wound dressings based on many
natural polymers such as polylactic acid (PLA), gelatin, fibrin, keratin,
chitosan, silk fibroin etc., mainly extracted from plant and animal

due to their over usages such as piling up of non-degradable used prod-
ucts and increased atmospheric content of carbon [1–3]. One of the
most desirable remedies to tackle this issue is using renewable materi-
als without affecting food security. Cellulose, which is a polysaccharide
found in plants and living organisms [4], holds the forerunner's baton
because of the resource enormity. Widening the application domains of
cellulose have become a top priority in the sustainable materials com-
C
sources including cellulose have attracted great attention compared to
the traditional methods of graft or allogenic skin tissue [11–12]. Among
these natural polymers, cellulose is particularly attractive due to its
unique properties that can mimic the characteristics of extracellular
matrices (ECM) [13] along with its resource sustainability. Different
types of wound dressings materials such as electrospun nanofiber scaf-
folds, foams, hydrocolloids and hydrogels are available in the open

munity because many industrial crops result in a large number of by- market [14–15]. Among them, electrospun nanofiber scaffolds have ex-
products besides their primary target, most of them are sources of cellu- clusive advantages [16] such as high porosity to absorb the excess
lose. For example, in the case of the oil palm sector, statistics show that wound exudates and to inhibit penetration of microorganism, high spe-
UN

~99 kg of by-products are generated for each one-kilogram crude palm cific surface area for drug loading and delivery, oxygen penetration
oil, ~20% of this by-product is cellulose-rich empty fruit bunch (EFB) through the dressing and reach to the wound site, water vapor transmis-
[5–7]. The cellulose extracted from EFB could be further developed for sion to provide required moisture for the wound healing, excellent per-

⁎ Corresponding authors.
E-mail addresses: venugopal@ump.edu.my (J.R. Venugopal), rjose@ump.edu.my (R. Jose).

https://doi.org/10.1016/j.ijbiomac.2021.12.006
Received 17 May 2021; Received in revised form 11 November 2021; Accepted 1 December 2021
0141-8130/© 2021

