720815

research-article2017
JOB0010.1177/0883911517720815Journal of Bioactive and Compatible PolymersJaganathan et al.

JOURNAL OF

Bioactive
and
Compatible
Polymers
Original Article

Journal of Bioactive and

Compatible Polymers
Production and 1–14
© The Author(s) 2017
hemocompatibility assessment Reprints and permissions:
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https://doi.org/10.1177/0883911517720815
DOI: 10.1177/0883911517720815
of novel electrospun journals.sagepub.com/home/jbc

polyurethane nanofibers loaded

with dietary virgin coconut oil
for vascular graft applications

Saravana Kumar Jaganathan1,2,3,

Mohan Prasath M4, Ahmad Fauzi Ismail5,
Manikandan A6 and Gomathi N7

Abstract
To develop biodegradable polymer scaffolds suitable for vascular tissue engineering applications,
the bioengineering community has invested an extensive effort. The most common cause for the
failure of vascular graft scaffolds is thrombosis. In this work, the scaffold based on polyurethane
and virgin coconut oil was produced by electrospinning process for vascular tissue engineering
applications with improved antithrombogenicity. The diameter of this electrospun polyurethane/
virgin coconut oil composite was found to be reduced in the range of 886 ± 207 nm compared
to pristine polyurethane which was in the range of 969 ± 217 nm. The Fourier transform infrared
spectroscopy analysis revealed the interaction between polyurethane and virgin coconut oil as

1Department for Management of Science and Technology Development, Ton Duc Thang University, Ho Chi Minh City,
Vietnam
2Faculty of Applied Sciences, Ton Duc Thang University, Ho Chi Minh City, Vietnam
3IJN-UTM Cardiovascular Engineering Center, Department of Clinical Sciences, Faculty of Biosciences and Medical

Engineering, Universiti Teknologi Malaysia, Skudai 81300, Johor, Malaysia

4Faculty of Biosciences and Medical Engineering, Universiti Teknologi Malaysia, Skudai 81300, Johor, Malaysia
5Advanced Membrane Technology Research Centre (AMTEC), Universiti Teknologi Malaysia, Skudai 81310, Johor,

Malaysia
6Department of Chemistry, Bharath Institute of Higher Education and Research, Bharath University Chennai 600073,

Tamil Nadu, India

7Department of Chemistry, Indian Institute of Space Science and Technology, Thiruvananthapuram 695547, India

Corresponding author:
Saravana Kumar Jaganathan, Department for Management of Science and Technology Development, Ton Duc Thang
University, Ho Chi Minh City, Vietnam.
Email: saravana@tdt.edu.vn
2 Journal of Bioactive and Compatible Polymers 00(0)

indicated by phase shifting of CH bond along with the formation of hydrogen bond. The contact
angle measurement of fabricated composites was found to be increased owing to hydrophobic
nature and also exhibited enhanced thermal stability as noted in thermogravimetric analysis. The
atomic force microscopy analysis insinuated the increased surface roughness of the composite in
comparison with the pure polyurethane. Developed scaffold resulted in delayed blood clotting
as revealed by activated partial thromboplastin time and partial thromboplastin time assay. The
hemolytic index of fabricated composites was found to be low indicating the enhanced safety of
red blood cells. Hence, the newly developed nanofibrous composite scaffold could open the door
for a suitable alternative for vascular graft applications.

Keywords
Polyurethane, virgin coconut oil, nanocomposite, vascular graft applications, electrospun

