materials

Article
Characterization of Antimicrobial Poly
(Lactic Acid)/Nano-Composite Films with Silver and
Zinc Oxide Nanoparticles
Zhuangzhuang Chu 1 , Tianrui Zhao 1 , Lin Li 2 , Jian Fan 1, * and Yuyue Qin 1, *
1 Institute of Yunnan Food Safety, Kunming University of Science and Technology, Kunming 650550, China;
18487262163@163.com (Z.C.); food363@163.com (T.Z.)
2 College of Light Industry and Food Science, South China University of Technology, Guangzhou 510640,
China; felinli@scut.edu.cn
* Correspondence: fanj333@163.com (J.F.); rabbqy@163.com (Y.Q.);
Tel.: +86-871-659-20216 (J.F.); +86-138-8819-5681 (Y.Q.)

Received: 8 May 2017; Accepted: 14 June 2017; Published: 16 June 2017

Abstract: Antimicrobial active films based on poly (lactic acid) (PLA) were prepared with nano-silver
(nano-Ag) and nano-zinc oxide (nano-ZnO) using a solvent volatilizing method. The films
were characterized for mechanical, structural, thermal, physical and antimicrobial properties.
Scanning electron microscopy (SEM) images characterized the fracture morphology of the films
with different contents of nano-Ag and nano-ZnO. The addition of nanoparticles into the pure
PLA film decreased the tensile strength and elasticity modulus and increased the elongation of
breaks—in other words, the flexibility and extensibility of these composites improved. According
to the results of differential scanning calorimetry (DSC), the glass transition temperature of the
PLA nano-composite films decreased, and the crystallinity of these films increased; a similar result
was apparent from X-ray diffraction (XRD) analysis. The water vapor permeability (WVP) and
opacity of the PLA nano-composite films augmented compared with pure PLA film. Incorporation of
nanoparticles to the PLA films significantly improved the antimicrobial activity to inhibit the growth
of Escherichia coli. The results indicated that PLA films with nanoparticles could be considered a
potential environmental-friendly packaging material.

Keywords: poly (lactic acid); nanoparticle; thermal properties; mechanical property; antimicrobial
property

1. Introduction
In last few decades, plastic bags made of polyethylene (PE), low-density polyethylene (LDPE),
high-density polyethylene (HDPE), and polypropylene (PP) have become common in daily life [1].
These polymers are non-biodegradable and use petroleum as their raw material, and the environment
can be contaminated with these bags [2]. Therefore, biodegradable and nontoxic packing materials have
a huge expanding area on the market. Poly (lactic acid) (PLA) is a biodegradable polymer which can
be produced from the bacterial fermentation of renewable resources, like sugar beet or corn starch [3,4].
PLA has been approved by the United States Food and Drug Administration (FDA). In addition, PLA
has other advantages, like moderate mechanical properties, transparency, commercial availability and
low price [5–8]. As a consequence, these advantages give PLA the best potential for packaging and
medical applications [6,8]. However, some properties of PLA, such as flexural properties, gas and
water vapor permeability and thermal stability, are not sufficient for certain applications [9]. In recent
years, many studies have been reported on improving some properties of PLA by blending it with
diversified additives, like plasticizer, other polymers and nanoparticles [9–12]. In our previous study,
we tried to prepare PLA composite films with nano-silver (nano-Ag) and nano-zinc oxide (nano-ZnO).

Materials 2017, 10, 659; doi:10.3390/ma10060659 www.mdpi.com/journal/materials

Materials 2017, 10, 659 2 of 13

Ag has2017,
Materials been10, known
659 to have antibacterial effects since ancient times [13], and recently, 2 ofAg
13 has
been used to control bacterial growth in many areas of applications [14–16]. Ag ions, even a low
plasticizer, other polymers and nanoparticles [9–12]. In our previous study, we tried to prepare PLA
concentration, have a very broad range of activity against microorganisms like bacteria, yeast, fungi,
composite films with nano-silver (nano-Ag) and nano-zinc oxide (nano-ZnO).
and viruses [17–20], and low toxicity to human cells [21]. Nevertheless, the synthesis of nano-Ag with
Ag has been known to have antibacterial effects since ancient times [13], and recently, Ag has
reduction in itstosize,
been used affects
control its chemical
bacterial growth and antibacterial
in many properties [14–16].
areas of applications considerably
Ag ions,[22].
evenToasolve
low this
problem, nano-Aghave
concentration, can be blended
a very broad with
rangepolymers like PLA
of activity against to form nano-composites
microorganisms [9], or
like bacteria, yeast, blended
fungi,
with and
a layer of metal-oxide, such as zinc oxide [23] and titanium dioxide [24],
viruses [17–20], and low toxicity to human cells [21]. Nevertheless, the synthesis of nano-Ag to form core-shell
morphologies that in
with reduction offer largeaffects
its size, surface area to volume
its chemical ratio. properties considerably [22]. To solve
and antibacterial
this problem,
ZnO nano-Ag
nanoparticles can the
have be blended
advantage withofpolymers
low cost,like
easyPLA to form nano-composites
availability, [9], or and
antibacterial effects,
blended
intensive with a layer
ultraviolet of metal-oxide,
absorption such as zinc
[25]. Moreover, oxide [23]
nano-ZnO hasandbeen titanium dioxide
approved [24],FDA
by the to form
as a safe
core-shell morphologies that offer large surface area to volume ratio.
material [23]. Hence, as a functional inorganic important material, nano-ZnO is increasingly being
ZnO nanoparticles have the advantage of low cost, easy availability, antibacterial effects, and
developed for use in research and health-related applications [26,27].
intensive ultraviolet absorption [25]. Moreover, nano-ZnO has been approved by the FDA as a safe
In this work, antimicrobial and degradable PLA nano-composite films with different contents
material [23]. Hence, as a functional inorganic important material, nano-ZnO is increasingly being
of nanoparticles
developed forwere use inprepared using
research and a solvent volatilizing
health-related applicationsmethod.
[26,27]. The objective of this study was
to evaluate the effects of these blend films on thermal, morphological,
In this work, antimicrobial and degradable PLA nano-composite films mechanical, gascontents
with different barrier, and
antimicrobial properties.
of nanoparticles Meanwhile,
were prepared usingthe structure-property
a solvent relationship
volatilizing method. for PLA
The objective nano-composite
of this study was
filmsto
was also analyzed.
evaluate the effects of these blend films on thermal, morphological, mechanical, gas barrier, and
antimicrobial properties. Meanwhile, the structure-property relationship for PLA nano-composite
2. Results
films was also analyzed.