Note: Low-resolution images were used to create this PDF. The original images will be used in the final composition.
N.N. Suteris et al.
formances in cell adhesion, proliferation, migration, and differentiation
as well as physical properties similar to that of ECM [17–19]. Besides,
electrospun bio-scaffolds hold unique properties such as high biocom-
patibility, offer enough protection to any exposure to the wound site
against bacteria and dust, promote epithelization, and provide enough
pores for the exchange of gases [20–23]. Despite these exciting advan-
tages, many of these scaffolds have drawbacks such as scar formation,
contraction of wounds, and weak integration with host cell/tissue. To-
wards this end, three-dimensional (3D) scaffolds have attracted more
attention because they can cover the wounds and create a strong physi-
cal barrier against external infection [24]. The 3D porous networks play
an important role in guiding as a new template for the formation of skin
tissues [25–26].
Although cellulose offers advantages as a wound healing medium,
electrospun cellulose nanofibers scaffolds are limited because of the dif-
ficulties in choosing affordable and environmental friendly solvents.
Besides, the healing rate of the cellulose scaffolds is limited due to the
poor efficiency to absorb exudate and avoid bacterial growth [27]. In
contrast, cellulose acetate (CA), a derivative of cellulose having struc-
tural and compositional similarity to the glycosaminoglycan (compo-
nents of the ECM), has attracted numerous studies due to its biocompat-
ibility, high surface tension, hydrophilicity, structural integrity,
biodegradability, and processability into ultrafine nanofibers [28–29].
However, CA has few disadvantages, such as the lack of proper active
healing capability, low breaking stress, poor resistance and reduced an-
tibacterial activity [30–31]. The functions of CA can be improved by
blending with suitable polymers such as polylactic acid (PLA), gelatin,
polyurethane (PU), polyvinyl alcohol (PVA), and poly(ε-caprolactone)
(PCL) [28,30,32]. Among them, PCL was used for wound healing appli-
ED
cations due to its excellent biodegradation, high biocompatibility, ade-
quate tensile strength, as well as FDA approval [33,34]. The CA and the
CT
PCL nanofibers could help to introduce an appropriate approach for ob-
taining a matrix with both properties of bioactivity affinity and good
mechanical stability. The CA nanofibers are incorporated with a wide
range of therapeutic agents ranging from antibacterial, anti-
inflammatory, antiviral and antioxidant activity [15,35]. For example,
wound healing drugs, such as curcumin (natural) or Neosporin (syn-
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thetic), can be dispersed along with CA and could be developed into fi-
brous scaffolds for regenerative tissue engineering [36–37]. In addi-
tion, the incorporation of therapeutic agents like aloe vera, manuka
honey, zataria multiflora, berberine, Malva sylvestris etc., through CA/
PCL polymeric blend matrix could help in raising the potential applica-
tions. Among all, curcumin (1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-
heptadiene3,5-dion) (Cur), a component in turmeric isolated from the
OR
family of Curcumin longa [38], plays an important role in addressing
the issue of wound healing in the biomedical field [39–40]. The past
decade has shown a rapid development of curcumin in various research
in terms of its biological applications and its unique molecular structure
for wound healing applications [41–42]. Curcumin exhibits a strong an-
tioxidant, anti-inflammatory, anti-bacterial, anti-virus, anti-tumour
and anti-infective properties [20,40,42–43].
C
This study is to synthesize and characterize the electrospun CA/
PCL/Curcumin cloths as a drug-loaded wound healing membrane. The
optimized composition for getting smooth and bead-free electrospun
UN
fiber cloth contains 90% CA and 10% PCL in two compositions of cur-
cumin (0.5 and 1 wt%). The curcumin has played two critical roles, the
first is its function as a drug and the second is reducing the water con-
tact angle for superior hydrophilicity. The origin of this hydrophilicity
is identified and discussed. To the best of our knowledge, the fabrica-
tion of CA/PCL/Cur nanofibers scaffold by using CA obtained from ex-
tracted cellulose of EFB of palm plantation is reported here for the first
time. To date, there are not many ideal skin scaffolds that have been
successfully fabricated to be used in tissue engineering despite much re-
search dedicated to fabricating these biomaterials. The present study
develops electrospun CA/PCL/Cur nanofibers that could help deepen
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International Journal of Biological Macromolecules xxx (xxxx) 1–10
the future understanding and knowledge to improve the performance of
skin scaffolds for tissue engineering.
2. Materials and methods
2.1. Materials
F
Cellulose acetate was sourced from the cellulose extracted from
empty fruit bunch (EFB) of Palm plantations. Poly(ε-caprolactone)
OO
(PCL) with a molecular weight of 80,000 g/mol, curcumin (Cur; yellow
powder) and phosphate buffer saline were purchased from Sigma
Aldrich, United State. Glutaraldehyde (GA) solution (30% aqueous so-
lution) and acetone were purchased from Merck, Germany. Methanol
and chloroform were purchased from Fisher Chemical, United King-
dom. Glacial acetic acid was purchased from Friendmann Schmidth,
Australia. Dulbecco's Modified Eagle Medium (DMEM) was purchased
from Life Technologies, USA. Actin (ThermoFisher Scientific, Singa-
PR
pore), sodium chlorite (NaClO2), dichloromethane (CH2Cl2) and
sodium hydroxide (NaOH) were purchased from Merck, Germany. The
3T3 mouse fibroblast cells were purchased from ATCC, USA. All the
chemicals and materials were used without further purification. All so-
lutions were prepared using Millipore water.
2.2. Isolation of cellulose from EFB of palm plantation
Cellulose was isolated from EFB of palm plantation by slight modifi-
cation of the previously reported method (Fig. S1, Supporting Informa-
tion) [44]. The EFB was first pretreated by hexane–methanol (2:1, v/v)
for 5 h to remove oil/wax from the sample, followed by a two-step
process. Firstly, the cleaned EFB was treated with 5 wt% NaOH solution
at a solvent/material ratio of 30:1 at 80 °C for 2 h under mechanical
stirring. The slurry was cooled, filtered, and washed with distilled wa-
ter until neutral. The residue was then bleached with 1.7% w/v sodium
chlorite (NaClO2) at 30:1 ratio of solvent/material, acidified the solu-
tion with 80% acetic acid to reach pH ~4 for delignification and
washed with distilled water. The bleaching was repeated twice at 70 °C
for 4 h. After bleaching, the solution was filtered and the residue was
washed with distilled water several times to remove the yellowish color
and odor to get neutral and colorless material and then oven-dried at
60 °C until constant weight. The extracted cellulose was characterized
by 1HNMR, FTIR, XRD and TGA (Fig. S3). The extracted cellulose was
then further used for the synthesis of cellulose acetate.
1HNMR (CD Cl, 500 MHz): δ 4.22 (s, 2H), 4.12 (d, J = 5 Hz, 1H),
3
4.02 (d, J = 12.5 Hz, 2H), 3.97 (s, J, 1H), 3.95 (d, J = 10 Hz, 1H), 3.82
(br s, 1H), 3.73 (d, J = 10 Hz, 1H), 3.31 (d, J = 5 Hz, 1H). IR: 3345 (ѵ
O H), 2904 (ѵ -C-H), 2849 (ѵ -C-H), 1647 (ѵ -OH), 1430 (δ -
CH2),1163 (ѵ -C-O).
2.3. Acetylation of cellulose
To convert the cellulose into CA, the green acetylation of cellulose
was carried out based on the previous method [45] with slight modifi-
cation. A mixture of 0.4 g of extracted cellulose, 20 mL acetic anhy-
dride and 0.6 g of iodine was stirred and heated in 100 mL round-
bottomed flask at 80 °C for 5 h. After cooling the mixture to ambient
temperature, 10 mL saturated sodium thiosulfate solution was added
and stirred until the solution became colorless due to the reduction of
iodine to iodide. The solution was then transferred into a beaker con-
taining 60 mL of ethanol at 0 °C and stirred for 60 min. The resulting
solution was filtered and washed thoroughly with ethanol (75% v/v)
and distilled water to remove the unreacted acetic acid and other by-
products. The solid residue was dried at 60 °C in a vacuum oven, dis-
solved in dichloromethane (DCM), filtered it, and then concentrated the
filtrate under vacuum. Cellulose acetate was formed as a film, which
N.N. Suteris et al.
was further dried at 60 °C in a vacuum oven for 24 h. The CA was char-
acterized by 1HNMR, FTIR, XRD and TGA (Fig. S3).
1HNMR (CD Cl, 500 MHz): δ 5.07 (t, J = 10 Hz, 2H), 4.80 (t, J = 5 &
3
10 Hz, 1H), 4.43 (br d, 2H), 4.07 (br s, J, 1H), 3.73 (d, J = 10 Hz, 1H),
3.58 (s, 1H), 2.09 (s, 3H), 2.01 (s, 3H), 1.95 (s, 3H). IR: 2958 (ѵ -C-H),
2881 (ѵ -C-H), 1736 (ѵ -C=O), 1372 (δ -CH3), 1235 (ѵ -C-O).
2.4. Preparation of polymeric solutions for electrospinning
Procedure adopted for the synthesis of the nanofiber cloths is
schematically displayed in Fig. 1. Different weight ratio (10% w/v, 13%
w/v and 15% w/v) of CA and PCL solutions (10% w/v, 13% w/v and
15% w/v) were prepared in order to get smooth, homogenous, and vis-
cous solutions suitable for electrospinning. The best weight ratio was
identified as 13% w/v and 15% w/v for CA and PCL, respectively, for
