Original article

Textile Research Journal

2018, Vol. 88(24) 2800–2809

Studies of mechanical properties of ! The Author(s) 2017

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electrospun zein/poly(e-caprolactone) DOI: 10.1177/0040517517732083
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composites and antibacterial properties
against Listeria monocytogenes strains
of zein/poly(e-caprolactone)/
poly(e-lysine) films

Xiyu Hu1, Liang liang Yang2, Kaijun Wang2, Yanli Zixuan Wei2,
Hongwu Deng2 and Jiang Dong1

Abstract
Ultra-fine fibers of zein/poly(e-caprolactone) (PCL) composites were prepared by oriented electrospun technology to
enhance the toughness and ductility of zein. The mechanical, morphological and thermal properties of fibrous films of
zein/PCL were studied by texture analysis and nanoindentation tests, scanning electron microscopy and dynamic mechanical
analysis. Wettability of fibrous films was also measured and discussed. The mechanical properties of these fibrous films were
significantly improved with the addition of PCL. Research on the antibacterial activities of poly(e-lysine) (EPL) loaded films of
zein/PCL/poly(e-lysine) shows that the films could inhibit the growth of Listeria monocytogenes. Films of zein/PCL blend
systems with a hydrophilic surface do not dissolve in water and have excellent toughness and ductility. This is very favorable
for applications in biodegradable materials, food packing materials, facial masks and as antimicrobial films for wound dressing.