Introduction
In cardiovascular application, for repairing the damaged blood vessel, the use of prosthetic vascular
graft material such as dacron and polytetrafluoroethylene (PTFE) is required.1–3 In replacement of
aorta or iliac vessels, the prosthetic vascular graft materials were fairly comparable to the native
blood vessel, but for smaller-diameter applications like bypasses of the lower leg or coronary arter-
ies, these materials do not perform well.4–6 The reason might be trouble associated with thrombosis,
graft occlusion, and infection.7–9 Among these issues, the thrombosis was vital for vascular graft
failure. Hence, the fabricated membrane should have essential characteristics like thromboresistant
and anticoagulant nature in order to prevent the graft failure.10 The fabricated scaffold comes in
direct contact with red blood cells (RBCs) and it should not induce any damage to the RBCs.
Thromboresistant properties are assessed by the measurement of activated partial thromboplastin
time (APTT) and prothrombin time (PT). These two time points are the good indicators of intrinsic
and extrinsic pathways, respectively. The hemolytic percentage may serve as a yardstick for estimat-
ing the damage to RBCs.11 Many articles have predicted poor tissue response and integration of
PTFE after implantation. Many types of researches utilized tissue-engineered vascular grafts in
fabricating small-diameter (<5 mm) vascular grafts to overcome these complications.9,12,13 Recently,
tissue-engineered vessels have become a promising solution to overcome the limitations associated
with current therapies. This technique utilizes the scaffold incorporated with vascular cells to build
functional blood vessels. The incorporation of endothelial cells onto scaffold has maintained the
patency, facilitated graft maturation, and promoted remodeling when implanted in vivo.14–16
The tissue-engineered scaffolds prepared by decellularizing animal vessels possessed some dis-
advantages like limited length of unbranched segments, inconsistencies in composition and the
potential risk of xenogenic pathogens that limits the clinical setting.17 The growth of electrospin-
ning technology has favored the fabrication of nanofiber-based scaffolds that facilitate cell growth
and also develop scaffold materials with distinct composition, which allows for controlled degra-
dation during remodeling.18–20 Electrospinning has been known since the 1930s, but the utilization
of electrospun fibers with the high surface area in applications like high-performance filters and for
scaffolds in tissue engineering was recently emerged.21–23 Electrospinning is the technique that
involves applying the high voltage to polymer melts which are drawn into nanofibers at the collec-
tor end. The fiber diameter was easily controlled in electrospinning technique by regulating the
voltage, polymer solution concentration, and solvent composition. In addition, electrospinning is
easy and it is possible to obtain typical fiber diameters in the range from 100 to 1000 nm. Electrospun
nanofibers have small pore size and high surface area suitable for biomedical applications.24,25 In
Jaganathan et al. 3

this research, tecoflex EG-80A which is a poly-ether-based medical-grade polyurethane (PU) was
used to fabricate the nanofibers. Owing to good barrier properties and oxygen permeability, the PU
is frequently used in making scaffold for tissue engineering applications.
Virgin coconut oil (VCO) is the finest and purest grade of VCO available. VCO is edible which
is extracted from the kernel or meat of mature coconut (Cocos nucifera). It will oxidize slowly and
is resistant to rancidification lasting up to 6 months at 24°C without spoiling owing to high saturated
fat content.26 It was widely used in many food and cosmetic preparations because of its medicinal
benefits. The other applications of VCO were feedstock for biodiesel, herbicides, and lubricants.27–29
The fabricated composite comprises VCO, which is a natural ingredient and a food; hence, the toxic-
ity rendered by adding a synthetic agent is greatly reduced. In a recent study, Yeap et al. investigated
the antistress and antioxidant nature of VCO. They found that VCO-administered mice was able to
restore the oxidative stress effectively by exhibiting higher levels of brain antioxidants and lower
levels of brain 5-hydroxytryptamine. Moreover, the levels of serum cholesterol, triglyceride, glu-
cose, and corticosterone were also lowered indicating the antistress nature of VCO.30 This invigor-
ated the interest of using VCO oil in this study. Moreover, in a recent study, it was concluded that
VCO has no detrimental effect on blood pressure and inflammatory markers.31 Further to our moti-
vation, Amna et al. reported that the virgin olive oil–blended PU scaffold showed significant cell
viability resulting in enhanced cell adhesion and behavior. They attributed this behavior to the anti-
oxidant nature of the olive oil.32 Hence, in this study, PU is blended with VCO for the first time to
examine the putative role in influencing the blood compatibility. The aim of this research is to
develop a novel nanocomposite based on PU and VCO using a single-step electrospinning process
and to investigate the physicochemical characterization of the developed nanocomposite.
Furthermore, blood compatibility of the composite along with the pure PU nanofibers will be
assessed to understand the compatibility of the scaffold with the surrounding tissues.