the bead-free nanofiber formations. The 13% by weight of CA spinning
solution was prepared by dissolving 9 g of CA in 100 mL of 3:1 acetic
acid/acetone mixture. Similarly, 1.5 g of PCL was dissolved in a 100 mL
solution of 3:1 chloroform/methanol mixture to obtain 15% by weight
of PCL. The CA/PCL blend solution was prepared by adding 90 mL of a
clear viscous CA solution (13%) into 10 mL of PCL (15%) solution to get
90:10 CA/PCL solution. For making the CA/PCL/Cur nanofibers, appro-
priate amounts of curcumin were added to the above CA/PCL formula-
tion in 0.5% and 1.0% by weight (with respect to CA/PCL blend). All
the solutions were stirred overnight on a magnetic stirrer at 1000 rpm
at room temperature in closed containers to ensure homogeneity; the
solutions were ultrasonically degassed for 5 min before electrospin-
ning.
ED
2.5. Electrospinning
CT
The CA/PCL/Cur solutions were electrospun into nanofiber mats us-
ing a pilot scale electrospinning machine employing 100 needles
(Bioinicia, Fluidnatek, Spain). Approximately 20 mL of the uniformly
mixed solution was fed into a plastic syringe fixed with a 20-gauge nee-
dle of diameter ~0.9 mm and electrospun at 15 kV with a solution feed
rate of 1.0 mL/h at room temperature (27 °C) and relative humidity of
RE
~50%. The distance between the tip of the needles and the collector
was adjusted to be ~10 cm. The resulting nanofiber scaffolds were col-
lected from the aluminium foil, vacuum dried at 40 °C for 24 h to elimi-
nate the solvent residue, and then stored in a desiccator for later use.
2.6. Structural characterization
OR
The surface morphology of the electrospun nanofibers mats was
studied by field emission scanning electron microscopy (FESEM, JSM-
7800F, JEOL) at magnifications of 5000× and 40,000×. The func-
tional groups in the nanofiber mats were studied by attenuated total re-
flectance – Fourier transform infrared (ATR-FTIR) spectrometer (Spec-
trum One, Perkin-Elmer, USA) over a range of 700–4000 cm−1 at a reso-
C
UN
Fig. 1. Flowchart and electrospinning parameters for the synthesis of pure CA,
PCL, CA/PCL blend and CA/PCL/Cur nanofiber scaffolds.
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International Journal of Biological Macromolecules xxx (xxxx) 1–10
lution of 4 cm−1. The crystallinity and crystal structure of the
nanofibers were studied using X-ray diffraction technique (Rotaflex
RT300 mA, Rigaku Corporation, Osaka, Japan) at a scanning speed of
2θ = 1°/min. Proton nuclear magnetic resonance (1H NMR) spectra of
pure Cur, CA, PCL, CA/PCL, CA/PCL/Cur nanofiber scaffolds and
loaded Cur from CA/PCL/Cur scaffolds were recorded on a Bruker ul-
tra-shield plus 500 (500 MHz) using tetramethyl silane (TMS) as an in-
F
ternal standard. The coupling constant was calculated in Hz and chemi-
cal shifts (δ) in ppm. Multiplicities of peaks are denoted as: s = singlet,
OO
d = doublet, dd = double doublet, t = triplet and m = multiplet. The
thermal behavior of the nanofiber scaffolds was determined by thermo-
gravimetric analysis (TGA, Mettler Toledo TGA/DSC-1600) in the
20–450 °C range in nitrogen atmosphere at a rate of 10 °C/min.
2.7. Characterization of nanofibers as wound healing mats
The surface wettability of the nanofiber scaffolds was examined by
PR
Data Physics Optical Contact Angle with SCA-20 system. Sessile droplet
of Phosphate-buffered saline (PBS) was poured at the center of each
nanofiber scaffold and the interaction between the PBS and the surface
was measured through the video recorder of the instrument. The
swelling and the degradation of scaffolds were studied using nanofibers
mats of 10 mm × 10 mm soaked in 8 mL of PBS solution (pH ~7.2) at
27 °C. The samples were analysed for every 3, 6 and 9 days. The experi-
mental details of the degradation study are in the Supporting informa-
tion. The in-vitro Cur release was analysed by immersing each CA/PCL/
Cur nanofiber scaffold in PBS solutions (37 °C, pH ~7.4) as a release
medium. Approximately 6 mg of CA/PCL/Cur nanofiber scaffolds was
immersed in 10 mL of PBS solution and incubated at 37 °C for varying
time periods (24, 48, 72 and 120 h). At regular intervals, 1 mL of PBS
aliquot was examined using UV–Vis absorption spectra (UV–Vis-NIR
spectrophotometer UV-2600 SHIMADZU, Japan) at λmax = 421 nm, at
which wavelength the absorption maximum of curcumin occurs. The
percentage of drug release was determined using the absorption data of
curcumin. The standard MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-
diphenyltetrazolium bromide) assay was carried out to assess cytocom-
patibility of biomaterials scaffolds by colorimetric measurement of for-
mazan formed by metabolically active fibroblast cells on the
nanofibers. For the test, the CA/PCL/Cur scaffolds were cultured on
3T3 fibroblasts. The cell viability was evaluated using a microplate
reader (TECAN SAPPHIRE). The results were evaluated by microscope
observation during the 3rd, 6th, and 9th day at 10× magnification. The
3T3 mouse fibroblast cells were stained with a fluorescent dye 4-(4-
(chloromethyl)benzamido)-2-(1,2,2,4,8,10,10,11-octamethyl-
1,2,10,11-tetrahydropyrano [3,2-g:5,6-g']diquinolin-13-ium-6-yl)
benzoate (CMTPX) to monitor the movement of the live cells. After 3, 6
or 9 days of cell growth, the complete cell culture medium was safely
removed from the scaffolds seeding 24-well plates and cell were fed
with DMEM medium. The CMTPX dye was then added to the cells-
scaffolds and further incubated at 37 °C for 3 h. Thereafter, the CMTPX
dye medium was exchanged by complete medium, and cells were incu-
bated at 37 °C for 24 h. After one day incubation, the medium was re-
moved, and the biomaterials scaffolds were rinsed once again with PBS
to remove dead cells. The scaffolds were then viewed under the in-
verted microscope after being added with a serum-free medium. For
DAPI staining, the samples were incubated with tetramethyl rhodamine
isothiocyanate (TRITC)-conjugated phalloidin in the dilution of 1:200
for 90 min at room temperature. The treated samples were washed with
PBS thrice to remove the excess staining and then incubated with DAPI
in the dilution of 1:3000 for 30 min at room temperature. The samples
were then removed and mounted over a glass slide using Vectashield
mounting medium and examined under Olympus FV1000 (USA) fluo-
rescent microscope. For morphology analysis, the fibroblast cells grown
on scaffolds were rinsed twice with PBS and fixed in 3% glutaraldehyde
for 30 min. Thereafter, the scaffolds were dehydrated with increasing
N.N. Suteris et al.
concentrations of alcohol (20, 40, 60, 80 and 100%). The samples were
air dried by keeping the samples in a fume hood. Lastly, the scaffolds
were observed using SEM at an accelerating voltage of 10 kV (FESEM,
JSM-7800F, JEOL). The seeding of the nanofibers and MTT assay are
detailed in Supporting information.
2.8. Statistics
The data were obtained from six samples: their mean and standard
deviation (SD) were determined and reported in this article.
3. Results and discussion
The CA/PCL polymer blends at different volumetric ratios were suc-
cessfully synthesised following an optimization process as detailed in
the Supporting Information (Table S1). Fig. 2(a–c) shows the color of
the spun nanofiber mats, morphology of the fibers constituting the
mats, and the statistics of the diameter distribution of nanofibers of CA/
PCL scaffolds with and without Cur. Similar details of CA and PCL are
shown in the Supporting Information (Fig. S4(b)). The presence and
concentration of curcumin in the CA/PCL scaffold mats was evident
through a color change (Fig. 2a–c (i)). All the CA/PCL/Cur samples
showed typical random fiber web of bead-free electrospun morphology
(Fig. 2a–c (ii & iii)); the optimum quantity of curcumin in 90/10 CA/
PCL was ~1% above which bead-free nanofibers could not be devel-
oped (Fig. S4(c)). Fig. 2a–c(iv) show the statistics of nanofiber diame-
ters measured from 150 fibers. The average diameter for PCL and CA
nanofibers mats was ~860 and ~430 nm, respectively (Fig. S4(d)). The
CA/PCL blend nanofibers mats showed further lowering of nanofiber
ED
diameter (~137 nm). The addition of curcumin into CA/PCL resulted in
an increase in the average diameter of ~148 nm and ~152 nm for Cur
CT
0.5 wt% and Cur 1.0 wt% respectively (with respect to CA/PCL blend).
The diameter of the nanofiber scaffolds was increased by the cumula-
tive curcumin content in the scaffolds. These diameters support the cell
attachment because of the required diameters of nanofiber scaffolds
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C OR
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Fig. 2. (i) Photographs of the electrospun nanofibers containing CA/PCL and CA/PCL/Cur nanofiber scaffolds. (ii) FESEM images of electrospun nanofibrous scaf-
folds at 5000, (iii) FESEM images of electrospun nanofibrous scaffolds at 40000, (iv) diameter distribution histograms of (a) CA/PCL blend; (b) CA/PCL/Cur
(0.5 wt%) and (c) CA/PCL/Cur (1.0 wt%).
4
International Journal of Biological Macromolecules xxx (xxxx) 1–10
and ECM for cell attachment and proliferation with the diameter of fi-
broblast cells ranging from 1 to 4 μm [13,46].
The thermal stability of the CA/PCL/Cur nanofibers was studied by
TGA (Supporting Information, Fig. S5). A weight loss of ~2–4% was ob-