Keywords
hydrophilicity, poly(e-caprolactone), poly(e-lysine), zein

In recent years, research on agriculture-based materials
has been viewed with tremendous interest due to envir-
onmental issues and agronomics. Agriculture-based
materials also serve as alternatives for petroleum-
based synthetic polymers. As energy demands world-
wide soar and petroleum resources shrink, the energy
sector is focusing more on the conversion of biomass
into useful products. However, the advantage of green
biomass can be realized only if the ‘waste’ products are
efficiently utilized. A number of biopolymers, including
soybean proteins, casein, cellulose, poly(lactic acid),
chitosan, poly(e-caprolactone), zein and starch have
been investigated as structural materials.1–4 They have
been found to be potentially useful for packaging appli-
cations, facial mask, sustained-release drug carriers and
cellular growth. However, biopolymers are brittle and
have poor ductility. Extreme brittleness interferes with
the processing of biopolymeric materials and renders bio-
polymeric films less suitable for wrapping applications.
Zein consists of alcohol-soluble proteins (prolamines)
and is extracted from the maize kernel.5 It has been stu-
died as a biopolymeric carrier for gradual release of
encapsulated compounds.6–7 It is insoluble in water but
soluble in 5590% (v/v) aqueous ethanol.8 It shows
great potential in a number of industrial applications,
including food packaging. Although zein has a long his-
tory of use as a coating material for candies, medical
tablets and paper,9 other applications are still limited,
1
Engineering Research Center of High Performance Plastics, Ministry of
Education, College of Chemistry, Jilin University, Changchun 130012, P.R.
China
2
College of Plant Science, Jilin University, Changchun 130062, P.R. China
Corresponding author:
Jiang Dong, Jilin University, Qianjin Street 2699, Chang Chun, Jilin 130012,
China.
Email: jiangdonghxy@163.com
Hu et al.
mainly due to the undesirable properties of zein-based
materials.10–12 Like many other proteins, zein, in the
absence of plasticizers, forms brittle films with poor flexi-
bility that cannot withstand industrial processing.
In order to improve the mechanical properties of zein,
plasticizers13–14 are always added to modify its supra-
molecular structure. This decreases the intermolecular
interactions between zein protein chains, and effectively
improves flexibility, extensibility and toughness of zein.
Poly(e-caprolactone) (PCL) is a semi-crystalline ali-
phatic polyester having a melting temperature (Tm) of
approx. 60 C, and glass transition temperature (Tg) of
approximately 60 C. It is one of the most important
biodegradable polymers that has gained interest due to
its technological properties and inherent biocompatibil-
ity. Therefore, it has been widely applied in tissue
engineering,15–17 as drug carriers, engineered skin18
and scaffolds for the support and growth of fibroblasts
and osteoblasts,19–20 among other applications.
However, its hydrophobicity often leads to unfavorable
cell adhesion and growth.21 Hence, in this article, a
series of ultra-fine fibers of zein/PCL systems films
were prepared by oriented electrospun technology,
with the aim of improving the mechanical properties
of zein and also enhancing the superficial hydrophilicity
of PCL. Mixing two or more polymers to produce
blends or alloys is a well-established strategy for achiev-
ing specific physicochemical properties. It is a simple
and economical method, without the need to synthesize
a new functional polymer.22 At the same time, the cyto-
compatibility of PCL should be further improved.
Poly(e-lysine) exhibits antimicrobial activities against
a wide spectrum of microorganisms, including Gram-
positive and Gram-negative bacteria, yeasts and
molds.23 Listeria monocytogenes is a Gram-positive
pathogenic bacterium, responsible for serious infections
in the elderly, pregnant women, infants and immuno-
compromised individuals, and is a deadly foodborne
pathogen of vital importance to public health and the
food industry.24 Hence, a poly(e-lysine) (EPL) loaded
film of zein/PCL (70/30) was prepared and tested for its
antibacterial activity against L. monocytogenes. EPL-
loaded films of zein/PCL were able to inhibit the
growth of L. monocytogenes. Our main aim was to
develop a biocompatible and biodegradable material
so as to provide more options for materials for food
packaging, facial masks, wound dressing, drug delivery,
controlled release and other biological functions.
Experiment
Materials
Zein (purity 98%), PCL (Mn ¼ 70,000) and anhydrous
organic solvents, such as N,N-dimethylformamide,
2801
acetic acid and dichloromethane were obtained from
Sigma-Aldrich (Milwaukee, WI, USA). Poly(e-lysine)
(Mn ¼ 3000) was purchased from Zhejiang Silver-
Elephant Bioengineering Co. Ltd. (Zhejiang, China).
The solvents and materials were used without further
purification, unless specified. L. monocytogenes were
obtained from the Veterinary Science Department at
Jilin University, China. The trypticase, pepton, yeast
extract, dipotassium phosphate, glucose and nutrient
agar were purchased from Beijing AoboXing
Biological Technology Co., Ltd. (Beijing, China).
Methods and instruments
Electrospinning equipment. Self-regulated electrospinning
equipment was used for the preparation of randomly
oriented and symmetrical fibers of zein/PCL blend
systems. Scheme 1 shows the design of the oriented
electrospinning set-up, consisting of a syringe pump
(New Era Pump Systems Inc., New York, USA), a
high voltage supplier (030 kV) and a grounded collec-
tor. Two fiber collectors, aluminum foil and drum cov-
ered with a self-regulating copper wire were used to
collect the oriented fibers.
Morphological analysis. Morphologies of pure zein, pure
PCL and ultra-fine fibers of zein/PCL and zein/PCL/
EPL were directly observed using SEM (Hitachi,
SU8020, Ice emission, Bruker Spectrometer, Tokyo,
Japan) that operated at an accelerating voltage of
15 kV. The films were sputter-coated with gold for
2 min. The diameters of the electrospun fibers were
determined using Nano Measurer Software (Nano
Measurer 1.2, Fudan University, Shanghai, China)
and the average fiber diameters were calculated.
Scheme 1. Oriented electrospinning equipment.
2802
Wettability measurement. Static water contact angle
(WCA) measurements were carried out using VCA
Optima Contact Angle Analysis system (AST products,
Inc., Boston, USA; power: 110220 V Optima
1020021024 ) at room temperature. The images were
obtained using a CCD camera immediately after the
water drops were deposited onto the film surfaces of
zein/PCL/EPL with different compositions. An average
of at least five measurements for each sample, made at
five different regions of the film, was recorded.
Dynamic mechanical analysis (DMA). The analyses were
carried out on a TA Instruments DMA Q800 (New
Castle, Delaware, USA) at a heating rate of 5 C/min,
with a load frequency of 1 Hz in air, and Tg was deter-
mined from the peak temperature of tan .
Mechanical properties. Tensile tests were carried out on a
Texture Analyzer (CT3 Texture Analyzer, Ametek
Brookfield, USA). The tensile strength, modulus and
elongation at break were reported as average values
for at least 20 samples; the data for the stress–strain
curves shown in Figure 1 are close to the average
values. The tensile specimens were cut using a pump
knife (the effective dimensions were
70 mm  10 mm  0.010.1 mm), with a 1.525 kg
load cell. The speed was kept constant at 0.3 mm/s.
Nanoindentation was performed using an Agilent Nano
Indenter G200 (U9820A, Agilent Technologies, Inc.,
California, USA) at room temperature. A conical dia-
mond flat-punch indenter with a diameter of 50 nm was
13
12
11
10
9
Stress (MPa)
8
7 30% PCL
6 40% PCL
5 50% PCL
4 100% PCL
3
2
1
0
-1
0 5 10 15
Figure 1. Stress–strain curves of zein/PCL blend systems.
Textile Research Journal 88(24)
employed. Eight measurements were conducted per
sample as follows: a strain rate of 0.4/s was constantly
maintained during the addition of the load, until the
indenter reached a depth of 2000 nm on the surface of
the scaffold. The load was maintained at the maximum
value for 5 s. The indenter was then withdrawn from the
surface of the scaffold and unloaded to 10% of the max-
imum load. To ensure the reliability of these nanoscale
deformation measurements, at least five indents were per-
formed on each sample to obtain effective data.
Sample preparation. Zein/PCL blends with different
weight ratios were dissolved in N,N-dimethylforma-
mide, acetic acid and dichloromethane, The weight
ratios of the zein/PCL blends were 100/0, 90/10, 80/
20, 70/30, 60/40, 50/50 and 0/100. The solutions of
zein/PCL blends were magnetically stirred for 812 h
at room temperature. Zein/PCL (70/30) and its blends
containing different proportions of EPL were prepared
in the same way. Keeping the electrostatic spinning par-
ameters the same, the solutions were spun into fibrous
membranes. The applied voltage was fixed at
1520 kV; solution flow rate was 0.31 mL/h; needle
diameter was 1.0 mm; and the collecting distance was
610 cm. The concentration of the spinning solution
was 530%.
Antibacterial activity test
The preparation of liquid nutrient solution, trypticase
17 g, pepton 3 g, yeast extract 6 g, sodium chloride 5 g,
100% zein
10% PCL
20% PCL
20 25 30 35 40 45 50 55 60 65
Strain (%)
Hu et al. 2803

