Ppap 201300152

Full Paper
Enhanced Antibacterial Activity of Melt

Processed Poly(propylene) Ag and Cu
Nanocomposites by Argon Plasma Treatment
Beatriz L. Espan~ a-Sa

nchez, Carlos A. Avila-Orta,* Felipe Padilla-Vaca,
Marıa G. Neira-Vela zquez, Pablo Gonza lez-Morones,
A. Rodrıguez-Gonza
Jose lez, Ernesto Hernandez-Herna ndez,

Angeles Rangel-Serrano, Enrique Dıaz Barriga-C., Luis Yate, Ronald F. Ziolo
Argon surface plasma treatment (APT) of poly(propylene)/silver (PP/Ag) and poly(propylene)/

copper (PP/Cu) nanocomposite improves their antibacterial properties against pathogenic
bacteria. Dispersions of PP/Ag and PP/Cu nanocomposites were prepared by sonication
assisted melt mixing 0.05, 0.5, and 5 w/w% metal nanoparticle/polymer compositions, and
then melt casted to form films that were surface treated with argon plasma. Scanning electron
and atomic force microscopy showed that APT increase the
surface roughness and the exposed nanoparticles. Also,
APT produced implanted oxygen and nitrogen species as
demonstrated by XPS analysis and enhanced the surface
wettability. Antibacterial activity (AA) against S. aureus
and P. aeruginosa of the plasma treated NC films was
significantly increased in all conditions tested such as NP
loading and interaction time; copper NCs were more
effective than silver NCs. Surface activation of the PP/Ag
and PP/Cu NC films by APT is a viable technique to increase
the antibacterial activity of nanocomposites, an important
issue in medical and health care applications.
B. L. Espan~ a-Sa
nchez, Dr. C. A. Avila-Orta, M. G. Neira-Velazquez, 1. Introduction
P. Gonza lez-Morones, J. A. Rodrıguez-Gonza lez, E. Hernandez-
Herna ndez, E. D. Barriga-C., R. F. Ziolo The design and application of polymer composites incorpo-
Centro de Investigacion en Quımica Aplicada (CIQA), Blvd. Enrique rating nanoparticles (NPs) have provided significant benefits
Reyna Hermosillo 140, Saltillo, Coahuila 25294, Me xico in areas related to medicine and health care since the
E-mail: [email protected] incorporation of these nanoparticles can drastically change
Rangel-Serrano
F. Padilla-Vaca, A.
and improve the desired properties of these materials in
Departamento de Biologıa, Divisio n de Ciencias Naturales y
targeted applications.[1,2] The antibacterial properties of such
Exactas; Universidad de Guanajuato. Noria Alta s/n, Guanajuato,
Guanajuato 36050, Me xico NCs have been extensively studied with a primary objective
L. Yate to prevent attack of several microorganisms in the above
Centro de Investigacio n Cooperativa en Biomateriales (CIC fields. Silver (Ag) and copper (Cu) in the elemental or partially
biomaGUNE), Parque Tecnolo gico de San Sebastia n, Paseo oxidized state have shown high antibacterial activity when
Miramo n 182, Ed. Empresarial C 20009, San Sebastia n, Spain added to different polymer matrices.[3–6] The antibacterial
Plasma Process. Polym. 2014, 11, 353–365