served in the 20–200 °C temperature range for all the samples due to
evaporation of adsorbed water in the nanofibers as shown in Fig. S5.
Next, a major weight loss of ~80% was observed in the 200–450 °C
F
temperature region corresponding to the degradation of C O, C H
and O H side chains. Degradation temperature of cellulose is reported
OO
to be in the range 300–450 °C, depending on the additive [47]. Rela-
tively higher complete decomposition temperature (~450 °C) in the
present case could be attributed to the hydrogen bonding between the
CA/PCL and Cur as well as due to the amorphization as observed in the
XRD patterns (Fig. 3b). This change could be beneficial to enhance the
drug dissolution in the polymer matrix [48–49].
One of the important aspects of a scaffold for regenerative medicine
is the chemical structure of the materials. Chemical contents of the
PR
fibers were studied using FTIR (Fig. 3a), XRD (Fig. 3b), and NMR (Fig.
3c). The FTIR spectra of pure CA, PCL and Cur spectra are compared to
their composites in Fig. 3a for analysing the functional groups as well as
to examine whether any chemical reaction has happened during mixing
them into a single solution and by exposing to a high electric field re-
quired for electrospinning. The FTIR spectrum of CA/PCL is dominated
by the vibrations of CA most likely because of its higher content; how-
ever, characteristic peaks of PCL are also present as evident from the in-
crease in the peak intensity at the 2938–2865 cm−1 range. Besides, in
the fingerprint region (1470–1100 cm−1), the appearance of a combina-
tion of the peaks from both CA and PCL and changes in their relative in-
tensity have become prominent, indicating the incorporation of both
polymers. The FTIR spectra of the Cur containing scaffolds (Fig. 3a(v &
vi)), exhibit the features consisting of characteristic bands of both pure
Cur and CA. Notably, the sharp phenolic −OH peak of Cur is disap-
peared at 3477 cm−1, however, the occurrence of other characteristic
peaks of Cur such as carbonyl peak with a slight shift at 1623 cm−1 for ѵ
(C=O), aromatic peaks at 1551 cm−1 and 1514 cm−1 for symmetric
ѵ(C=Cring) and δ(C H) in-plane bending at 1216 cm−1 of the aro-
N.N. Suteris et al.
ED
Fig. 3. Characterization of the nanofibers. (a) FTIR spectra of pure Cur, pure
CA nanofibers, PCL nanofibers, CA/PCL blend, CA/PCL/Cur (0.5 wt%) and CA/
CT
PCL/Cur (1.0 wt%). (b) XRD diffractogram of pure Cur, pure CA nanofibers,
PCL nanofibers, CA/PCL blend, CA/PCL/Cur (0.5 wt%) and CA/PCL/Cur
(1.0 wt%). (c) NMR spectra of CA/PCL/Cur nanofibers. (d) NMR spectra of (i)
pure Cur and (ii) loaded Cur obtained by dissolution of CA/PCL/Cur
nanofibers.
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matic rings confirm the successful incorporation of Cur with polymers.
The characteristic peak at 1366 cm−1 of δ(-CH3) bending vibration for
CA with a slight shift attributes the presence of CA in Cur-incorporated
nanofibers mats. Shifting of peaks towards lower wavenumbers and
changes in their relative intensities of the 1740–1200 cm−1 bands could
be the indication of the interactions between Cur functional groups
with hydroxyl, carbonyl and ether groups of the polymers which is sup-
OR
posed to be occurred due to the intermolecular hydrogen bonding be-
tween Cur and polymers. The hydrogen bonds in nanofibers could pro-
mote the hydrophilicity of the scaffolds; this enhancement is highly
beneficial for the biological environment. Therefore, the addition of
Cur through CA/PCL blend could contribute to the progression of bioac-
tivity.
To further confirm the above structural assignments, the NMR of the
C
Cur containing samples were recorded and compared with their coun-
terparts (Fig. 3c). The NMR chemical shift (ppm) of composite CA/PCL
and CA/PCL/Cur nanofibers could be noticed in the same region as that
UN
for pure CA, PCL, and Cur (Fig. S6) without the addition of any new sig-
nal, thereby indicating that the Cur incorporation may occur due to in-
termolecular hydrogen bonding interaction between functional groups
of Cur and the polymers rather than any chemical modification. The
characteristic peaks of CA at δ5.2–3.5 (ppm) and that of PCL as two
triplet peaks at δ4.09–4.07 (ppm) and δ2.34–2.31 (ppm) could be ob-
served in 1HNMR spectra of both CA/PCL (Fig. S6(c)) and CA/PCL/Cur
nanofibers scaffold (Fig. 3d), an indication of the presence of both poly-
mers in nanofibers scaffolds. However, the additional Cur signals at
aromatic region (δ7.60–5.88 ppm) with very slight chemical shift could
easily be noticed in 1HNMR spectra of CA/PCL/Cur scaffold (δ: 9.96 (s,
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International Journal of Biological Macromolecules xxx (xxxx) 1–10
2H, Cur-OH), 7.63 (d, 2H, Cur), 7.16 (dd, 2H, Cur), 7.08 (d, 2H, Cur),
6.97 (d, 2H, Cur), 6.52 (d, 2H, Cur), 5.88 (s, 2H, Cur) 5.14 (t, 1H, CA),
4.82 (br s, 1H, CA), 4.44 (br d, 2H, CA) 4.08 (t, 2H, PCL), 3.97 (s, 6H,
Cur-methoxy), 3.74 (br s, 1H, CA), 3.60 (d, 1H, CA), 2.33 (t, 2H, PCL),
2.15–1.97 (t, 9H, CA-acetyl), 1.70–1.63 (m, 4H, PCL) & 1.42–1.37 (m,
2H, PCL)) with same characteristic as that of pure Cur (δ: 7.61 (d, 2H),
7.13 (dd, 2H), 7.05 (d, 2H), 6.94 (d, 2H), 6.49 (d, 2H), 5.86 (s, 2H) &
F
3.95 (s, 6H)) which support the successful incorporation of Cur with
CA/PCL nanofibers. Moreover, there is no additional peak, demonstrat-
OO
ing that the nanofiber scaffolds do not induce any chemical change and
support the physical interaction (intermolecular hydrogen bonding) oc-
curred between Cur and polymers. The presence of an additional peak
at 9.96 ppm is due to the OH proton signal of Cur, present only in
1HNMR spectra of CA/PCL/Cur nanofibers, supporting the strong evi-
dence of intermolecular interaction by hydrogen bonding. The disap-
pearance of the OH proton signal in the 1HNMR spectrum of pure Cur is
due to the exchange of OH protons with the deuterium of deuterated
PR
solvent. However, this effect is reduced by intermolecular hydrogen
bonding between Cur and the polymers and due to which the OH pro-
ton signal could be easily noticed in 1HNMR spectra of CA/PCL/Cur
nanofibers. In order to check the chemical integrity of incorporated
Cur, 1H NMR spectra of Cur released from nanofibers are also compared
with that of pure Cur. It can be seen from Fig. 3d (i & ii), loaded Cur, ob-
tained by dissolution of CA/PCL/Cur nanofiber scaffold, exhibit the
aromatic proton signals at 7.60–5.89 ppm and OCH3 proton signal at δ
3.9 ppm similar to that of pure Cur, attributing the chemical integrity
to be retained in the Cur incorporated nanofibers.
The changes in crystallinity of the Cur containing samples were
studied by XRD (Fig. 3b). The pure Cur showed sharp and intense peaks
at 2θ ~7.96, ~8.78, ~12.26, ~14.54 and ~17.16 representing the
(110), (101), (111), (211), (220), and (112) planes and various lower
intensity peaks at 2θ ~ 30–44°, indicating that curcumin is highly crys-
talline [50]. Similarly, PCL also showed an intense sharp peak at 2θ
~21° and a relatively lower intensity peak at 2θ ~23.5° corresponding
to the (110) and (200) plane reflections, respectively, of a polyethylene-
like crystal structure [51–52]. On the contrary, CA shows an amor-
phous structure with two broad peaks at 2θ ~10° and ~19°. Addition of
impurity in a polymer has been reported to result in deterioration of
crystalline structure as the incorporated impurity causes misalignment
in the polymer chains [53]. The XRD pattern of CA/PCL nanofibers
(Fig. 3b) showed decrease in intensity of the peaks in both polymers, in-
dicating the deterioration in the crystalline structure of PCL. In the case
of XRD of CA/PCL/Cur nanofibers, the characteristic peaks of crys-
talline Cur completely disappeared indicating that the crystalline Cur
existed as an amorphous nano solid dispersion after electrospinning.
This observation supports the molecular level dispersion of Cur in the
CA/PCL/Cur nanofibers and also reveals that Cur was well blended in
the CA/PCL nanofiber because of intermolecular hydrogen bonding be-
tween Cur and different functionalities present in the polymers.
Results of static water contact angle (Φ) measurements and surface
wettability of the CA/PCL/Cur nanofibers scaffolds are shown in Fig. 4
(a). The measured Φ followed the order of PCL > CA > CA/
PCL > CA/PCL/Cur (0.5%) > CA/PCL/Cur (1.0%), indicating that the
CA/PCL/Cur nanofiber scaffolds are preferable to be used for cells at-
tachment and proliferation. The obtained Φ of the PCL and the CA
nanofiber are 128° ± 7 and 36° ± 4, respectively, showing hydropho-
bicity of the PCL whereas hydrophilicity of CA nanofibers. Interest-
ingly, the Φ decreased further to 33° ± 2 as in CA/PCL blend nanofiber
mats, indicating that PCL could increase the surface wettability of CA
nanofibers. This increase in hydrophilicity of CA/PCL blend nanofibers
could be explained as due to intermolecular interaction (hydrogen
bonding) of polymers, as shown in Scheme 1. Further addition of cur-
cumin reduced the Φ to ~25° ± 1 and ~19° ± 2 for CA/PCL/Cur
(0.5%) and CA/PCL/Cur (1.0%), respectively, owing to the increased
hydrogen bond as demonstrated in Scheme 1. Therefore, adding Cur to
N.N. Suteris et al. International Journal of Biological Macromolecules xxx (xxxx) 1–10