2.1. X-ray Diffraction

2. Results

The structures
2.1. X-ray of pure PLA and PLA nano-composite films were analyzed by X-ray diffraction
Diffraction
(XRD) to study the effect of nanoparticle content on the crystallinity of the PLA matrix. The XRD
The structures of pure PLA and PLA nano-composite films were analyzed by X-ray diffraction
patterns of PLA and PLA nano-composite films are shown in Figure 1a–e. All the films showed
(XRD) to study the effect of nanoparticle content on the crystallinity of the PLA matrix. The XRD
intensity with a broad maximum appearing at approximately 2θ = 16◦ , which confirmed that the
patterns of PLA and PLA nano-composite films are shown in Figure 1a–e. All the films showed
PLA intensity
had no polymorphic crystalline
with a broad maximum transition
appearing [28]. For PLA/Ag/ZnO
at approximately 2θ = 16°, which blend films (Figure
confirmed that the1a–c),
it was observed that with the increase in the nano-ZnO content (0, 1, and 3
PLA had no polymorphic crystalline transition [28]. For PLA/Ag/ZnO blend films (Figure 1a–c), wt %) the intensity
it of
diffraction peaks atthat
approximately 2θ = 31.8 ◦ ◦ ◦
was observed with the increase in the, 2θ = 34.4 and
nano-ZnO 2θ =(0,
content 36.3 increased,
1, and 3 wt %)andthe these diffraction
intensity of
angles were consistent
diffraction peaks atwith the standard
approximately 2θ properties
= 31.8°, 2θfor hexagonal
= 34.4° and 2θ ZnO crystals
= 36.3° (Joint Committee
increased, and these for
Powderdiffraction angles
Diffraction were (JCPDS)
Studies consistentNo.with the standard
36-1451) properties for when
[29]. Furthermore, hexagonal ZnO crystals
compared (Joint
with a PLA/ZnO
Committee for Powder Diffraction Studies (JCPDS) No. 36-1451) [29]. Furthermore,
film (Figure 1d), the above diffraction peaks reduced or disappeared. This might have been due to when compared
with a PLA/ZnO
the decline of nano-ZnOfilm (Figure
content1d), theIn
[12]. above diffraction
addition, peaks reduced
the diffraction peaks or disappeared.
(Figure 1a–c) This might
evidenced at
have◦been due to the ◦decline of nano-ZnO content [12]. In addition, the diffraction peaks (Figure 1a–c)
2θ = 38.3 and 2θ = 44.4 fit the standard properties for FCC silver (JCPDS, No. 04-0783) [30], and the
evidenced at 2θ = 38.3° and 2θ = 44.4° fit the standard properties for FCC silver (JCPDS, No. 04-0783)
strength of these peaks were inconsistent. This could have been due to the addition of nano-ZnO with
[30], and the strength of these peaks were inconsistent. This could have been due to the addition of
different contents
nano-ZnO with[23].
different contents [23].

a
Intensity (a.u.)

10 20 30 40 50 60
2θ

Figure 1. The X-ray diffraction (XRD) patterns of: (a) Poly (lactic acid) (PLA)/Ag/ZnO-3;
Figure 1. The X-ray diffraction (XRD) patterns of: (a) Poly (lactic acid) (PLA)/Ag/ZnO-3;
(b) PLA/Ag/ZnO-1; (c) PLA/Ag; (d) PLA/ZnO; and (e) PLA.
(b) PLA/Ag/ZnO-1; (c) PLA/Ag; (d) PLA/ZnO; and (e) PLA.
Materials 2017, 10, 659 3 of 13

2.2. Fourier Transform Infrared Spectra

Fourier transform
Materials 2017, 10, 659 infrared (FTIR) spectra were measured for each film to study their3chemical of 13
structures. The principle pure PLA absorption bands in the infrared range were summarized in Table 1.
2.2. Fourier(IR)
The infra-red Transform
spectraInfrared and 2977.9 cm−1 were assigned to the asymmetric and symmetric
Spectra
at 2996.5
CH stretching
Fourierregiontransformof the –CH3(FTIR)
infrared mode, respectively.
spectra were measuredThe C=O stretching
for each of thetheir
film to study ester group was
chemical
attributed as a broad and strong absorption band at 1747.4 cm −1 . The region between 1500 and
structures. The principle pure PLA absorption bands in the infrared range were summarized in Table 1.
cm−infra-red
1360 The 1 was characterized
(IR) spectra atby the and
2996.5 1452.3 cmcm
2977.9 −1 −1CH
3 band.
were assignedThetoCH
the deformation
asymmetric and and asymmetric
symmetric
bands CH stretching
appeared at region − 1
1382.9ofcmthe –CH3 mode,cm
and 1359.7 − 1 , respectively.
respectively. The C=O stretching
Between the −ester
of cm
1300 1 andgroup
1000wascm−1 , it
attributedtoasobserve
a broad the and C=O
strongstretching
absorptionmode
band at of1747.4 cm group
. The region cm−1500
between 1 andand 1360 cm−1
−1
was possible the ester at 1267.1 the asymmetric
C–O–C wasstretching
characterized modeby theat 1452.3
1180.4, cm1128.2,
−1 CH3 band. The CHcm
and 1080.1 deformation and asymmetric
−1 . In the region of 1000 cm bands appeared
−1 and 800 cm−1 ,
at 1382.9 cm−1 and − 1 1359.7 cm−1, respectively. Between 1300 cm−1 and 1000 cm−1, it was possible to
the band at 956.2 cm was attributed to the characteristic vibration−1of the helical backbone with CH3
observe the C=O stretching mode of the ester group at 1267.1 cm and the asymmetric C–O–C
rocking mode. Two bands related to the crystalline−1 and amorphous phases−1of PLA were−1 found at 867.1
stretching mode at 1180.4, 1128.2, and 1080.1 cm . In the region of 1000 cm and 800 cm , the band
and 753.5 cmcm−1 . The peak at 867.1 cm−1 could be assigned to the amorphous phase and the peak at
at 956.2 −1 was attributed to the characteristic vibration of the helical backbone with CH3 rocking