ß 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim wileyonlinelibrary.com DOI: 10.1002/ppap.201300152 353
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B. L. Espan nchez et al.
properties and effectiveness of polymer nanocomposites as received. Nanoparticles of elemental silver (20–30 nm diameter
depend on, among other things, the type, composition and per supplier) and copper (25 nm average diameter per supplier)
nature of the polymer matrix,[7] the nanoparticle composi- were purchased from SkySpring Nanomaterials Inc. (Houston, TX);
the powders were subject to sonication in air to decrease
tion, size and shape[8] and the effective interfacial contact
agglomeration before use. For the antibacterial tests, strains of S.
between the pathogen and the material.[9]
aureus ATCC #6838 and P. aeruginosa ATCC #13388 were used as
In order to enhance the antibacterial activity of polymers
received from American Type Culture Collection. Both micro-
and their surfaces, different techniques, such as spray- organisms were grown in Luria Bertani broth (LB) purchased from
ing,[10] sputtering,[11] plasma deposition,[12] and layer-by- BD Bioxon, Mexico. Sterile phosphate buffer saline (PBS) with
layer deposition,[13] have been applied to deposit nano- Tween 80 at 1% was prepared for use in the antibacterial tests using
particles on the polymer surface. In many of the cases, the ASTM standard E-2180-079 (2012): Standard Test Method for
NPs are physisorbed rather than chemisorbed on the Determining the Activity of Incorporated Antibacterial Agent (s) in
surface of the polymer. The lack of chemical bonding Polymeric or Hydrophobic Materials.
between the nanoparticle and the polymer could result PP pellets were hand-mixed with the commercially sourced silver
in easy release of the NPs to the environment by erosion and copper NPs at 0.05, 0.5, and 5 w/w % metal–polymer. Particle
with subsequent or eventual reduction of the antibacterial dispersion was achieved in three steps: (i) Premixing: Each sample
was prepared in a Brabender1 blender. The polymer was kept
properties of the polymer. In the case of melt mixing as a
molten at 190 8C for a period of 2 min before adding the NPs and then
nanocomposite preparative technique, however, nano-
mixed at 60 rpm for 8 min at that temperature. (ii) Dispersion: The
particles may be embedded in the polymer especially in premixed samples were cut and added to a Brabender1 extruder
the case of compatible surfaces. Melt mixing has been having a coupled 20 kHz ultrasound system. Pelletized samples
considered as an option to produce polymer NCs having were prepared at 190–195 8C and 60 rpm using the conditions
enhanced mechanical properties as well as potential
reported by Avila-Orta et al.[22] (iii) Film preparation: Pellets of each
antibacterial properties.[6,14,15] In this case, however, even sample from step (ii) were placed in a Killion1 2 roll, thin film
the near-surface NPs remain tightly embedded in the extruder at 190 8C, which was set to a film rate of 2 m min1. The
polymer to afford slow diffusion to the surface.[16] nanocomposites were obtained as films with an average thickness
Nonetheless, there are reports of NC preparations were of 200 mm. The rubber roller side of two cm diameter circular films
ion diffusion occurs, albeit slowly, that show significant were used for all experiments and subsequent analyses.
antibacterial activity.[17,18] In this report, we explored a
different strategy to produce lower cost NCs for antibacte-
2.2. Plasma Treatment
rial applications using melt mixing followed by argon
plasma treatment (APT) to enhance surface NP exposure. NCs films were placed in a cylindrical glass plasma reactor with an
In the ideal case, the surfaces of the active nanoparticles average volume of 6 L. Plasma was obtained with a 13.56 MHz
need to be exposed to their pristine with the NPs remaining radiofrequency plasma generator (Advanced Energy, RF600A),
which was inductively coupled to the cylindrical reactor through a
firmly embedded or chemically bonded to the polymer
matching network. All NC films were treated during 1 h at a plasma
surface.[6,15] To achieve this goal, plasma technology has been
power of 50 W under argon plasma; the gas pressure was 0.3 mbar
used to treat thin layers of polymer nanocomposites near while the flux was constant at 18.4 sccm; the source of Ar gas
the surface to expose the nanoparticles.[19,20] Armentano and was 99.999% pure (Infra, Inc). A schematic of the plasma system
coworkers[21] recently showed, for example, that oxygen is presented in the supporting information document.
plasma treatment of PLGA/Ag nanocomposite films resulted
in decreased bacterial adhesion and bacterial growth.
We have applied this concept to prepare metal–polymer 2.3. Morphological Characterization of Nanoparticles
nanocomposites with high antibacterial characteristics by and Nanocomposites
exposing the polymer NCs film surfaces to argon plasma.
The metal nanoparticles and PP/Ag and PP/Cu nanocomposites
We report here on the preparation and characterization were characterized by scanning and transmission electron micros-
of nanocomposites of poly(propylene)/silver (PP/Ag) and copies (SEM and TEM). SEM was done on a Jeol JSM-7041F at an
poly(propylene)/copper (PP/Cu) and on their subsequent accelerating voltage of 20 kV. TEM analyses were done on a FEI-
antibacterial activation by APT against Staphylococcus TITAN at 200 kV. Wide angle X-ray diffraction (WAXD) was carried
aureus and Pseudomonas aeruginosa. out on a Siemens D5000 in reflection mode using Cu Ka radiation
at 35 kV and filament intensity of 25 mA, at rate of 0.3u per second
from 5 to 808 with an interval of 0.028 in 2u.
2. Experimental Section
2.1. Materials 2.4. Surface Characterization of Nanocomposites
Isotactic poly(propylene) (PP) with a molecular weight (Mw) of Surface topography of the NCs was evaluated using atomic force
190 000 g mol1 was purchased from Sigma–Aldrich USA and used microscopy (AFM) in the tapping mode at a scan rate of 0.5 Hz and