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PR
ED
CT
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Fig. 4. (a) Water contact angle measurement of pure CA, PCL, CA/PCL blend, CA/PCL/Cur (0.5 wt%) and CA/PCL/Cur (1.0 wt%) nanofibers. (b) Weight loss of
CA, PCL, CA/PCL blend, CA/PCL/Cur (0.5 wt%) and CA/PCL/Cur (1.0 wt%) nanofibers scaffolds after degradation in PBS medium for up to 9 days. (c) Swelling
properties of the biocomposite nanofibers scaffolds. Data's are presented as mean ± standard deviation with n = 6 (** P˂0.01, * P ˂ 0.001). (d) The drug release
of CA/PCL/Cur nanofiber scaffolds for 0.5% and 1.0% curcumin. (e) Proliferation of fibroblasts on functionalized biocomposite nanofibrous scaffolds. Data's are
presented as mean ± standard deviation with n = 6 (** P˂0.01, * P ˂ 0.001). (f) CMTPX live cell imaging of fibroblast cells on biocomposite nanofibrous scaf-
folds on day 5 (i) control, (ii) CA/PCL, (iii) CA/PCL/Cur 0.5 wt%, and (iv) CA/PCL/Cur 1.0 wt%.
OR

the CA/PCL scaffolds further enhances wound exudate absorption (as
detailed subsequently) by maintaining the moisture on the wound side
in addition to its medicinal properties. The increase in hydrophilicity of
CA/PCL/Cur nanofibers could also help in cell proliferation and migra-
tion on nanofibers.
Scheme 1 shows that the formation of composite CA/PCL/Cur scaf-
C
4a) strongly support the formation of CA/PCL/Cur scaffolds by in-
tramolecular & intermolecular hydrogen bonding by increasing the hy-
drophilicity of scaffolds as compared to their characteristic com-
pounds. Therefore, all the structural analysis indicates that no chemi-
cal modification or chemical reaction occurred due mixing of polymers
with and without Cur and exposing this solution to a high electric field