Materials and methods

In this work, the polymer used was tecoflex EG-80A a medical-grade thermoplastic PU purchased
from Lubrizol, USA. N,N-dimethylformamide (DMF) was obtained from Merck Millipore,
Germany. The commercially available VCO was obtained locally. The chemicals phosphate-buff-
ered saline (PBS, Biotech Grade) and sodium chloride physiological saline (0.9% w/v) were sup-
plied by Sigma-Aldrich, Malaysia. The reagents utilized in this work for APTT and partial
thromboplastin time (PT) assay were rabbit brain activated cephaloplastin, calcium chloride
(0.025 M), and thromboplastin (Factor III) obtained from Diagnostic Enterprises, India.

Preparation of nanocomposite
To prepare PU melt mix, initially, 480 mg of PU beads were dissolved in 6 mL of DMF followed
by magnetic stirring for 24 h at room temperature to obtain a homogeneous solution of concentra-
tion 8% (w/v). Similarly, for VCO mix, 400 µL of VCO was mixed with 4.6 mL of DMF to make
8% v/v solution and stirred for 1 h minimum to obtain a homogeneous solution. Finally, the PU/
VCO nanocomposite was obtained by mixing VCO solutions in PU at a ratio of 8:2 under rigorous
stirring for 60 min.

Fabrication of PU and nanocomposite dressing

The nanocomposite of pure PU and PU/VCO fibers was fabricated using electrospinning tech-
nique. To prepare the composites, initially, the prepared solutions of PU and composite was injected
4 Journal of Bioactive and Compatible Polymers 00(0)

inside the plastic syringe of 10 mL with an 18-G stainless steel needle and placed in the syringe
pump (SP20, NFiber). Then, NFiber high voltage unit was utilized to supply the voltage to the
syringe for obtaining nanofibers. The electrospun nanofibers were fabricated and collected on a
static drum collector covered with aluminum foil. After several efforts, the PU was successfully
obtained in the form of nanofibers at a flow rate of 1.0 mL/h with an applied voltage of 10 kV. Since
the incorporation of VCO will reduce the viscosity of the nanocomposite solution, the flow rate
and voltage were adjusted to 0.50 mL/h and 7 kV, respectively, to obtain a steady jet of flow. The
distance of collector was 16 cm which was constantly maintained for both samples. The residual
DMF solvent present after electrospinning was removed by drying the developed scaffold under
vacuum at room temperature for 24 h.

Physicochemical characterization
Scanning electron microscopy micrographs
The fiber distribution and surface morphology of the electrospun PU and the VCO nanocomposite
fibers were analyzed using a Hitachi Tabletop SEM unit (TM3000). The fabricated samples were
in need of gold coating before obtaining the photomicrographs. The diameter size distribution in
the developed membranes was calculated using ImageJ (National Institutes of Health, Bethesda,
MD) software by measuring at least 30 individual fibers randomly. The mean and standard devia-
tion of the fiber diameter distribution was obtained from ImageJ software.

Attenuated total reflectance Fourier transform infrared spectroscopy analysis

The absorption bands of the electrospun PU and the VCO nanocomposite were analyzed using the
attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy unit. To identify
the peaks in the spectra of PU and VCO nanocomposite, a small piece of the sample was cut and
placed on the sensor surface and then the spectra were examined. In the interim, the IR spectra of
VCO were also obtained by placing a drop of the sensor surface. To investigate characteristic
peaks, each sample spectra were recorded over the range of 600–4000 cm−1 at 32 scans per minute
and averaged at the resolution of 4 cm−1. The ATR crystal used was Zinc/Selenium which was cou-
pled with the NICOLET IS5 spectrometer. After recording, the spectra were baseline corrected and
integrated using Spekwin 32 software to identify the absorption band peaks and differences.

Contact angle measurement

The measurement of contact angle for electrospun PU and the VCO nanocomposite was calculated
using the VCA Optima contact angle measurement unit. For contact angle measurements, the sam-
ples were cut in the size of 1 × 5 cm2 and placed on the equipment surface for measuring the contact
angle. After placing the sample, the syringe was loaded with water and then a droplet with a size of
2 µL was formed to fall on the test membrane. Within few seconds, the static image of the liquid
deposition within the membrane was obtained with the aid of high-resolution video camera. The
contact angle measurements of samples were repeated for many different trails and the manual
contact angle was measured through computer-integrated software.