354 ß 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim DOI: 10.1002/ppap.201300152
Antibacterial Activity of PP/Ag and PP/Cu Nanocomposites by APT
an average oscillation between 50 and 500 Hz on a Veeco sample, 100 mL of washed solution was plated on LB dishes and
Dimension 3000. Root mean square (RMS) roughness values were incubated for 16 h at 37 8C. Antibacterial activity percentage was
obtained in each analysis. Chemical changes at the surface of the calculated according to Equation (1),[23] where Co is the number of
films were determined using X-ray photoelectron spectroscopy bacterial colonies in the control film (pristine poly(propylene)) and
(XPS) with a model SPECS Sage HR 100 with a non-monochromatic C is the survival bacterial colonies on the NC films.
source. The selected resolution was 30 eV of Pass Energy and
0.5 eV step1 for the general survey spectra and 15 eV of Pass Energy Co C
Antimicrobial activity ð%Þ ¼ 100 ð1Þ
and 0.15 eV step1 for the detailed spectrum. Analyses were carried Co
out at a pressure of 5 108 mbar absolute with a takeoff angle of
908 using a magnesium source with Ka line of 1 253.6 eV of energy
and an applied power of 250 W. Gausian–Lorentzian functions 3. Results and Discussion
were used to perform the curve fitting of the high resolution
spectra. Changes in the wettability of nanocomposites were
In this study, APT of metal–polymer nanocomposite films of
investigated through static sessile drop contact angle measure-
PP/Ag and PP/Cu prepared by hot-roll thin film extrusion
ments using a Rame-Hart goniometer. Measurements were carried
out using droplets of 5 mL of deionized distilled water placed on the were investigated in order to (i) increase the ambient
surface of the samples, five measurements were performed for each surface exposure of metal nanoparticles, (ii) enhance the
sample to obtain an average of the contact angle. roughness and hydrophilicity of the film’s surface, and
(iii) increase the antibacterial characteristics of the metal–
polymer NC films and their effect on common pathogens
2.5. Antibacterial Tests as found, for example, in hospital environments.
Antibacterial properties of nanocomposite films were determined

using inoculums of S. aureus and P. aeruginosa grown in LB broth at
3.1. Characterization of As-Received Nanopowders
37 8C during 16 h. Untreated and plasma treated samples (P) were
sterilized using ultraviolet germicidal irradiation by exposition to HRTEM images, ED and EDX data for the as-received Ag and
254 nm for 30 min. Bacterial suspension was prepared in a sterile Cu NPs are presented in Figure 1a and b, respectively. ED and
solution of LB diluted five times with PBS, for a final concentration
XRD data confirm the composition as Ag and Cu with a thin
of 5 106 CFU mL1. 1.25 105 CFU mL1 was placed on the surface
coat of Cu2O on the latter. TEM analyses supported the
of the NC films and covered with a sterile cover slip. All samples
average particle sizes claimed by the supplier. Air sonica-
were incubated for 0, 90, 180, and 360 min at 37 8C. The assays were
carried out in duplicate in three independent experiments. The tion of the powders at 20 kHz helped attenuate agglomera-
bactericide effect was determined by washing each sample with tion during handling of the particles.
5 mL of PBS/Tween solution and shaking in an oscillating platform The commercial Ag and Cu NPs showed high anti-
for 5 min at 88 rpm to detach the bacteria from the film. From each bacterial characteristics and proved adequate for use in the
Figure 1. TEM characterization of as received nanoparticles before sonication: a) Ag NPs and b) Cu NPs. Upper left HR TEM image, Upper
right EDX, lower left ED and lower right histogram.