folds is most likely to be occurred by intermolecular hydrogen bonding during electrospinning.

interaction between the molecules. The synthesis of nanofibers is
mainly by two ways i.e., (i) chemical interaction by forming the new 3.1. Scaffold degradation analysis
bonds which usually show the different or additional evidence in struc-
UN

tural analysis and (ii) physical interactions such as hydrogen bonding
and electrostatic interaction. However, here it is closely observed by
the evidence obtained from FTIR (Fig. 3a), XRD (Fig. 3b) and NMR
(Fig. 3c) that the CA/PCL/Cur scaffolds formation is by intramolecular
and intermolecular hydrogen bonding rather than chemical interac-
tion. There is no evidence such as the presence of any additional new
peak or signals in FTIR and NMR could be observed which support the
structural modification of the components. The hydroxyl and carbonyl
groups of Cur may have the possibility of intermolecular hydrogen
bonding with the carbonyl group of PCL and hydroxyl, carbonyl, and
ester groups of CA. Moreover, the contact angle measurements (Fig.
The weight loss of the nanofiber samples during the in-vitro degra-
dation is shown in Fig. 4b. The results indicate that the fabricated
nanofibers undergo significant degradation (weight loss) up to 9 days
and remain structurally stable after that. The in-vitro degradation of the
nanofiber scaffolds followed the order of CA > CA/PCL/Cur
(1.0%) > CA/PCL/Cur (0.5%) > CA/PCL > PCL, indicating the
slower degradation rate (4%) in PCL as compared to other nanofibers
mainly because of its hydrophobic behavior. The hydrophobicity of PCL
also influences the degradation rate of CA by lowering its weight loss
from 38 to 34% in CA/PCL nanofibers. In general, the degradation of
the electrospun nanofiber scaffolds is influenced by various factors, in-

6
N.N. Suteris et al. International Journal of Biological Macromolecules xxx (xxxx) 1–10

F
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PR
ED
Scheme 1. The formation of CA/PCL/Cur scaffolds is proposed by intramolecular & intermolecular hydrogen bonding. The possibility of hydrogen bonding in the
composite CA/PCL/Cur scaffolds formation include (i) intermolecular hydrogen bonding between hydroxyl group of Cur and carbonyl group of CA, (ii) Cur as exist
in keto–enol tautomers may have intramolecular hydrogen bonding, (iii) intermolecular hydrogen bonding between the hydroxyl groups of Cur molecules, (iv) in-
termolecular hydrogen bonding of enol form of Cur with the carbonyl group of PCL, (v) intermolecular hydrogen bonding of hydroxyl group and ester group of CA,
(vi) intermolecular hydrogen bonding of hydroxyl group and carbonyl group of CA, (vii) hydrogen bonding between hydroxyl and carbonyl group of Cur, (viii) in-
termolecular hydrogen bonding between hydroxyl group of CA and carbonyl group of PCL, and (ix) intermolecular hydrogen bonding of hydroxyl group of Cur with
ester group of CA.
CT
cluding their structural (composition; hydrophilic or hydrophobic), could be the ideal material for wound treatment because of their ability
geometrical (shape, size and surface to volume ratio), morphological to take up the wound exudates besides providing a moist environment
properties (circular or ribbon-like flattened cross-section and existence for the healing process. The swelling ratio allows the pore size to be in-
of beads), pH, and temperature [16] [54–55]. The increased weight loss creased in diameter [57] ensuring that the cells not only attach to the
(%) in CA/PCL/Cur could be related to the high surface area of surface but could also penetrate inside the interpolymer network dur-
RE

nanofibers, resulting in a faster diffusion rate and a corresponding
build-up of protein degradation product to the medium. This can also
be attributed to the hydrophilicity of Cur which enhances the interac-
tion between CA/PCL nanofibers and the water molecules. Moreover,
Cur is likely to reduce its physical interactions between the polymer
chains causing a fast degradation rate. As noticed, the degradation rate
is increased by increasing the Cur content in the nanofibers which could
OR
ing MTT assay.
3.3. Drug release studies
The phenolic compounds of curcumin could be identified in the λmax
range ~300 to 550 nm in the UV–Vis absorption spectra, which arise
from the B-ring, band I, and band II [58]. Fig. 4d shows the curcumin

be related to hydrophilic nature as well as the thinner diameter of the
nanofibers.
3.2. Absorption capability by swelling behavior
An effective wound dressing must exhibit high absorbability, a cool-
ing sensation, a moisture environment, and other properties including
C
release profile determined from the UV–Vis spectra of both scaffolds up
to a period of 120 h. Four steps in the drug release profile was observed
for CA/PCL/Cur nanofiber scaffolds. At the initial hours (0–15 h), ini-
tial burst release has occurred. The nanofiber scaffolds loaded with cur-
cumin of 0.5% and 1.0% showed an initial burst release of 14% and
18%, respectively. Burst release is a phenomenon generally found in
drug delivery with diverse forms and concentrations [56]. The initial