Thermogravimetric analysis
The thermal degradation of electrospun PU and the VCO nanocomposite was investigated using
the PerkinElmer TGA 4000 unit. The samples weighing 3 mg were placed in an aluminum pan and
Jaganathan et al. 5

the experiment was performed under a dry nitrogen atmosphere in the temperature range of
30°C–1500°C at an ascending rate of 10°C/min. Then, the residual weight of the sample was meas-
ured at each temperature point and the measured experimental values were transferred to an excel
sheet. Then, the thermogravimetric analysis (TGA) curve and the equivalent derivative weight loss
curve (DTG) were drawn using OriginPro 8.5 software.

Atomic force microscopy

To analyze the surface roughness of sample, atomic force microscopy (AFM) was used.
Furthermore, to attain a 3D image of the sample surface, JPKSPM data processing software was
utilized. To perform AFM analysis, the sample with a size of 1 × 1 cm2 was cut from the developed
membrane and placed on the AFM equipment (Nanowizard, JPK instruments). The scanning was
performed at room temperature in the normal atmosphere to measure the surface roughness (Ra).
The images were acquired with the scanning size of 20 µm × 20 µm and were recorded in the
medium mode with 256 × 256 pixels.

Hemocompatibility assessment of the scaffold material

Ethical statement and collection of blood samples. All the experimental procedures implicated in the
handling of blood were approved by Faculty of Biosciences and Medical Engineering, Universiti
Teknologi Malaysia with ref no: UTM.J.45.01/25.10/3Jld.2(3). The blood was collected from
healthy adults who were educated about the risk and benefits of the blood donation. The blood was
collected via venipuncture after getting a signature in the consent form. The collected blood was
anticoagulated with acid–citrate–dextrose (ACD) (56 mM sodium citrate, 65 mM citric acid,
104 mM dextrose) at a ratio of 9:1 (blood/citrate). Finally, citrated blood was centrifuged at 3000 r/
min for 15 min to extract platelet-poor plasma (PPP).

APTT assay. Initially, the fabricated PU and PU/VCO scaffold were cut into square samples of
dimension 0.5 × 0.5 cm2. The assay was carried out in triplicate, so three square samples of each
type were introduced into 96-well plates and gently washed with deionized water. The samples
were incubated in PBS at 37°C for 30 min before starting the assay. To begin with, 50 µL of the
obtained PPP was placed on the sample and incubated for 1 min at 37°C. After, 50 µL of rabbit
brain cephaloplastin reagent was incorporated and incubated for 3 min at 37°C. Finally, the reac-
tion mixture was activated by adding 50 µL of CaCl2 and gently stirred with a sterile steel needle.
The time taken for the formation of the white fibrous clot was measured using a chronometer.

PT assay. For PT assay, the fabricated nanofibrous membrane was cut into square samples as
described in the “APTT assay” section, and this test was also performed in triplicate. The samples
were washed with deionized water and incubated in PBS for 30 min at 37°C. It was further incu-
bated in 50 µL of PPP at 37°C for 1 min and then 50 µL of NaCl–thromboplastin reagent (Factor III)
was added and gently stirred with a sterile steel needle until clot formation.

Hemolysis assay. The hemolysis assay was performed using citrated whole blood to determine the
effect of fabricated membranes on RBCs. Initially, both PU and bio-nanofibrous samples (1 × 1 cm2)
were soaked in physiological saline (0.9% w/v) at 37°C for 30 min. Then, the soaked samples were
incubated with a mixture of aliquots of citrated blood and diluted saline at a ratio of 4:5 (v/v%) for
1 h at 37°C. For complete hemolysis, the whole blood was diluted with distilled water (4:5) along
with physiological saline solution to make positive and negative controls, respectively. After incu-
bation, the samples were taken and the mixtures were centrifuged at 3000 r/min for 15 min. Using
6 Journal of Bioactive and Compatible Polymers 00(0)

pipette, the clear supernatant was carefully taken out and the absorbance of each sample was meas-
ured at 542 nm to record the amount of hemoglobin released, which directly represents RBC dam-
age. Finally, the percentage of hemolysis or hemolytic index was calculated using the formula33

Hemolysis ratio ( HR ) = ( TS − NC ) / ( PC − NC ) × 100

where TS, NC, and PC are measured absorbance values of the test sample, negative control, and
positive control at 542 nm, respectively.