ß 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.plasma-polymers.org 355
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preparation of the desired metal–poly(propylene) NC agglomeration was more prevalent in the Ag NCs than in
precursor blends and films. As such, our approach circum- the Cu NCs, which gives the appearance of a higher number
vented the need to generate NPs either in situ or ex situ and density of particles for the Cu NCs in any given microtomed
to chemically functionalize them. The approach thus has section in the 5 wt% PP/Ag NC. In order to determine the
the added advantage of keeping the NPs in the pristine extent of the NP dispersion between that of the silver
state to afford direct metal bacteria contact. and copper in the 5 wt% NC’s, we used the method of
Khare and Burris,[24] which is based on measuring the
inter-nanoparticle distances from SEM our SEM data.
3.2. Polymer Nanocomposites: Morphology and
The dispersion for the Ag NCs has a homogeneity by an
Physicochemical Properties
average of 0.69 mm with the presence of some aggregates.
The nature of the dispersions achieved for the pre-film The Cu NCs show an average distance of 0.11 mm. These
materials prepared by the sonication assisted, melt blend/ results support the observation that the NPs in the Cu NCs
extrusion process (step ii) using the air sonicated Ag and Cu have a greater degree of dispersion than do those in the Ag
powders was deemed adequate for use in the preparation of NCs. This difference in the degree of dispersion is consistent
the final films used in this study. The average primary with the high copper exposition by plasma treatment and
particle size of the NPs after dispersion remained un- therefore high antibacterial properties.
changed as determined from SEM micrographs of many The difference observed between the particle sizes of
cryogenically microtomed NC samples, although some the Ag and Cu materials reflect the different surface
agglomeration was noted, particularly in the Ag NCs. chemistries of the Ag and Cu nanoparticles. The presence of
Figure 2 shows representative SEM micrographs of the the Cu2O coating on the Cu particles makes them less likely
type of agglomeration observed for the 5 wt% PP/Ag to agglomerate than the pure Ag particles and somewhat
and 5 wt% PP/Cu NC bulk blended materials. Some Ag relatively more compatible with the PP through possible
agglomerates with an average size of 100–150 nm may be hydrogen bonding in the solid state, particularly if the bulk
seen in Figure 2a while Figure 2b shows a representative PP was oxidized during thermal processing and sonication
dispersion of copper nanoparticles ranging in size from before film formation.[25]
about 30–60 nm, and some agglomerates. WAXD results (Figure 3a and b) show two characteristic
Good metal–polymer dispersions in the blends and films peaks at 14.28 and 21.48 2u, which correspond respectively
were achieved through sonication-assisted melt mixing of to the (110) and (131) planes of the a-mesophase for
the metal powders and commercially available isotactic PP isotactic PP.[26] This crystalline phase can form as a result
in two steps for the blends and an additional step for the of polymer and polymer composite orientation effects
films. The commercial NPs were air sonicated by an in- (independent of NPs) during film extrusion processing.[26,27]
house process just before melt mixing, which appeared to The sharp peaks in Figure 3a correspond to those for
aid in the subsequent dispersion of the NPs in the polymer. elemental silver, with attenuated intensity for NCs with
The nature of the bulk NC dispersions before the formation <5 wt% Ag.[28] The non-polymer peaks in Figure 3b for the
of the films was evaluated by SEM studies of cryogenically 5 wt% Cu NC at 42.98 and 36.58 2u are due to the Cu (111)
microtomed samples. The SEM images showed the Ag and and Cu2O (111) planes, respectively.[29] A small contribution
Cu NPs to be dispersed as primary particles and small from the cuprite Cu2O (200) planes (JCPDS 5-667) at 42.38 is
agglomerates (Figure 2) in all of the samples. In general, also possible.[30,31] The primary Cu0 peak can also be seen
Figure 2. SEM micrographs of cryogenically microtomed PP NCs after the melt blend and pellet extrusion process: a) PP/Ag-5 wt% and
b) PP/Cu-5 wt%.