biocompatibility, which is highly desired for the scaffold to act as a bar- burst release results from the immobilization of curcumin at the surface
rier from micro-organism and help to activate the platelets for the for- of the nanofiber scaffolds. In some cases, the initial burst release could
mation of the blood clot [56]. In this regard, the swelling behavior of also provide immediate pain relief, followed by prolonged release to
UN

the nanofibers scaffolds was investigated for up to 9 days to determine
their eligibility for wound healing application; the results are shown in
Fig. 4c. The swelling followed the order of CA/PCL/Cur
(1.0 wt%) > CA/PCL/Cur (0.5 wt%) > CA > CA/PCL > PCL. The
pure PCL nanofibers are non-absorbent and its water intake ability in
PBS showed the lowest swelling ratio percentage due to its hydropho-
bicity. It is clearly observed that the increase in curcumin content in
CA/PCL/Cur nanofibers leads to increased swelling, which could be as-
signed to the increased hydrophilicity with Cur content. The swelling
(%) for CA/PCL/Cur (0.5%) and CA/PCL/Cur (1.0%) was ~700 and
950%, respectively. Therefore, the curcumin-loaded nanofiber scaffolds
promote gradual healing [59]. After the initial burst release, the drug
release profile showed a gradual release reaching 45% and 60% of the
total drug of CA/PCL/Cur (0.5%) and CA/PCL/Cur (1.0%) respectively,
in the next 60 h. After this step, the release rate decreased during the
next 20 h with 60% and 70% of CA/PCL/Cur (0.5%) and CA/PCL/Cur
(1.0%), respectively, before reaching the equilibrium. The amount of
drug release reached an equilibrium of ~60–62% for the next 40 h in
the case of CA/PCL/Cur (0.5%) scaffolds, whereas the release rate was
gradually increased to 78% during the next 20 h in CA/PCL/Cur (1.0%)
scaffolds and stayed constant at 78–80% over the time of 120 h. At the
equilibrium, the amount of drug release from nanofibers was closely to