Statistical analysis
All experiments were conducted thrice independently. Unpaired t-test was used to determine sta-
tistical significance. The results obtained from all experiments are expressed as mean ± SD. In the
case of qualitative experiments, a representative of three images is shown.

Result and discussion

The scanning electron microscopy (SEM) image of prepared PU/VCO composites and pure PU are
shown in Figure 1(a) and (b). The fabricated nanocomposites showed randomly oriented and finer
dispersion of nanofibers in the matrix as noted in SEM image. ImageJ analysis of prepared samples
showed the mean fiber diameter and standard deviation of 886 ± 297 nm and for pure PU it was
969 ± 217 nm respectively. The distribution curve of fiber diameters for pure PU and PU/VCO is
shown in Figure 2(a) and (b). PU/VCO composites had decreased fiber diameter compared to pure
PU fibers. The reason for reduced fiber diameter was due to an increase in conductivity of solution
mix owing to the incorporation of VCO in the polymer matrix. Balaji et al.33 reported an increase
in the conductivity resulting in the decreased diameters of fabricated nanocomposites. Kumbhar

Figure 1. SEM images of (a) polyurethane and (b) polyurethane/VCO composites.

Jaganathan et al. 7

Figure 2. Fiber distribution curve of (a) polyurethane and (b) PU/VCO composites.

et al. observed the highest proliferation of fibroblast cells at the range of fiber diameter 600–
1200 nm compared with the other diametric ranges. The author also reported that the composites
promoted collagen III expressions in the same fiber diameter range.34 The fiber diameter results of
our prepared composite scaffolds were found to be within this range, and it may be suitable for
wound remodeling resulting in the highest proliferation of fibroblast cells. Dan et al. developed
polycaprolactone (PCL)/gelatin scaffold for cardiac tissue engineering incorporated with different
weight concentrations (0%–30%) of polypyrrole (PP). The fiber diameter of fabricated nanocom-
posites was found to be reduced and showed enhanced cell attachment, proliferation, and expres-
sion of cardiac-specific proteins compared to control.35 In our investigation, the fiber diameter was
observed to be following similarly decreased pattern and which may allow more proliferation of
cardiac cells for new tissue growth.
The IR spectrum indicated the functional groups of PU/VCO and prepared PU/VCO nanocom-
posites are denoted in Figure 3. The spectrum indicated peak curve at 3323 cm−1 indicating the
presence of NH stretching of an aliphatic primary amine and the peaks at 1597 and 1531 cm−1 due
to vibration of NH. The characteristic bands at 2939 and 2854 cm−1 were due to CH stretching, and
the band at 1413 cm−1 indicates the vibrations of CH. For C=O stretching of carboxylic groups, the
twin peak was identified at spectra 1730 and 1703 cm−1, respectively. The peaks of C–O stretching
corresponding to alcohol groups was found at 1221, 1104, and 770 cm−1.36,37 In VCO IR spectrum,
the characteristic sharp peak bands at 2922 and 2853 cm−1 were due to CH stretch and the peak
corresponding to C=O stretching was observed in the band of 1743 cm−1. The peaks at 1470 and
1151 cm−1 indicated the vibrations of CH and C–O stretch groups, respectively. There were no
additional peaks present in PU/VCO, but there was an apparent decrease in the intensity revealing
the interaction between PU and VCO components due to the formation of hydrogen bonds.38 The
interaction between PU and VCO is also identified by slight shifting of CH stretching band at
2939 cm−1 in PU to 2928 cm−1 in PU/VCO mats.39
The wettability of PU and prepared PU/VCO composites were obtained through contact angle
measurements. The prepared PU/VCO composites depicted a mean contact angle of 118° ± 1.55°,
whereas, for pure PU, the water contact angle was found to be 86° ± 1.91°. It was clearly proven
that the PU/VCO nanocomposite samples will behave like hydrophobic as the water contact angle
exceeded 90°. The smaller fiber diameter resulted in increased water contact angle as concluded in
some research articles reported by Cui et al.40 and Ceylan.41 Hence, we assume that our reduced
fiber diameter in the PU/VCO composite may be the reason for the increasing contact angle
observed.
8 Journal of Bioactive and Compatible Polymers 00(0)

Figure 3. FTIR analysis of polyurethane, VCO, and PU/VCO composites.