Figure 3. WAXD data for PP NCs with different amount of nanoparticles: a) PP/Ag and b) PP/Cu.
in the 0.5 wt% sample. The presence of NPs in the NPs in the polymer surface in an average area of 28.28 mm2.
composites does not appear to modify the crystalline After plasma treatment (Figure 4b), some agglomerates
structure of poly(propylene). of Ag NPs with an average diameter around 100 nm
(average of three different images) can be seen due to
plasma polymer erosion and the subsequent exposure of
3.3. Surface Activation of the Plasma Treated
the Ag NPs. Similar behavior is observed in the Cu NCs,
Nanocomposites
where the untreated PP/Cu-5 wt% shows an average of
Representative SEM micrographs (secondary electron about three particles per 28.28 mm2 (Figure 4c) while that of
images) of untreated and plasma treated (P) 5 wt% PP/Ag the treated sample shows around 87 particles (Figure 4d),
are shown in Figure 4a and b, respectively. In the case of the again attributed to the plasma-induced polymer etching.
untreated films, Figure 4a does not show the presence of Ag The etching rate was evaluated based on the weight loss
Figure 4. SEM micrographs of a) untreated PP/Ag-5 wt%, b) plasma treated PP/Ag-5 wt%-P, c) Untreated PP/Cu-5 wt%, and d) plasma
treated PP/Cu-5 wt%-P.

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of the samples after the plasma treatment and was Table 1. RMS surface roughness (SR) of PP, PP/Ag-5 wt% and
approximately 1.8 nm min1. According to the optical PP/Cu-5 wt% treated and untreated with argon plasma.
emission spectra (OES) the etching processes was dominat-
ed by Ar ions and also by H ions. rms SR rms SR
Three-dimensional AFM images of the NCs were untreated plasma treated
obtained to evaluate the film roughness before and after Sample [nm] [nm]
APT. Figure 5a, c, and e (left column of Figure 5 images) show
PP 11.0 0.1 13.1 0.5
relatively smooth surfaces for untreated poly(propylene)
(PP), PP/Ag-5 wt% and PP/Cu-5 wt%, respectively. After PP/Ag-5 wt% 8.5 0.05 35.9 0.4
plasma treatment (PT), the topography of the nano- PP/Cu-5 wt% 17.6 0.05 109.4 0.1
composite films changes dramatically. The RMS value of
the surface roughness, SR, of the films before and after PT
is tabulated in Table 1. A slight increase is obtained in untreated sample. In the case of Cu NCs films, a sixfold
roughness of poly(propylene) film is shown in Figure 5b. increase was observed. In both cases, the number of
From Table 1, a fourfold increase in surface roughness of exposed NPs increases. On the other hand, AFM images
the silver nanocomposites films after PT as compared with provided in Figure 5d and f illustrate the increase in the
Figure 5. Three-dimensional AFM images of surface topography of untreated a) PP, c) PP/Ag-5 wt% and e) PP/Cu 5 wt%, and plasma treated
b) PP-P, d) PP/Ag-5 wt%-P and f) PP/Cu-5 wt%-P.