753.5mode. −
cm Two1 to the crystalline phase [8,31,32]. and amorphous phases of PLA were found at 867.1 and
bands related to the crystalline
FTIR
753.5 cmspectra
−1. Theof theatpure
peak 867.1PLA
cm and
−1 couldPLA nano-composite
be assigned films arephase
to the amorphous shown andin Figure
the peak at2a–e.
753.5As
cmshown
−1

in Figure
to the2,crystalline
the curves of PLA
phase nano-composite films (Figure 2a–d) were similar to the pure PLA film.
[8,31,32].
However,FTIR withspectra of the pure
the addition PLA and PLA nano-composite
of nanoparticles, the band, at 1747.4, films are shown
1452.3, in Figure
1180.4, and 2a–e.
1080.1Ascm−1 ,
shown in Figure 2, the curves of PLA nano-composite films (Figure 2a–d)
increased in intensity. This may have occurred because the nucleation rate and nucleation density of were similar to the pure
PLA PLA film. However,
was increased with theofaddition
by addition of nanoparticles, the band, at 1747.4, 1452.3, 1180.4, and
nanoparticles.
1080.1 cm−1, increased in intensity. This may have occurred because the nucleation rate and nucleation
density of PLA was increased by addition of nanoparticles.
Table 1. Peak band assignments of pure PLA and PLA nano-composite films spectrums.

Table 1. Peak band assignments of pure PLA and PLA nano-composite films spectrums.
Assignment Peak Position (cm−1 )
Assignment Peak Position (cm−1)
–CH– stretch 2996.5 (asym), 2977.9 (sym)
–CH– stretch 2996.5 (asym), 2977.9 (sym)
–C=O carbonyl stretch 1747.4
–C=O carbonyl stretch 1747.4
–CH3 bend 1452.3
–CH3 bend 1452.3
–CH– deformation symmetric and asymmetric bend 1382.9, 1359.7
–CH– deformation symmetric and asymmetric bend 1382.9, 1359.7
–C=O bend 1267.1
–C=O bend 1267.1
–C–O– stretch 1180.4, 1128.2, 1080.1
–C–O– stretch 1180.4, 1128.2, 1080.1
–OH bend 1041.5
–OH bend 1041.5
–CH3 rocking modes 956.2
–CH3 rocking modes 956.2
–C–C– stretch 867.1
–C–C– stretch 867.1

b
Transmittance

3600 3300 30002000 1800 1600 1400 1200 1000 800 600
-1
Wavenumber (cm )

Figure 2. The Fourier transform infrared (FTIR) spectra of: (a) PLA/Ag/ZnO-3; (b) PLA/Ag/ZnO-1;
Figure 2. The Fourier transform infrared (FTIR) spectra of: (a) PLA/Ag/ZnO-3; (b) PLA/Ag/ZnO-1;
(c) PLA/Ag; (d) PLA/ZnO; and (e) PLA.
(c) PLA/Ag; (d) PLA/ZnO; and (e) PLA.
Materials 2017, 10, 659 4 of 13

Materials 2017, 10, 659 4 of 13

2.3. Scanning Electron Microscopy
2.3. Scanning Electron Microscopy
Scanning electron microscopy (SEM) images of cross-sectional microstructures of the blended
films wereScanning
shown electron
in Figuremicroscopy
3a–e. From (SEM) images
Figure of could
3, we cross-sectional
see that microstructures
the cross-sectionof the blended
of the pure PLA
films were shown in Figure 3a–e. From Figure 3, we could see that the cross-section of
film was a stratified structure, and by adding the nanoparticles, the microstructures were changed.the pure PLA As
film was a stratified structure, and by adding the nanoparticles, the microstructures were changed.
shown in Figure 3a–c, the stratified structure gradually disappeared and some small voids appeared
As shown in Figure 3a–c, the stratified structure gradually disappeared and some small voids
on the fractured surface of PLA/Ag/ZnO blend films, and more voids formed with the increase of
appeared on the fractured surface of PLA/Ag/ZnO blend films, and more voids formed with the
the nano-ZnO
increase of content. Furthermore,
the nano-ZnO the higherthe
content. Furthermore, thehigher
content
the of nano-ZnO,
content the more
of nano-ZnO, whitewhite
the more particles
aggregated. These aggregates indicated that the incomplete dispersion/blend of the
particles aggregated. These aggregates indicated that the incomplete dispersion/blend of the nanoparticles at
different content in the bulk of all films.
nanoparticles at different content in the bulk of all films.

Figure 3. Scanning electron microscopy (SEM) micrographs of the fracture morphology of:
Figure 3. Scanning electron microscopy (SEM) micrographs of the fracture morphology of:
(a) PLA/Ag/ZnO-3; (b) PLA/Ag/ZnO-1; (c) PLA/Ag; (d) PLA/ZnO; and (e) PLA.
(a) PLA/Ag/ZnO-3; (b) PLA/Ag/ZnO-1; (c) PLA/Ag; (d) PLA/ZnO; and (e) PLA.
2.4. Mechanical Properties
2.4. Mechanical Properties
Mechanical parameters such as elasticity modulus (EM), tensile strength (TS), and elongation of
break (E) areparameters
Mechanical important factors
such as in elasticity
evaluatingmodulus
polymeric films.tensile
(EM), The mechanical properties
strength (TS), of PLA
and elongation of
break (E) are important factors in evaluating polymeric films. The mechanical properties ofE PLA
films containing nanoparticles are shown in Table 2. For the pure PLA film, EM, TS and were films
measured
containing to be 3118.8are
nanoparticles MPa, 47.8 MPa,
shown and2.
in Table 5.35%, respectively.
For the pure PLAFurthermore,
film, EM, TSfromand Table
E were2, measured
it could to
be seen that addition of nanoparticles reduced the EM and TS values of the films and increased the E
be 3118.8 MPa, 47.8 MPa, and 5.35%, respectively. Furthermore, from Table 2, it could be seen that
value of the films compared with pure PLA film. The purpose of blending nanoparticles with PLA
addition of nanoparticles reduced the EM and TS values of the films and increased the E value of the
was to improve the plastic elongation and lower the brittleness.
films compared
The EMwithand pure PLAoffilm.
TS values PLAThe purpose of blending
nano-composite nanoparticles
films slightly changed whenwithcompared
PLA wastotopure improve
the plastic elongation and lower the brittleness.
PLA film. From Table 2, the EM values of PLA/Ag/ZnO blend films decreased by approximately
The
9.84%,EM and TS
29.32%, andvalues
18.94%of PLAincorporation
upon nano-composite of 0, 1,films
and 3slightly changed when
wt % of nano-ZnO compared
content, respectively.to pure
PLA PLA/ZnO
film. From Table
film 2, theby
decreased EM values
about of PLA/Ag/ZnO
16.29%. For the TS values, blend filmsnano-composite
the PLA decreased byfilms
approximately
fell to
28.16
9.84%, ± 2.69and
29.32%, MPa18.94%
from 44.30
upon± incorporation
3.62 MPa. The PLA/Ag/ZnO-3,
of 0, 1, and 3 wtPLA/Ag/ZnO-1
% of nano-ZnO andcontent,
PLA/ZnO films
respectively.
causedfilm
PLA/ZnO a significant
decreased(p <by
0.05) reduction
about 16.29%.in the
ForEMtheandTS TS values.
values, theThis result
PLA may have been films
nano-composite due tofell to
28.16the addition
± 2.69 MPa of antimicrobial
from 44.30 ± 3.62agents,
MPa.which decreased the interactions
The PLA/Ag/ZnO-3, between PLA
PLA/Ag/ZnO-1 chains and films
and PLA/ZnO
enhanced the mobility of PLA chains [33]. However, a slight reduction in EM and TS values was
caused a significant (p < 0.05) reduction in the EM and TS values. This result may have been due to the
addition of antimicrobial agents, which decreased the interactions between PLA chains and enhanced
the mobility of PLA chains [33]. However, a slight reduction in EM and TS values was observed to
Materials 2017, 10, 659 5 of 13