Thermal stability of VCO loaded on PU composites was evaluated by TGA. From the results
obtained, it was found that the pure PU starts to degrade at 273.24°C, whereas the sample of PU/
VCO composites degrade at 289°C as shown in Figure 4(a) and (b). The higher thermal stability
behavior of PU/VCO composites may be due to incorporated constituents of VCO into the polymer
matrix. The residual weight loss of pure PU and the composite material calculated at 900°C was
found to be 1.91% and 6.69%, respectively, as denoted in Figure 5(a) and (b). The results showed
that the addition of VCO into PU has improved the thermal stability of prepared composites. Kuan
et al.42 fabricated and investigated the thermal properties of multi-walled carbon nanotubes
(MWCTs)/PU composites and reported that the addition of carbon nanotubes into the polymer
matrix had improved the thermal stability of the PU and our experimental results were similar to
their findings.
The surface roughness of fabricated composite and pure PU patch was determined using AFM
analysis as shown in Figure 6(a) and (b). From the results obtained, the surface roughness of the
pristine PU was found to be 723 nm, and for the prepared composite patch, it was found to be
986 nm (Ra), respectively. The increased surface roughness was due to incorporated constituents of
VCO in the polymer matrix. Nikpour et al.43 synthesized and characterized nanocomposite based
on hydroxyapatite and chitosan materials for medical engineering applications and found that the
addition of hydroxyapatite into chitosan membrane resulted in an increase in the surface roughness
and our fabricated composites displayed similar to those observations.
The blood compatibility of pure PU and PU/VCO was studied using APTT and PT which
denotes the intrinsic and extrinsic pathways associated with blood clotting, respectively. From the
APTT results, the blood clotting time of PU/VCO composites was found to be higher than the
pristine PU. The prepared PU/VCO nanocomposites showed blood clotting time with a mean value
of 220 ± 6.2 s, while for PU membrane, the blood clotting was found to be 157.3 ± 5.55 s as noted in
Figure 7. It was found that the fabricated nanocomposites showed delayed clotting time indicating
that nanocomposite surface was better than the PU and delays the activation of intrinsic pathway
Jaganathan et al. 9

Figure 4. TGA analysis of (a) polyurethane and (b) PU/VCO composites.

Figure 5. Weight residue percentage of (a) polyurethane and (b) PU/VCO composites.

resulting in the enhancement of anticoagulant nature of the PU composite. Similarly, in PT assay,

the prepared PU/VCO composites insinuated delayed clotting time of about 53.67 ± 2.5 s compared
to pure PU where the blood clotting time was found to be 38.33 ± 1.15 s as shown in Figure 7. The
delayed clotting time may be due to the enhanced surface roughness as observed in our studies.
Milleret et al. found that the smaller fiber diameter will be conducive for blood compatibility. In
their work, they used two polymers, namely, degarapol and poly (lactic-co-glycolic acid) (PLGA)
for fabricating scaffolds with different diameters and observed that the smaller fiber diameter
showed the delay in blood clotting which seems to be in concurrence with our findings.44 Moreover,
the VCO constituents in PU matrix might also play an important role in delaying the clotting time.
Next, the hemolytic percentage was calculated for fabricated PU/VCO composite to investigate the
safety of the fabricated patch against RBCs. The hemolytic percentage was obtained by recording
the absorbance of obtained supernatant after blood reacts with the composite patch at 542 nm.
10 Journal of Bioactive and Compatible Polymers 00(0)

Figure 6. AFM analysis of (a) polyurethane and (b) PU/VCO composites.

Figure 7. APTT and PT assay of polyurethane and PU/VCO composites.