Table 2. Atom fraction from XPS analyses of untreated films of Cu NCs is of the same order for both AFM and SEM data.
PP, PP/Ag-5 wt% and PP/Cu-5 wt%, and plasma treated films of Moreover, the AFM images after PT clearly show the film
PP, PP/Ag-5 wt%-P and PP/Cu-5 wt%-P.
surfaces to be laden with the Ag and Cu NPs.
High resolution XPS spectra were used to elucidate the
Sample C N O Ag Cu chemical nature of the polymer NC films before and after
plasma treatment. XPS analyses were performed on films
PP 80.8 2.0 17.2 – –
stored in air for several months after film preparation
PP-plasma 76.0 6.0 17.9 – –
and PT. The general survey spectra of PP before and after
PP/Ag-5 wt% 94.6 0 5.4 0 – plasma treatment are shown in supporting information.
PP/Ag-5 wt%-P 90.2 0 9.6 0.2 – The atomic fraction of each element was obtained from
PP/Cu-5 wt% 97.2 0 2.5 – 0.3 the general survey spectra and they are reported in Table 2.
PP/Cu-5 wt%-P 76.5 4.2 16.7 – 2.7 As expected, analysis of the plasma treated PP, PP/Ag
5 wt% and PP/Cu 5 wt% all showed an increase in surface
nitrogen and oxygen, shown in Figure 6. The relatively
number density of exposed NPs after plasma treatment; high concentration of surface oxygen (22%) observed
as such, the images do not show the large four- and on the untreated PP film may be attributed to partial
sixfold increases, respectively, in surface roughness of oxidation of the tertiary carbon atoms in the PP repeat
the NC films (Table 1). Roughness may be increased units during thermal processing. Despite the possible
due to the resistance of the NPs to etching of the presence of these groups on the films before PT, and
plasma treatment, which is even in the polymer without no increase in the surface oxygen content of the plasma
NPs. The significant increase in surface roughness is treated PP films, the effect of the PT on the wettability of
presumably due to uneven erosion between areas of the films is strikingly significant as shown by contact angle
pristine polymer and the areas of the metal polymer measurements (vide infra).
domains and to localized effects caused by plasma The C1s spectra of the untreated PP/Ag-5 wt% and
interactions with the exposed, thermally conductive PP/Cu-5 wt% NC films are shown in Figure 7a and c,
metal particles and the surrounding polymer. Thus, for respectively, while the plasma treated sample spectra
the present NCs, surface roughness appears to result appear to their immediate right, Figure 7b and d,
from both chemical and physical changes. respectively. The fitted spectrum of PP/Ag-5 wt%-P after
The surface number density of exposed particles on plasma treatment (Figure 7b) shows a decrease in the
the treated and untreated films were counted using intensity of the 285 eV peak and a peak at 286.5 eV, which
the 2D AFM images (see Supporting Information) for an may be attributed to C—OH and C—N bond formation
average area of 900 mm2. PT of PP/Ag-5 wt% shows about during the plasma treatment and after exposure of the
100 particles compared with about 55 particles for the plasma treated films to air. The fitted spectrum of PP/Cu-
untreated film. The change for the PP/Cu-5 wt% film is 5 wt%-P after PT (Figure 7d) also shows the 285 eV (at lower
much more dramatic showing 650 particles after treatment intensity) and 286.5 eV peaks. The peak at 288.5 eV in the
versus 60 particles before treatment. The increase in copper NC could suggest the presence of C5 5O/O5 5C—OH
number density of exposed nanoparticles observed be- bonds. In both cases, plasma treatment of the PP nano-
tween the untreated and treated films for the Ag and composites followed by air exposure generates an expected
Figure 6. General survey spectra of a) untreated PP/Ag 5 wt% NC and plasma treated PP/Ag 5 wt% NC; b) untreated PP/Cu 5 wt% NC and
plasma treated PP/Cu 5 wt% NC.