PLA/Ag/ZnO-3 when compared to that which was incorporated with 1 wt % nano-ZnO. This may
have been due to the molecular weight diminished and low molecular products formed.

Table 2. The mechanical properties of pure PLA and PLA nano-composite films.

Elasticity Modulus
Sample Tensile Strength (TS) Elongation of Break (%)
(EM)
PLA/Ag/ZnO-3 2528.20 ± 223.54 bc 36.15 ± 5.55 c 8.40 ± 0.11 b
PLA/Ag/ZnO-1 2204.31 ± 297.81 c 28.16 ± 2.69 d 12.22 ± 0.12 a
PLA/Ag 2811.76 ± 167.26 ab 44.30 ± 3.62 ab 6.68 ± 0.10 d
PLA/ZnO 2610.64 ± 297.51 b 38.96 ± 8.19 bc 7.74 ± 0.17 c
PLA 3118.79 ± 333.39 a 47.78 ± 5.18 a 5.35 ± 0.06 e
Values followed by different superscript letters (a–e) in the same column were significantly different (p < 0.05),
where a is the highest value.

The values of E of PLA/Ag/ZnO blend films was improved by 24.86%, 128.41%, and 57.01%
upon incorporation of 0, 1, and 3 wt % of nano-ZnO content, respectively. PLA/ZnO film increased
about 44.67%. This increase in the E value may have occurred because some plasticizing effect caused
by the addition of nanoparticles to the polymer matrix resulting in the increase in ductile properties,
which would also result in changes in the crystallinity of polymer [34]. It could be seen that the
E values did not increase with the increase of content of nano-ZnO. This might be because that
nano-ZnO concentration exceeded the maximum concentration allowed for forming homogeneous
composites [35]. Although the value of E increased compare with pure PLA film, it still low. To
improve the mechanical properties in the further applications, the addition of plasticizer into PLA
nano-composite films can be supplementary considered.

2.5. Differential Scanning Calorimetry

Differential scanning calorimetry (DSC) was used to study the glass transition, melting and
crystallization of pure PLA and PLA nano-composite films. DSC heating thermograms of pure PLA
and PLA nano-composite films were shown in Figure 4. DSC thermal properties such as the glass
transition temperature (Tg ), melting temperature (Tm ) and crystallinity (Xc ) are summarized in Table 3.
The Tg values of PLA nano-composite films were very small, and approximately 3 ◦ C lower than
the pure PLA film. In addition, small endothermic peaks were displayed within this temperature
range for the pure PLA and PLA nano-composite films, this might have been due to the typical effect
of physical aging [36]. The Tc values slightly decreased with the addition of nanoparticles, whereas
they decreased by 5.8 ◦ C in PLA/ZnO. This may have been due to heterogeneous nucleation, and
nanoparticles in PLA acted as the heterogeneous nucleus and promoted the cold crystallization of PLA
chains at lower temperatures [37]. The obvious peak at one of the highest temperature was regarded
as the melting temperature (Tm ) of all the films. The Tm values of PLA nano-composite films were
lower than pure PLA films. As was shown in Figure 4, there were double melting peaks near the
Tm . This double peak could be explained as the result of annealing occurring during the DSC scans
whereby crystals had time to recrystallize a few degrees above the melting point and then remelt [37].
When added the nanoparticles, the double melting peaks became clearer. The lower Xc (7.1–18.8)
of pure PLA and PLA nano-composite films was shown Table 3. However, amorphous regions of
PLA, which were affected by degradation, decomposed faster with lower Xc [38]. The DSC heating
process was essentially an annealing process. The Xc could be improved by annealing according to
the requirements of the biodegradation rate and the mechanical properties for the applications. All of
these results suggested that the nanoparticles could promote crystallization of PLA, including cold
crystallization and re-crystallization.
Materials 2017, 10, 659 6 of 13
Materials 2017, 10, 659 6 of 13

Heat Flow
d

Endo e

20 40 60 80 100 120 140 160 180 200

Temperature (℃)

Figure 4. Differential scanning calorimetry (DSC) curves of: (a) PLA/Ag/ZnO-3; (b) PLA/Ag/ZnO-1;
Figure 4. Differential scanning calorimetry (DSC) curves of: (a) PLA/Ag/ZnO-3; (b) PLA/Ag/ZnO-1;
(c) PLA/Ag; (d) PLA/ZnO; and (e) PLA.
(c) PLA/Ag; (d) PLA/ZnO; and (e) PLA.
Table 3. Thermal characteristics of pure PLA and PLA nano-composite films.
Table 3. Thermal characteristics of pure PLA and PLA nano-composite films.
Glass
Crystallization Melting
Transition Crystallinity
Sample Glass Transition Crystallization
Temperature Melting
Temperature
Temperature XCrystallinity
c (%) Xc
Sample Temperature Tg Tc (°C)Tc
Temperature Tm (°C) Tm
Temperature
(%)
(◦ C) Tg (°C) (◦ C) (◦ C)
PLA/Ag/ZnO-3 59.3 110.5 169.7 16.2
PLA/Ag/ZnO-3 59.3 110.5 169.7 16.2
PLA/Ag/ZnO-1 60.9 112.6 167.8 18.8
PLA/Ag/ZnO-1 60.9 112.6 167.8 18.8
PLA/Ag PLA/Ag 60.4 60.4 111.7111.7 169.0
169.0 13.6 13.6
PLA/ZnOPLA/ZnO 60.2 60.2 107.5107.5 170.7
170.7 15.3 15.3
PLA PLA 63.6 63.6 113.3113.3 173.2
173.2 7.1 7.1