dipotassium phosphate 2.5 g, glucose 2.5 g and distilled
water 1000 mL were weighed. The above ingredients
were dissolved using heat mixing, adjusted to pH
7.27.4 and sterilized at 120 C for 30 min. The prepar-
ation of solid nutrient solution, trypticase 17 g, pepton
3 g, yeast extract 6 g, sodium chloride 5 g, dipotassium
phosphate 2.5 g, glucose 2.5 g, agar 15 g and distilled
water 1000 mL were weighted. The above ingredients
were dissolved using heat mixing, adjusted to pH
7.27.4 and sterilized at 120 C for 30 min.
The antibacterial activities of zein/PCL fiber mem-
branes with EPL (0%, 2%, 2.5%, 3%, 3.5%, 4%,
4.5%, 5%, 10% and 20%) were evaluated using the
zone inhibition method. L. monocytogenes cells were
used as a model microorganism. The microorganisms
were diluted in a liquid nutrient solution to obtain a cell
suspension containing 1  108 cells/mL and then culti-
vated in the water bath oscillator for 18 h at 37 C. The
bacterial suspensions were diluted, and the bacterial
suspensions of dilution were equably smeared on the
solid medium. All of the EPL-loaded zein/PCL fibrous
membranes were cut into 1.8 cm diameter pieces and
disinfected under UV light before testing. The 1.8 cm
diameter pieces were horizontally arranged on the solid
medium. The sealing cultivation box was placed in a
temperature incubator for 18 h at 37 C. The zone of
inhibition was determined by measuring the diameter of
the inhibition zone around each dish.
Results and discussion
Effect of zein content on tensile behavior
The mechanical properties of zein/PCL blends were
studied in order to determine the influence of PCL on
zein, as shown in Table 1. From the table, it is clear that
the tensile strength and elongation at break increased
linearly with the increase in PCL content. However, the
elastic modulus showed a slight decreasing trend.
Stress–strain curves of all the samples in Figure 1
showed ductile fracture. The deformation rate of zein/
PCL blend with 3050% PCL was larger than that of
zein/PCL with 1020% PCL. In fact, PCL could effect-
ively enhance the ductility and increase the deformation
rate of blends. The quadratic yield is a result of the
multilayer film structure within the blend systems.
Improvement in tensile properties, namely tensile
strength and elongation at break are very favorable
for applications such as facial masks, packaging and
wound healing. This improvement in mechanical prop-
erties could be attributed to the phase morphology of
the blends and the partial compatibility between the
two components.
In order to research the effect of PCL on the longi-
tudinal mechanical properties of zein, we tested the
mechanical properties of films by nanoindentation.
Nanoindentation tests have proven to be effective in
probing the mechanical properties of thin films and
composite materials at the micro- to nanoscale.25
Nanoindentation experiments performed on the fabri-
cated zein/PCL electrospun membranes demonstrated
ranges for elastic modulus (0.0850.163 GPa) and
hardness (0.0230.064 GPa), depending on the PCL
concentration. As shown in Table 2, with increasing
PCL (5060%) concentration, the elastic modulus of
the fibrous membranes increased from 0.118 GPa to
0.163 GPa. The elastic modulus of fibrous membranes
with PCL (30%) content decreased slightly. However,
the hardness of fibrous membranes increased with add-
ition of PCL. In Figure 2, the curves were shifted
upwards with increasing PCL content. This result
implied that the load applied on the indenter tip grad-
ually increased with increasing PCL content. From
Table 2 and Figure 2, it is evident that the elastic modu-
lus and hardness of zein improved significantly with
addition of PCL. This is an important property of
PCL as a structural material.
Dynamic mechanical analysis
The results of DMA are shown in Figure 3, which show
variations in tan  with temperature for the blend