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Figure 7. C1s spectra of untreated a) PP/Ag-5 wt%, c) PP/Cu-5 wt% and plasma treated b) PP/Ag-5 wt%-P, d) PP/Cu-5 wt%-P.
presence of oxygen and nitrogen species at the surface of were calculated to be 1.6 (0.3 atom fraction) and 12.1
the polymer (Table 2).[32] According to OES data obtained (2.7 atom fraction), respectively. The relatively high amount
during Ar plasma treatment of the films (not shown), only of surface Cu after plasma erosion (>5%) suggests a
Ar and H were detected. However, very low intensity redistribution of Cu particles toward the film’s surface
signals associated with N, O, and C ions were also detected. during the hot-roll melt extrusion processing of the films.
Thus, the increase in the N and O species after exposure to The higher amount of surface Cu relative to that of Ag as
air appears to come from both surface activation during determined by XPS is consistent with the above SEM and
the plasma treatment and reaction of the surface after AFM data.
plasma treatment upon exposure of the samples to air. The XPS, SEM, and AFM analyses clearly demonstrate
XPS Ag spectra of the untreated and plasma treated the difference in the nature of the NPs dispersions between
Ag NCs are shown in Figure 8a and b, respectively. For the pre-film, bulk blended materials and the final film-
the untreated Ag NCs, no peaks for Ag or its oxide were extruded materials used in our antibacterial study.
observed while for the plasma treated PP/Ag-5 wt%-P films Moreover, they also show the difference between the
(Figure 8b), two signals due to the Ag 3d5/2 and Ag 3d3/2 apparent amount of Ag and Cu on the films.
states appear at 368 and 375 eV, respectively.[33] The weight The wettability or hydrophilicity of the NC films
percent of surface Ag on the film was calculated to be 1.7 before and after PT as determined by contact angle
(0.2 atom fraction). The same behavior was observed for measurements are reported in Table 3. The contact angle
the 5 wt% copper NC films (Figure 8c and d, respectively) values of untreated films are for all films consistent with
with no Cu signal for the untreated films and a Cu 2p1/2 the hydrophobic surface of PP. After PT and air exposure, the
and Cu 2p3/2 peaks at 933.1 and 953.3 eV, respectively, films show hydrophilic behavior with contact angles of
for copper oxide, Cu2O[34] in the plasma treated films. about 608.
In addition, shake-up peak resonances for Cu2O appear The major physical and chemical attributes observed in
consistent with a surface coating of the oxide as observed the films after PT were (i) the substantial increase in the
in the ED and WAXD data above. The weight percentages number density of surface exposed NPs in the NC films and
of surface Cu on the untreated and plasma treated films (ii) the increased roughness (Table 1) and wettability of the

Figure 8. XPS of a) untreated PP/Ag-5 wt%, b) plasma treated PP/Ag-5 wt%-P, c) untreated PP/Cu-5 wt% and d) plasma treated
PP/Cu-5 wt%-P.
films (Table 3). The observed changes in the surface of the contact with pathogenic microorganisms suspended in
films before and after the plasma treatment may be the aqueous media.[38]
attributed to polymer erosion through the scission and loss
of polymer chains at the film surface due to argon ion
3.4. Antibacterial Activity
bombardment during the plasma treatment.[19,20] In
addition to the chain scission reactions, the collision of The antibacterial activity (AA) of the Ag and Cu NC films
the plasma-generated argon ions promotes the insertion before and after PT were evaluated by quantitative
of chemical species at the polymer surface[35,36] and the analyses of bacterial kill as a function of initial bulk NP
formation of active sites capable of reacting with air.[36,37] loading and bacteria-NCs contact times of 1, 3, and 6 h.
XPS analyses of the air-exposed, post plasma treated Figure 9a and b shows the AA, meaning the percentage
samples confirmed the presence of oxygen and nitrogen bacteria killed, for both the Ag and Cu NCs in contact with
species. Additionally, the resulting increase in the surface Staphylococcus aureus. The AA for the Ag and Cu NC films
adhesion changes of the NC films promoted more effective (Figure 9a and b, respectively) increased after PT and
contact time, with the 5 wt% Ag and Cu films showing
Table 3. Contact angle values of PP, PP/Ag-5 wt%, and PP/Cu-5 wt% up to 80 and 95%, respectively of bacteria killed after 6 h
nanocomposites. of contact. Because of the large difference in number
density of exposed NPs between the 5 wt% Ag and Cu NC
Sample Untreated Plasma treated films, no conclusions can be drawn about effectiveness
between the AA of one metal over the other except to say
PP 97 38 54 68 that the greater activity observed for the PP/Cu films
PP/Ag-5 wt% 100 28 648 48 may correlate in large part with the larger number density
PP/Cu-5 wt% 99 38 648 38 of exposed Cu NPs. However, comparing the AA of Cu