2.6. Water Vapor Permeability

2.6. Water Vapor Permeability
Water was a noticeable factor in deterioration reactions and microbial growth of food, so
Water was a noticeable factor in deterioration reactions and microbial growth of food, so minimize
minimize even avoid the moisture transfer between food and the surrounding atmosphere became
evenimportant.
avoid the moisture transfer
In other words, between
Water vapor food and the surrounding
permeability (WVP) was one atmosphere became
of the most important.
important
In other words,inWater
properties vapor permeability
food packaging applications.(WVP)
The WVP was one of
values forthe
puremost
PLAimportant properties in food
and PLA nano-composite
packaging applications.
films were The WVP
shown in Figure 5. Thevalues
WVP for purePLA
of pure PLAfilmand PLA
was 1.69nano-composite
× 10−14 kg m/m2 sfilmsPa. Thewere
WVP shown
of in
FigurePLA/Ag/ZnO
5. The WVPblend of purefilms increased
PLA film was 1.69 × 10−(p
significantly 14 <kg
0.05)
m/m 2 s Pa.
with the The
increase
WVPofofnano-ZnO
PLA/Ag/ZnO contentblend
filmsfrom 1 to 3 wt
increased %. This behavior
significantly may with
(p < 0.05) have the
beenincrease
due to the hydrophilicity
of nano-ZnO of nano-ZnO
content from 1 toimproving
3 wt %. This
the hydrophilic interaction of films. On the other hand, the morphological
behavior may have been due to the hydrophilicity of nano-ZnO improving the hydrophilic interaction structures of PLA blend
films changed. This could be verified by SEM analysis showing that many voids in PLA/Ag/ZnO
of films. On the other hand, the morphological structures of PLA blend films changed. This could be
blend films and allowed more water vapor transfer. Although there was no significance difference
verified by SEM analysis showing that many voids in PLA/Ag/ZnO blend films and allowed more
in WVP between pure PLA film and PLA/Ag film (p > 0.05), the WVP value of PLA/Ag film was
waterequal
vapor totransfer.
the pureAlthough
PLA film there
in thewas noofsignificance
limit measurement difference
error. The in WVP between
addition pure PLA film
of nanoparticles
and PLA/Ag
enhancedfilm (p > 0.05),and
the flexibility the decreased
WVP value theofbarrier
PLA/Ag film was
properties of equal
the PLA to the pure
films. PLA results
Similar film inhad
the limit
of measurement
been studiederror. The researchers.
by other addition of nanoparticles
The WVP values enhanced
of PLA the flexibility
blend and increased
films were decreasedby thethe
barrier
properties
additionof the PLAagents
of other films.like
Similar results
nanosilica had
[39] andbeen studied
nano-Ag [9].by
In other
previous researchers.
studies, theThe
highWVP
WVP values
of of
film led to a slight decline in relative humidity within the packaging, which
PLA blend films were increased by the addition of other agents like nanosilica [39] and nano-Ag [9]. would be beneficial for
postharvest
In previous vegetable
studies, and fruit
the high WVP quality.
of filmFurthermore, there decline
led to a slight was no condensate
in relativewater when within
humidity using the
materials
packaging, with would
which high WVP. This would
be beneficial forhelp to reduce microbial
postharvest vegetablegrowthand fruit andquality.
maintainFurthermore,
an acceptable there
appearance of postharvest mushrooms [40]. Therefore, the PLA nano-composite films could be
was no condensate water when using materials with high WVP. This would help to reduce microbial
potentially be used for food packaging applications.
growth and maintain an acceptable appearance of postharvest mushrooms [40]. Therefore, the PLA
nano-composite films could be potentially be used for food packaging applications.
Materials 2017, 10, 659 7 of 13
Materials 2017, 10, 659 7 of 13

Materials 2017, 10, 659 3.0 7 of 13

3.0
2.5 a a
a b a

·s·Pa) ·s·Pa)
2.5
2.0 c c
b
2
10 kg·m/m
2.0
1.5 c c
-14 2
10 kg·m/m
1.5
1.0
WVP(×WVP(×
-14