Table 1. The mechanical properties of zein/PCL blend systems.

Zein/PCL Tensile Tensile Elongation Table 2. Comparison of elastic modulus and hardness of zein/
(wt/wt) strength (MPa) modulus (MPa) at break (%) PCL blend systems.

100/0 1.54 92 1.65 Zein/PCL Elastic Hardness

(wt/wt) modulus (GPa) (GPa)
90/10 1.58 37 4.33
80/20 2.03 44 4.76 100/0 0.089  0.009 0.023  0.003
70/30 2.32 21 11.88 70/30 0.085  0.023 0.030  0.008
60/40 3.79 22 17.34 60/40 0.118  0.013 0.049  0.013
50/50 5.18 17 27.5 50/50 0.163  0.017 0.032  0.002
0/100 12.36 28 49.29 0/100 0.152  0.074 0.064  0.031
2804 Textile Research Journal 88(24)

1.5
1.4 A 100% zein E
1.3
B 30% PCL
1.2
C 40% PCL
1.1 D
D 50% PCL
Load on Sample (mN)
1.0
0.9
E 100% PCL C
0.8
B
0.7
0.6
0.5
0.4
0.3
0.2
A
0.1
0.0
0 500 1000 1500 2000
Distance into Surface (nm)

Figure 2. Nanoindentation load–displacement curves of zein/PCL blend membranes.

A:PCL 100%
B:PCL 30%
C:PCL 40%
D:PCL 50% A
B
E:zein 100%
C
D

-80 -70 -60 -50 -40 -30 -20

A
B
C
D
E

-100 -80 -60 -40 -20 0 20 40 60 80 100 120 140 160 180 200
o
Temperature ( C)

Figure 3. Temperature dependence of tan  in zein/PCL blend systems: (a) 0/100; (b) 70/30; (c) 60/40; (d) 50/50; (e) 100/0.