7
N.N. Suteris et al.
zero. After 120 h, there are 40% and 20% of curcumin remained in the
CA/PCL/Cur (0.5 wt%) and CA/PCL/Cur (1.0 wt%) nanofiber scaf-
folds, respectively. Higher the swelling ratio of the nanofibers leads to
better drug dissolution because of higher penetration of water mole-
cules inside the polymer. Therefore, the CA/PCL/Cur (1.0 wt%) having
the highest swelling ratio (950%), as observed in this study, showed the
higher curcumin release (78%) compared to that of CA/PCL/Cur
(0.5 wt%) with decreased swelling ratio (700%) and drug release
(60%). A first requirement for the development of a successful drug de-
livery is to demonstrate that the encapsulated drug is released suffi-
ciently and efficiently into the physiological environment [60]. The
highest cumulative drug release of 60% and 78% was observed for CA/
PCL/Cur (0.5%) and CA/PCL/Cur (1.0%) nanofibers, respectively.
3.4. Cell proliferation
The MTT assay was carried out to observe the biocompatibility and
cell proliferation effectiveness of the present nanofiber scaffolds [61].
Fig. 4e shows the cytocompatibility of 3T3 mouse fibroblasts on func-
tionalized biocomposite nanofibers scaffolds on the control, CA/PCL,
CA/PCL/Cur (0.5 wt%) and CA/PCL/Cur (1.0 wt%) on days 3, 6 and 9.
All nanofiber scaffolds showed a significant increase in the cell viability
from day 3 to day 9 due to the increase in the number of cells that pene-
trate and differentiate through the porous nanofibers. Day 3 until day 6,
the CA/PCL nanofiber scaffolds showed a significantly slow increase in
the rate of cell proliferation, which is probably because of the absence
of the cell recognition aspect in CA/PCL. This behavior made the CA/
PCL scaffolds to fail to form any focal adhesion between the nanofibers
and seeded fibroblast cells. On the other hand, CA/PCL/Cur (0.5 wt%)
ED
and CA/PCL/Cur (1.0 wt%) nanofiber scaffolds facilitated high prolif-
eration of fibroblast cells. The MTT assay test thereby shows that cur-
CT
cumin loaded scaffolds are non-toxic and biocompatible to initiate pro-
liferation of fibroblast cells for repairing the wound site. These incre-
ments are probably due to a higher cell density around the nanofibers
giving a better environment for fibroblast cells to differentiate. Overall,
such a condition is a potential approach due to its significantly pro-
longed development of the drug tolerance [62].
RE
The fibroblast cells are seeded on the biomaterial scaffolds, the cyto-
toxicity of the substances on the scaffolds may disturb the cell viability
and interaction with the scaffolds. The effect of CA, PCL and Cur on the
cell viability of the scaffolds can be analysed by using red CMTPX dye.
CMTPX dye is a cell-tracker, which is suitable for monitoring the live
cells in terms of the movement of the cells and morphology of the cells.
The dye enters the viable cell membrane, demonstrating a fluorescence
C OR
UN
Fig. 5. Expression of actin protein and morphology of fibroblast on day 6. (a) Expression of actin (i) control, (ii) CA/PCL, (iii) CA/PCL/Cur 0.5 wt%, and (iv) CA/
PCL/Cur 1.0 wt% (b) SEM micrographs for, (i) control, (ii) CA/PCL, (iii) CA/PCL/Cur 0.5 wt%, and (iv) CA/PCL/Cur 1.0 wt%.
8
International Journal of Biological Macromolecules xxx (xxxx) 1–10
that acts on cytosolic esterase enzymes. Differences in morphological
features were observed between CA/PCL scaffolds compared with CA/
PCL with curcumin as shown in Fig. 4f. The results were observed in flu-
orescent microscopy detecting live cells and observed more cells on the
CA/PCL/Cur scaffolds than CA/PCL scaffolds on day 6. This is due to
the hydrophobicity of the PCL, which reduces the attachment of the
cells. By adding cellulose as the major component and curcumin as the
F
drug, cells were well distributed around the scaffolds. CA/PCL/Cur
showed an increased number of cells with a better attachment and con-
OO
trolled arrangement along the nanofiber scaffolds compared to CA/PCL.
The results indicated that the CA/PCL/Cur nanofiber scaffolds are bio-
compatible and non-toxic to the cells in wound healing.
3.5. Actin cytoskeleton staining
Cytoskeletal filament is a protein of interlinking network that
shapes up the cells and is responsible for important functions of wound
PR
repair such as cell proliferation, attachment, and migration. Actin cy-
toskeleton networks subject wound contraction that help in tissue gran-
ulation and enable wound healing. Fibroblasts cells were stained with
Rhodamine phalloidin performed on the day 6 to reflect the physiologi-
cal phenotype of fibroblasts (Fig. 5a). It indicates the scattering of cyto-
plasmic actin inter-network bundles (red color) along with staining of
nucleus by DAPI (blue color). Distribution of actin was not clearly ob-
served in the CA/PCL scaffolds due to spatial scattering of actin, which
stress fibers with abnormal cell cytoskeleton morphology (Fig. 5a(ii)).
This may be due to nanofibers hydrophobic effect which can affect cell
cytoskeleton structure by not supporting for cell attachment and cell
spreading. In Fig. 5a(iii and iv), the CA/PCL/Cur 0.5 wt% and the CA/
PCL/Cur 1.0 wt% fibrous scaffolds show highly aligned scattering of
actin protein through the cell cytoskeleton with notable elongated fi-
broblast morphology and better cell to cell interactions compared to the
fibrous CA/PCL scaffold. The DAPI staining was used to detect any
changes in the nucleus of the fibroblast cells in vitro. Fibroblast cells on
CA/PCL/Cur scaffolds showed a highly aligned actin filament through
the cell cytoskeleton with a distinguished elongated fibroblast cell mor-
phology and a better cell performance compared to other scaffolds to
prove the wound healing.
Physical and chemical properties of fabricated biomaterial are re-
sponsible for the cell and scaffold interaction, cell communication, nu-
trients transport, molecular cell signaling and biological molecular ac-
tivity for cell proliferation and deposition of ECM; these aspects con-
tribute to the development of successful skin substitute for wound heal-
ing. A surface scanning (Fig. 5b) performed on the incubated scaffolds
N.N. Suteris et al.
with fibroblasts cells showed that the fabricated nanofibers scaffolds
support the cell morphology, attachment of cells, and cell spreading.
The SEM images in Fig. 5b are the fibroblast cells cultured on day 6. Fi-
broblast cells grown on the scaffolds (day 6) showed regular morphol-
ogy; however, CA/PCL/Cur with 1.0 wt% showed significantly in-
creased number of cells and irregular stretched morphology (Fig. 5b
(iv)). The CA/PCL/Cur 1.0 wt% shows improved spreading of the cells
and better cell attachment compared to other scaffolds. The CA/PCL/
Cur 1.0 wt% observed improved cell spreading, cell to cell communica-
tion and controlled extension of fibroblast along the nanofibers com-
pared to CA/CUR. The results in Fig. 5b (iii and iv) showed that CA/
PCL/Cur nanofiber as a wound dressing material can support high pro-
liferation of fibroblasts with controlled cell growth and spreading,
which are the native physiological characteristics of tissues. The scaf-
folds that contained Cur showed a better performance in terms of the
cell spreading and cell density. Generally, CA/PCL/Cur scaffolds are a
better candidate as wound dressing as these scaffolds show better cell
attachment and help the proliferation phase to take place with con-
trolled cell growth for the process of wound healing.
4. Conclusions
Cellulose acetate (CA) derived from an oil palm waste biomass
(empty fruit bunch) is used as the primary component (90 wt%) to fab-
ricate CA/polycaprolactone (PCL)/Curcumin (Cur) nanofiber scaffolds
for wound dressing applications. Scaffolds with two compositions of
Cur (0.5 and 1.0 wt%) are developed and compared with the character-
istics of CA, PCL, and CA/PCL. The curcumin contributed many proper-
ED
ties to the scaffold in addition to its role as a natural healing agent. The
CA/PCL/Cur scaffolds showed lower fiber diameter, increased hy-
drophilicity, higher swelling ratio, increased cell viability, and in-
CT
creased cell proliferation compared to the CA/PCL scaffolds. The results
proved that addition of curcumin improved the hydrophilicity, which
in turn originated from the hydrogen bonding between the polymers
and the curcumin. The cell proliferation showed that the drug-loaded
nanofibers are non-toxic to the cells for CA/PCL/Cur. The release of
curcumin from CA/PCL/Cur nanofiber scaffolds attribute to the fibrob-
RE
last cell enhancement, increased proliferation rate, and easy spreading
along the nanofiber scaffolds. The CA/PCL/Cur nanofiber scaffolds
proved potential material for drug carriers in skin tissue engineering
with excellent physical and biological properties for wound healing.
CRediT authorship contribution statement
OR
Nurul Nadirah Suteris: Conceptualization, Investigation, Visualiza-
tion, Writing – original draft, Writing - review & editing. Amina Yasin:
Investigation, Writing – original draft, Writing - review & editing. Izan
Izwan Misnon: Writing - review & editing. Rasidi Roslan: Writing - re-
view & editing. Farah Hanani Zulkifli: Writing & review. Mohd Hasbi
Ab Rahim: Writing & review. Jayarama Reddy Venugopal: Investiga-
tion, Visualization, Writing - review & editing. Rajan Jose: Conceptual-
C
ization, Investigation, Visualization, Writing - review & editing, Fund-
ing acquisition, Project administration, Supervision.
UN
Declaration of competing interest
The authors have no Conflict of Interest to declare.
Acknowledgments
The support of Ministry of Higher Education Malaysia (Ref. code:
FRGS/1/2019/STG01/UMP/01/1; UMP Ref: RDU 1901215 and FRGS/
1/2016/STG01/UMP/01/1; UMP Ref: RDU160112) is gratefully ac-
knowledged. Nurul Nadirah Suteris would like to acknowledge the Re-
search and Innovation department, Universiti Malaysia Pahang (UMP)
9
International Journal of Biological Macromolecules xxx (xxxx) 1–10
for the postgraduate research grant scheme (PGRS190355). Amina
Yasin would like to acknowledge the Post-Doctoral Fellowship from
UMP (http://ump.edu.my).
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://
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doi.org/10.1016/j.ijbiomac.2021.12.006.
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