1.0
0.5

0.5
0.0
PLA/Ag/ZnO-3 PLA/Ag/ZnO-1 PLA/Ag PLA/ZnO PLA

0.0 permeability (WVP) analysis of pure PLA and PLA nano-composite films.
Figure 5. Water vapor
Figure 5. Water vapor permeability (WVP)
PLA/Ag/ZnO-3 analysis of
PLA/Ag/ZnO-1 pure PLA
PLA/Ag and PLA nano-composite
PLA/ZnO PLA films. Values
Values followed by different superscript letters (a–c) in the same column were significantly different
followed
Figureby5.different superscript
Water avapor letters (a–c) in the same column were significantly different (p < 0.05),
(p < 0.05), where is the permeability
highest value.(WVP) analysis of pure PLA and PLA nano-composite films.
where a is the highest value.
Values followed by different superscript letters (a–c) in the same column were significantly different
(p < 0.05), where a is the highest value.
2.7. Opacity
2.7. Opacity
To a large extent, the appearance of goods influences the consumers to purchase them, so the
2.7. Opacity
transparency of packaging
To a large extent, is important.
the appearance ofThegoods opacity of the pure
influences thePLA and PLA to
consumers nano-composite
purchase them, filmsso the
is To a in
shown large extent,
Figure 6. thetransparency
The appearance of of goods
PLA influences the consumers
nano-composite films was to purchase
lower than them,
the pure soPLAthe
transparency of packaging is important. The opacity of the pure PLA and PLA nano-composite films
transparency of packaging is important. The opacity of the pure PLA
film. The opacity values of the PLA/Ag/ZnO-3, PLA/Ag/ZnO-1 and PLA/ZnO blend films were and PLA nano-composite films
is shown in Figure 6. The transparency of PLA nano-composite films was lower than the pure PLA
is shown in Figure
significantly 6. The
(p < 0.05) transparency
higher than pure of PLA PLA nano-composite
and PLA/Ag films. films was lower
This may thanbecause
have been the pureofPLA the
film. film.
The The
opacity values
opacity of the
values of PLA/Ag/ZnO-3,
the PLA/Ag/ZnO-3, PLA/Ag/ZnO-1
PLA/Ag/ZnO-1 and and PLA/ZnO
PLA/ZnO blendblend
films films
were were
color and the content of nanoparticles. The decrease in film transparency with PLA-base films by
significantly
adding
(p <(p0.05)
significantly < 0.05)
the antimicrobial
higher
higher than
agentsthanhadpure
pure
also
PLA
PLA been
and
and PLA/Ag
PLA/Ag
reported
films.
films.
[34]. TheThis Thisappearance
may
visual
maybeen
have havebecause
been because
of pureofPLA the of
the color
color and
and the
the content
content of
of nanoparticles.
nanoparticles. The The decrease
decrease in in
film film transparency
transparency with
and PLA nano-composite films is shown in Figure 7. Statistically significant differences were found with
PLA-base PLA-base
films by films
by adding
addingthe
between the antimicrobial
antimicrobial
samples agentshad
agents
with nanoparticles, hadalso
also
but been
been reported
reported
differences [34].
[34].
in visual The The visual
visual
appearance appearance
appearance
were oftopure
of pure
not notable the PLA
PLA
and
and PLA
human PLA nano-composite
nano-composite
eye. This result films films
suggested is shown
is shown
that, inin Figure
forFigure 7. Statistically
7. Statistically
consumers, significant
there was significant differences were
differences
no visual influence found
and were
peoplefound
between
could
between seesamples
samplesitems with
with nanoparticles,
sufficiently throughbut
nanoparticles, but differences
films wheninin
the differences visual
these appearance
films
visual were
were applied
appearance not
were notable
to the
not to theto the
packaging
notable
human eye.
applications. This result suggested that, for consumers, there was no visual influence and people
human eye. This result suggested that, for consumers, there was no visual influence and people could
could see items sufficiently through the films when these films were applied to the packaging
see items sufficiently through the films when these films were applied to the packaging applications.
applications.
a
8
a
8
b
6

b
Opacity

4
c
Opacity

4
c
2
d d
2
d d
0
PLA/Ag/ZnO-3 PLA/Ag/ZnO-1 PLA/Ag PLA/ZnO PLA
0
Figure 6. Opacity analysis of pure PLA and PLA nano-composite films. Values followed by different
PLA/Ag/ZnO-3 PLA/Ag/ZnO-1 PLA/Ag PLA/ZnO PLA
superscript letters (a–d) in the same column were significantly different (p < 0.05), where a is the
Figure 6.
highest Opacity analysis of pure PLA and PLA nano-composite films. Values followed by different
value.
Figure 6. Opacity analysis of pure PLA and PLA nano-composite films. Values followed by different
superscript letters (a–d) in the same column were significantly different (p < 0.05), where a is the
superscript letters (a–d) in the same column were significantly different (p < 0.05), where a is the
highest value.
highest value.
Materials 2017, 10, 659 8 of 13
Materials 2017, 10, 659 8 of 13

Materials 2017, 10, 659 8 of 13

Figure 7. The visual appearance of: (a) PLA/Ag/ZnO-3; (b) PLA/Ag/ZnO-1; (c) PLA/Ag; (d) PLA/ZnO;
The
Figure 7. and visual appearance of: (a) PLA/Ag/ZnO-3; (b) PLA/Ag/ZnO-1; (c) PLA/Ag;
(e) PLA.
(d) PLA/ZnO; and (e) PLA.
2.8. Antimicrobial Activity
To evaluate
2.8. Antimicrobial Activitythe antimicrobial properties of nanoparticles nano-ZnO and nano-Ag, the PLA
nano-composite films was studied for inhibition of E. coli growth, which was chosen in this study as
To evaluate the antimicrobial
a representative of common harmful properties of nanoparticles
microorganisms occurring in food nano-ZnO
products. Theandresults
nano-Ag,
of the the PLA
antimicrobial
nano-composite Figurefilms test are
7. Thewas shown
visualstudied in Figure
appearancefor 8. After
inhibition
of: (a) 12 h
PLA/Ag/ZnO-3; of
of E.the experiment,
(b)coli the
growth, (c)
PLA/Ag/ZnO-1; control
which
PLA/Ag; film,
was the
chosenPLA
pure
(d) PLA/ZnO; in this study
film, and
had(e)
noPLA.
antimicrobial activity to inhibit the growth of E. coli. The Log10CFU/mL value for pure
as a representative of common harmful microorganisms occurring in food products. The results
PLA film was up to approximately 9.21 from 5 at 0 h of the experiment. On the contrary, the
of the antimicrobial
Log
2.8. 10 CFU/mLtest
Antimicrobial valuesareof shown
Activity in Figure 8. After
the PLA nano-composite films with 12 antimicrobial
h of the experiment,
nanoparticlesthe showncontrol
to be film, the
pure PLAsignificantly
film, hadlower no antimicrobial
than the pure PLA activity
(p < 0.05),toand
inhibit
the valuesthe2.02,
growth E. coli.
of and
2.14, 2.98 3.31 Log The Log10 CFU/mL
10CFU/mL
To evaluate the antimicrobial properties of nanoparticles nano-ZnO and nano-Ag, the PLA
value for represented
pure PLAthe
nano-composite
PLA/Ag/ZnO-3,
film
filmswaswasup to PLA/Ag/ZnO-1,
studied approximately PLA/Ag and
for inhibition of E. 9.21 from
coli growth,
PLA/ZnO
5 at 0was
which
films,
of respectively
h chosen
theinexperiment.at the
this study as On the
end of 12 h of the experiment. These inhibitions were presumably due to the addition of
contrary, the Log10 CFU/mL
a representative of commonvalues of themicroorganisms
harmful PLA nano-composite occurring infilms with antimicrobial
food products. The results ofnanoparticles
the
nanoparticles. These nanoparticles released the surface of films through the micro-voids, formed in
shown to beantimicrobial test are shownthan in Figure 8. AfterPLA
12 h of the experiment, the control film, the 2.14,
pure PLA
the PLA nano-composite films by the nanoparticles, and restrained the growth of E. coli under the and 3.31
significantly lower the pure (p < 0.05), and the values 2.02, 2.98
film, had no antimicrobial activity to inhibit the growth of E. coli. The Log10CFU/mL value for pure
Log10 CFU/mL
presentrepresented the PLA/Ag/ZnO-3,
experimental condition [41]. On the one hand, PLA/Ag/ZnO-1,
the Log10CFU/mL PLA/Ag value of theand PLA/AgPLA/ZnO
film films,
PLA film was up to approximately 9.21 from 5 at 0 h of the experiment. On the contrary, the
was
respectively significantly (p < 0.05) less than the PLA/ZnO film. This may have been due to the different
Logat the end of 12 h of the experiment. These inhibitions were presumably due to the
10CFU/mL values of the PLA nano-composite films with antimicrobial nanoparticles shown to be
antimicrobial mechanisms of nano-ZnO and nano-Ag, and the low-dose of nano-Ag could be against
addition ofsignificantly
nanoparticles.lower thanThese nanoparticles
the pure PLA (p < 0.05), released the surface
and the values 2.02, 2.14,of films
2.98 through
and 3.31 the micro-voids,
Log10CFU/mL
the bacteria [17]. On the other hand, the PLA/Ag/ZnO films with a nano-ZnO content from 1 to 3 wt %
represented
formed inwere the
the significantly PLA/Ag/ZnO-3,
PLA nano-composite PLA/Ag/ZnO-1, PLA/Ag and PLA/ZnO films, respectively at the of E. coli
lower than the films by the nanoparticles,
other nano-composite andThis
films (p < 0.05). restrained
may have been the through
growth
end of 12 h of the experiment. These inhibitions were presumably due to the addition of
under thethepresent
synergy experimental
between nano-ZnO condition and[41]. On the
nano-Ag one hand,
improving the the Log10 CFU/mL
antimicrobial activity value
of of the
nanoparticles. These nanoparticles released the surface of films through the micro-voids, formed in
PLA/Ag filmnano-composite
was films when
significantly (pthese
< nanoparticles
0.05) less than were
the added
PLA/ZnO to the PLA
film. films.
This Inmay
fact, the
have ability
been fordue to the
the PLA nano-composite films by the nanoparticles, and restrained the growth of E. coli under the
the nano-composite films to inhibit other microorganisms on various food products at various
different antimicrobial
present experimentalmechanisms
conditionof nano-ZnO
[41]. On the oneand hand, nano-Ag, and thevalue
the Log10CFU/mL low-dose of nano-Ag
of the PLA/Ag film could be
storage temperatures should be tested in the future.
against thewas significantly
bacteria [17]. (pOn < 0.05) less than
the other the PLA/ZnO
hand, film. This mayfilms
the PLA/Ag/ZnO have with
been due to the different
a nano-ZnO content from
antimicrobial mechanisms of nano-ZnO 10 and nano-Ag, and the low-dose of nano-Ag could be against
1 to 3 wt % were significantly lower than the other nano-composite a films (p < 0.05). This may have
the bacteria [17]. On the other hand, the PLA/Ag/ZnO films with a nano-ZnO content from 1 to 3 wt %
0h
been through
were the synergy
significantly between
lower than the
8
nano-ZnO and nano-Ag
other nano-composite
12 h films (pimproving
< 0.05). This maythehave
antimicrobial
been throughactivity of
nano-composite
the synergyfilms between
when these nanoparticles
nano-ZnO and nano-Ag were improving
added to the the PLA films. Inactivity
antimicrobial fact, the of ability for
nano-composite films when these nanoparticles were added to the PLA films. In fact, the ability for
Log10CFU/mL