From the hemolytic assay, compared to PU/VCO composites, the absorbance value of PU was
found higher indicating lysis of erythrocytes by pristine PU. The prepared PU/VCO composite
shows hemolytic index of 0.63%, while for pure PU membrane, the index was found to be 2.73%,
respectively. According to ASTMF756-00(2000) standard, when the hemolysis percentage was
above 2%, it was considered as hemolytic material and the percentage below 2% was considered
as non-hemolytic material.45 The hemolytic percentage results of fabricated patches were found to
be 0.63% which was below 1% and hence it will behave like non-hemolytic material. Chen et al.
investigated the blood compatibility of carbon/TiO2 nanocomposite and reported that the
Jaganathan et al. 11

nanocomposite exhibited improved anticoagulant function and better blood compatibility. They
concluded that the increase in anticoagulant function and improved blood compatibility was due to
the hydrophobic nature of the TiO2 nanowire arrays.46 Our developed nanocomposite was found to
be hydrophobic which favors enhanced blood compatibility and may be suitable for cardiac engi-
neering scaffolds.
In order to serve as a scaffold in cardiovascular applications, the first and foremost criteria is
that scaffold must possess thromboresistant and anticoagulant nature in order to prevent the vascu-
lar graft failure. The measure of blood compatibility would determine the thromboresistant and
anticoagulant nature of the developed scaffold. Developed nanocomposite with the addition of
VCO into the PU resulted in 40% enhancement in APTT time compared to pristine PU. APTT is a
versatile indicator how the fabricated nanocomposite interferes with the intrinsic pathway. Since
there is a significant delay in APTT, it indicates the anticoagulant nature of the nanocomposite.
Furthermore, when blending the VCO into the pure PU, the hemolytic index was observed to be
low indicating it as a safe material compared to pure PU. There was 77% reduction in the hemo-
lytic index value of the prepared nanocomposite. All these indicate that it is a good alternative
scaffold which might be suitable for cardiovascular graft applications in reducing the graft failure
owing to enhanced blood compatibility and non-hemolytic behavior.
Furthermore, the scope of usage of this scaffold was discussed. The scaffold can be surface
modified to add further functionalities. Plasma surface modification results in enhanced endothe-
lial cell adhesion47 and improved antimicrobial efficiency.48 Hence, plasma-modified PU-VCO
scaffold may provide a suitable environment for endothelial cell adhesion resulting in improved
properties facilitating the vascular tissue growth. Recently, Wu et al.49 reported that the ideal car-
diovascular patch material should possess long durability, resistance to degradation and infection,
and non-toxicity. The increased thermal stability of our developed patch indicates the integrity and
durability signifying our developed nanocomposites might be a suitable candidate for cardiovascu-
lar patches. Furthermore, the VCO showed better antimicrobial and non-toxic behaviors,50 which
might also have great potential toward the cardiovascular tissue engineering. Moreover, developed
PU-VCO can be considered as scaffolds for other tissue engineering applications like bone, skin,
and dental due to its promising physicochemical and blood-compatible properties. However, their
cytocompatibility and antimicrobial effects are need to be studied to realize its potential. This is
vital for understanding how the included components may react with target cells at the particular
location where the fabricated nanocomposite is intended to use.

Conclusion
From overall results, the PU composite patch prepared using electrospinning technique displayed
reduced diameter compared to the pure PU. The presence of VCO in the polymer matrix has
decreased the intensity of the PU/VCO composites as revealed by FTIR. The contact angle of fab-
ricated nanocomposites was found to be increased indicating hydrophobic behavior and possess
enhanced thermal stability as evident by TGA. The existence of VCO in the polymer matrix has
shown the increase in the surface roughness value compared to control confirmed by AFM analy-
sis. The APTT and PT assay revealed that the fabricated nanocomposites showed delayed blood
clotting indicating the antithrombogenicity nature of the composite patch in comparison with the
pure PU patch. Moreover, the hemolytic percentage of fabricated PU/VCO composites was found
to be low compared to control signifying the protective effect of the composite against RBCs.
Hence, we suggest that the newly developed nanocomposites based on PU incorporated with VCO
having outstanding properties like better surface and blood compatibility could be exploited as a
substitute scaffold for the vascular grafts.
12
Declaration of conflicting interests
The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publi-
cation of this article.
Funding
This work was supported by the Ministry of Higher Education Malaysia with the grant no. Q.J130000.
2545.14H59.
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