systems. The two peaks of tan  as well as the Tg of the increased with increasing zein content in the blend sys-
blends can be seen. Tg of pure PCL was 74.31 C, tems. The values are presented in Table 3. This could be
whereas Tg of pure zein was 190.14 C. Tg of zein attributed to the difference in the crystallinity of PCL
remained almost unchanged, whereas Tg of PCL and zein. Since glass transition occurs due to transition
Hu et al.
Table 3. Comparison of DMA (tan ) for blend
systems.
Tg ( C)
Zein/PCL PCL Zein
0/100 74.31
70/30 25.40 193.10
60/40 38.07 193.88
50/50 52.05 193.68
100/0 190.14
from glassy to rubbery state in amorphous materials,
crystalline materials do not show this transition. The
Tg of blends is influenced more by the Tg of the more
amorphous phase, that is zein, and is less dependent on
the Tg of PCL, which is more crystalline and less
amorphous. With increasing zein content in blend sys-
tems, the peaks (Tg) of PCL shift inwards. This phenom-
enon confirms that the binary blend systems are partially
miscible biphasic systems. This may be due to certain
interactions between components of the blend. The par-
tially miscible biphasic systems help in improving the
mechanical properties of the blends.
Morphological analysis
Figure 4 shows the morphologies of ultra-fine fibers of
pure zein, pure PCL and zein/PCL (70/30), and blends
containing different proportions of EPL. All the samples
showed smooth morphologies with no beads, suggesting
that the polymeric concentrations of zein and PCL, the
choice of solvent system, as well as the electrospinning
parameters were all optimal. The fibers were well-
oriented. Morphologies of the oriented ultra-fine fibers
were extremely useful for the improvement of mechan-
ical properties. Meanwhile, the ultra-fine fibers of pure
zein (diameter 640 nm) were heterogeneous and coarser
than those of other systems, which could be attributed to
the lower viscosity of pure zein. The diameter of ultra-
fine fiber of PCL was 340 nm. With increase in PCL
proportion, the ultra-fine fibers of zein/PCL (70/30)
(diameter 460 nm) blends became more homogeneous
and much thinner. The reason for this phenomenon
was that the high viscosity of PCL could greatly increase
the viscosity of zein/PCL blends. The diameters of the
ultra-fine fibers of zein/PCL/EPL (24%) blends were in
the range 360220 nm, which were even smaller than
that of zein/PCL (70/30) blend. The diameters of ultra-
fine fibers of zein/PCL/EPL (4.520%) blends were in
the range 430530 nm, which were smaller than that of
pure zein, but larger than that of the zein/PCL (70/30)
blend. Generally, the diameter of zein fiber used in bio-
medical applications is in the range 400–500 nm).26
2805
Therefore, the ultra-fine fibers of zein/PCL blends pre-
pared in this study were also suitable candidates for bio-
logical applications. The diameters and standard
deviation (SD) of diameters of ultra-fine fibers are pre-
sented in Table 4.
Hydrophilicities of fiber films
The hydrophilicities of zein/PCL/EPL fibrous films
were evaluated by contact angle measurements, as
shown in Figure 5 and Table 4. It can be seen in
Figure 5(l) that the electrospun PCL film was a hydro-
phobic material. The contact angle of zein was 45.80 ,
whereas the contact angle of zein/PCL (70/30) was
69.30 . In addition, the fibrous films of zein/PCL
blend systems also exhibited hydrophobicity. It was
observed that the water drop could not be easily
dropped onto the surfaces of all zein/PCL systems,
although they did contact the surfaces of the films.
This suggested that they were hydrophobic. When
water droplets fell on the surface of film containing
zein, the contact angle rapidly decreased to almost
zero in 530 s. At the same time, it was also seen that
the contact angle decreased rapidly to almost zero with
increases in zein content. This marked change in con-
tact angle was probably due to the weak morphological
stability of zein in water that led to the collapse of the
spinning frame. On the contrary, the fibrous films of
PCL remained hydrophobic and showed strong mor-
phological stability against wetting. The contact
angles of zein/PCL/EPL (220%) blends were in the
range 64.10–39.00 . The hydrophilicity of the zein/PCL
blend increased with increasing EPL content. On one
hand, EPL was hydrophilic; on the other hand, EPL
had a low molecular weight. The hydrophilicity and
collapsing of the spinning frame increases with the
increase in EPL content. This phenomenon was related
to its chemical structure; the spinning of ultra-fine fibers
could resist and delay the penetration of water.
Antibacterial activity
The antibacterial activities of fibrous membranes of
electrospun zein/PCL (70/30), with and without EPL,
were assessed against typical pathogenic bacteria, such
as L. monocytogenes, as shown in Figure 6. From
Figure 6 it can be seen that there was no formation
of a bacteriostatic circle in absence of EPL. In contrast
to this, fibrous membranes of zein/PCL loaded with
EPL showed obvious inhibition zones for the two bac-
terial strains after 18 h intervals. Bacteriostatic circles
could be seen in samples with (220%) EPL content.
The lowest concentration of EPL in the blend that
showed a bacteriostatic circle was 2%. The bacterio-
static circle basically increases in size with increases in
2806 Textile Research Journal 88(24)