the nano-composite films to inhibit other microorganisms on various food products at various storage
6

the nano-composite films to inhibit other microorganisms on various food products at various
temperatures should be tested in the future.
storage temperatures should be tested in the future.
4
c b
10 d d
2
a
0h
8 12 h
0
PLA/Ag/ZnO-3 PLA/Ag/ZnO-1 PLA/Ag PLA/ZnO PLA

Sample
Log10CFU/mL

Figure 8. Antimicrobial activity of pure PLA and PLA nano-composite films. Values followed by
4
different superscript letters (a–e) b
in the same column were significantly
c
different (p < 0.05), where a is
the highest value. d
d
2

0
PLA/Ag/ZnO-3 PLA/Ag/ZnO-1 PLA/Ag PLA/ZnO PLA

Sample

Figure 8. Antimicrobial activity of pure PLA and PLA nano-composite films. Values followed by
Antimicrobial
Figure 8. different activity
superscript of pure
letters (a–e) PLAcolumn
in the same and PLA nano-composite
were significantly differentfilms. Values
(p < 0.05), where afollowed
is by
different superscript letters (a–e) in the same column were significantly different (p < 0.05), where a is
the highest value.
the highest value.
Materials 2017, 10, 659 9 of 13

3. Materials and Methods

3.1. Materials and Chemicals

Poly (lactic acid) (PLA, Mw = 280 kDa, Mw/Mn = 1.98) was purchased from Natureworks LLC
(Blair, NE, USA). Nano-Ag was purchased from Jinda Nano Tech Co., Ltd (Xiamen, Fujian, China).
Nano-ZnO was purchased from Qingdao nakasen Zinc & Technology Co., Ltd (Qingdao, Shandong,
China). All other reagents and chemicals used in the study were analytical grade.

3.2. Preparation of Films

Pure PLA and PLA nano-composite films were prepared by the solvent volatilizing method.
Before preparation, the PLA was dried in a vacuum oven at 80 ◦ C for 12 h to eliminate the influence of
water. A total of 2 g PLA was dissolved in 50 mL dichloromethane and stirred for 12 h. Then, different
amounts of nano-Ag or nano-ZnO was added to the PLA dichloromethane solution and stirred by
a magnetic stirrer until all of the nanoparticles were completely mixed. Subsequently, the solutions
were cast onto polytetrafluoroethylene (PTFE) plates and dried in a vacuum oven at room temperature.
The PLA film with 3 wt % nano-ZnO was named as PLA/ZnO, and with 0.5 wt % nano-Ag was
named as PLA/Ag. Nano-ZnO was incorporated into PLA/Ag as 1 and 3 wt % loading named as
PLA/Ag/ZnO-1 and PLA/Ag/ZnO-3, respectively. The pure PLA film was used as control.

3.3. X-ray Diffraction

The structure of nano-composites was evaluated by XRD, via an X-ray diffractometer (D8 Advance,
Brucker, Karlsruhe, Germany) with Cu Kα radiation, at a voltage of 40 kV and an electricity of 40 mA.
The samples were scanned in the diffraction angle 2θ, with a scan speed of 2◦ /min at room temperature.

3.4. Fourier Transform Infrared Spectra

FTIR measurements were accomplished using an FTIR spectrometer (Nicolet-5700, Ametek, Inc.,
Shanghai, China) in the range of 400–4000 cm−1 at a resolution of 4 cm−1 .