Figure 4. SEM micrographs of zein/PCL/EPL blend systems: (a) 100/0/0, (b) 0/100/0, (c) 70/30/0, (d) 70/30/2, (e) 70/30/2.5, (f) 70/30/
3, (g) 70/30/3.5, (h) 70/30/4, (i) 70/30/4.5, (j) 70/30/5, (k) 70/30/10, (l) 70/30/20.
Hu et al. 2807

Table 4. The diameters (mean) ultra-fine fibers and standard deviations (SD) of diameters of ultra-fine fibers, water contact angle
and water contact angle (SD) of fibrous membranes and the diameter of inhibition zones of fibrous membranes: zein/PCL/EPL (70/30/
020) blend systems

EPL (%) 0 2 2.5 3 3.5 4 4.5 5 10 20

Diameter mean (um) 0.46 0.36 0.31 0.26 0.23 0.43 0.49 0.50 0.51 0.53
Diameter mean (SD) 0.12 0.11 0.09 0.08 0.08 0.06 0.19 0.16 0.13 0.27
WCA 69.30 64.10 62.40 59.70 54.80 50.50 46.80 44.40 42.90 39.00
WCA (SD) 0.54 0.52 0.45 0.50 0.47 0.61 0.49 0.48 0.50 0.52
Zones (cm) 1.8 2.2 2.2 2.2 2.5 2.3 2.2 2.2 2.6 3.2

Figure 5. Water contact angle measurements of zein/PCL/EPL blend systems: (a) 100/0/0, (b) 70/30/0, (c) 70/30/2, (d) 70/30/2.5 (e)
70/30/3, (f) 70/30/3.5, (g) 70/30/4, (h) 70/30/4.5, (i) 70/30/5, (j) 70/30/10, (k) 70/30/20, (l) 0/100/0.
2808
Figure 6. The antibacterial activity of zein/PCL/EPL membranes against Listeria monocytogenes: (a), 70/30/0, 70/30/2, 70/30/2.5; (b)
70/30/3.5, 70/30/4, 70/30/4.5; (c) 70/30/3, 70/30/5, 70/30/10, 70/30/20.
EPL concentration. Moreover, the inhibition zone was
more remarkable for the fibrous membranes loaded
with a higher amount of EPL drug. These results indi-
cate that fibrous films of zein/PCL, loaded with EPL,
possessed good antibacterial properties and could be
used for wound healing, as facial masks and against
bacterial infections of the skin.
Conclusions
Mechanical properties of zein fibrous film could be
greatly improved by incorporating PCL. Zein/PCL sys-
tems were characterized by DMA, WCA, SEM, etc.
The ultra-fine fibers of zein/PCL systems had hydro-
philic surfaces, but were water insoluble. The PCL
ultra-fine fibers exhibited the lowest average diameter
of 336  90 nm. Increase in the amount of PCL in zein/
PCL systems resulted in reduction of the average diam-
eters of the ultra-fine fibers. These results indicate that
Textile Research Journal 88(24)
fibrous films of electrospun zein/PCL, loaded with
EPL, have good antibacterial properties. Results show
that the ultra-fine fibers of zein/PCL systems can
be conveniently prepared via electrospinning and can
be potentially used as tissue engineering scaffolds,
wound healing materials, food packaging and facial
masks. They can also be used for sustained release of
drugs, delivery systems of bioactive molecules and in
other biomedical applications.
Declaration of conflicting interests
The authors declared no potential conflicts of interest with respect
to the research, authorship and/or publication of this article.
Funding
The authors disclosed receipt of the following financial
support for the research, authorship, and/or publication
of this article: This study was financially supported by Jilin
Provincial Science and Technology Department (201215013),
Hu et al.
and was financially supported by the 2016 Training Programs
of Innovation and Entrepreneurship for Undergraduates.
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