3.5. Scanning Electron Microscopy

The cross-section morphology of the blend films was performed with a scanning electron
microscope (NOVA NanoSEM 450, FEI, Hillsboro, OR, USA). Before the SEM examination, the films
were submerged in liquid nitrogen and fractured. The fracture surfaces of the films were coated with a
thin conductive gold layer in 20 nm thick.

3.6. Mechanical Properties

TS, E, and EM were measured with a Universal tensile machine (CMT 4104, MTS Systems Co.,
Ltd., Shenzhen, China) following the American Society for Testing and Materials (ASTM) Standard
Method D 882-88. The initial grip separation was set at 100 mm and the crosshead speed was set at
50 mm/min. The mechanical tests were replicated five times for each type of film.

3.7. Differential Scanning Calorimetry

DSC measurements were performed with a TA Instruments (DSC 214, Netzsch, Selb, Germany)
under nitrogen atmosphere. Firstly, the sample was sealed in an aluminum crucible and heated from
20 ◦ C to 200 ◦ C at a heating rate of 10 ◦ C/min, and held for 5 min to eliminate the previous thermal
history. Then the sample was cooled at 10 ◦ C/min to 20 ◦ C. Subsequently, the sample was heated again
from 20 ◦ C to 200 ◦ C at a heating rate of 10 ◦ C/min. The second heating scan was used to evaluate
Materials 2017, 10, 659 10 of 13

melting temperature (Tm ) and crystallinity (Xc ) of the samples. The percentage of crystallinity (Xc ) for
sample was calculated according to the following formula [42].

∆Hm − ∆Hc
Xc (% ) = × 100, (1)
∆Hm
o ×w

where ∆Hm is the melting enthalpy (J/g) of PLA in the sample, ∆Hc is the cold crystallization enthalpy
(J/g) of PLA in the sample, ∆Hm o is the heat of fusion for completely crystalline PLA (93.7 J/g) [43],

and w is the weight fraction of PLA in the samples.

3.8. Water Vapor Permeability

Water Vapor Permeability (WVP) of films was determined gravimetrically according to the ASTM
E96-95 standard method [44]. Previously, the test films were placed in the environment at 25 ◦ C and
50% relative humidity (RH) for 48 h. Then, weighing bottles were filled with allochroic silicagel and
the test films were sealed on the bottles by using paraffin and rubber. The covered bottles were put in a
temperature and RH controlled chamber to equate the conditions of test. The weight loss of each bottle
was measured as a function of time for 12 h. The WVP of the film was calculated with the following
formula [45].
WVTR × L
WVP = , (2)
∆P
where WVTR is the water vapor transmission rate (g/m2 s) through the film, L is the average film
thickness (m), and ∆P is the water vapor pressure difference (Pa) between the two sides of the film.
This entire test was repeated in triplicate for each type of film.

3.9. Opacity
Opacity was determined by measuring the absorbance at 600 nm using a UV-Vis
spectrophotometer (T90, Beijing Purkinje General Instrument Co., Ltd., Beijing, China) [46]. Each
sample was cut into a rectangle section (0.7 cm × 1.5 cm) and directly placed in a spectrophotometer test
cell. An empty test cell was used as the reference. The film opacity was calculated as following formula.

Abs600
Opacity = , (3)
d
where Abs600 is the value of absorbance at 600 nm and d is the film thickness (mm). This test was
triplicated for each type of film.

3.10. Antimicrobial Activity

The liquid culture test was used to evaluate the antimicrobial activity of PLA nano-composite
films. In other words, this test measured the ability of nanoparticles to restrain the growth of the
food pathogenic bacteria Escherichia coli. E. coli was obtained Laboratory of Microbiology, Faculty of
Life Science and Technology, Kunming University of Science and Technology, Yunnan, China. E. coli
was activated by inoculation into Mueller-Hinton broth. The bacteria were incubated at 37 ◦ C for
18–24 h. A glass test tube containing a testing sample (0.18–0.20 g) was filled with 10 mL of broth, and
inoculated with 0.1 mL of an overnight culture of E. coli. The bacterial cultures concentration was
adjusted to 105 CFU/mL. And these test tubes were shaken at 150 rpm at 22 ◦ C. After 0 and 12 h of the
experiment, aliquots containing 1 mL of sample were serially diluted with sterile phosphate buffer
(pH = 7.4) and then plated onto petri dishes. All petri dishes were incubated at 37 ◦ C for 24 h and
colony-forming units (CFU) were counted [41]. The pure PLA film was used as control.
Materials 2017, 10, 659 11 of 13

3.11. Statistical Analysis

The date were represented as the mean ±standard deviations and analysis of variance (ANOVA)
using SPSS (SPSS Inc., version 19.0, Chicago, IL, USA). Duncan’s multiple range test method was
applied to compare means for each test, with the statistical significance was defined at a level of
p < 0.05.

4. Conclusions
The work was focused on the key-characteristics of pure PLA and PLA nano-composite films.
PLA nano-composite films were prepared with nano-ZnO and nano-Ag by solvent volatilizing method.
According to DSC and consistent with XRD, which were demonstrated that the pure PLA and PLA
nano-composite films were mostly amorphous. FTIR analysis found that there were strong interactions
between PLA and nanoparticles. SEM analysis showed that the incompatibility of PLA and the addition
of nanoparticles in blends influenceed the morphology. The addition of nanoparticles into the pure
PLA film decreased the TS and EM, and increased the elongation of breaks, that is, the flexibility and
extensibility of the films improved. WVP and opacity properties of films were higher by the addition
of nanoparticles. In this study, the PLA nano-composite films showed good antimicrobial activity
against E. coli, especial the PLA/Ag/ZnO-3 and PLA/Ag/ZnO-1. In brief, the PLA nano-composite
films might have good potential for the application of antimicrobial packaging. Further research to
improve the plasticity of the composite films by adding plasticizer or essential oils with plasticization
will have further applications for the food packaging field.

Acknowledgments: The study was financially supported by the National Natural Science Foundation of China
(No. 21576126).
Author Contributions: Jian Fan and Yuyue Qin conceived and designed the experiments; Zhuangzhuang
Chu performed the experiments; Lin Li and Zhuangzhuang Chu analyzed the data; Tianrui Zhao contributed
reagents/materials/analysis tools; Zhuangzhuang Chu wrote the paper.
Conflicts of Interest: The authors declare no conflict of interest.

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