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Figure 9. Antibacterial activity of a) PP/Ag and b) PP/Cu nanocomposites untreated (solid bars) and plasma treated (hatched bars) in contact
with S. aureus.
and Ag commercial NPs used in this work, Cu NPs are more and with contact time. We observed that 100% of
effective to inhibit the S. aureus and P. aeruginosa growth P. aeruginosa are killed after about 180 min of interaction
than Ag NPs (data no shown). with PT Ag and Cu NCs at 5 wt%, except at 0.05 wt% loading
The physical and chemical changes generated by the APT where Cu NCs, unlike the Ag NCs, shows effective kill of
of the NC films significantly increased the antibacterial about 40% in 1 h.
activity of the PP/Ag[39] and PP/Cu NCs. In this study, the It is important to note that in both cases the anti-
gram-negative P. aeruginosa, was more susceptible to kill bacterial activity increased significantly with PT, where
in contact with the treated nanocomposites than was gram the Cu NPs exhibit higher antibacterial characteristics
positive S. aureus, which is consistent with the differences compared to those of the Ag NCs, which is attributable
in the nature of their cell walls.[40] Despite the apparent AA in large part to the higher number density of exposed
dependence on particle loading and time interaction, AA Cu NPs on the NC film surface. This notable increase in
was not observed in 0.05 and 0.5 wt% Ag NC films before activity for the present APT NCs is attributed to the
PT at any time tested, suggesting these films have not increase in nanoparticle exposure, surface roughness and
exposed NPs. This is supported by SEM and XPS results with hydrophilicity.
untreated 5 wt% Ag films where particles exposed are not In the case of the plasma untreated Ag NCs, the AA
detected and showed low AA against S. aureus. against the most susceptible bacteria does not depend of
The antibacterial activity of the Ag and Cu NC films direct contact with NPs, suggesting the release of Ag ions
against P. aeruginosa is shown in Figure 10a and b. The AA through the PP. The Ag ions are responsible of bacterial
for both the Ag and Cu films is much greater than that cell damage.[43] The results suggest that it is possible to
against S. aureus. This increase in activity most likely observe AA without direct contact of the NPS with the
relates to the differences in cell wall composition between bacteria, but that contact is critic to get an efficient AA.
the gram negative (P. aeruginosa) and gram positive The present plasma treated Ag NCs show faster bacterial
(S. aureus) bacteria.[41,42] The data for the untreated Ag kill at shorter contact times (3 and 6 h) as compared with
NCs (Figure 10a) show AA against P. aeruginosa despite earlier reports,[21,23,44,45] which commonly show anti-
NPs are not exposed. In general, AA increases after PT bacterial activity after a 24 h contact time.

Figure 10. Antibacterial activity of a) PP/Ag and b) PP/Cu nanocomposites untreated (solid bars) and plasma treated (hatched bars) in
contact with P. aeruginosa.
Fortunati et al.[21] determined the antibacterial activity melt mixing against E. coli and found that samples with
of PLGA/Ag NCs against S. aureus and E. coli prepared by 30% w/w of Cu NPs in PP inhibit growth of half of the
solvent casting with oxygen PT. They found that untreated original bacterial concentration after 1 h of interaction.
NCs with 7 wt% Ag showed approximately 20% of Delgado et al.,[6] have reported an AA of 95% for PP/Cu NCs
bacterial kill for 3 h of contact, and an increase to 25% with 5 vol% in contact to gram negative bacteria after 4 h
for 24 h of contact. Oxygen plasma treated samples show of interaction. Although Cu NPs were used in the initial
an AA up to 80% for both 3 and 24 h of contact with gram bulks samples, Cu and CuO were detected in the bulk by
positive and gram-negative bacteria. Other authors[23,44] XRD after thermal process. The authors reported no Cu
have studied the AA of Ag NCs obtained by plasma or CuO on the surface by XPS, and therefore concluded that
deposition. After 24 h of interaction with gram positive the antibacterial effect was by ion release. In our study,
and gram negative bacteria, the AA was found to be in the antibacterial behavior of the plasma treated Cu NCs
the range of 85 to 99% of bacteria kill. In our study, with increases significantly with 3 h of contact, where the NC
APT, Ag NCs with 5 wt% showed up to 80% of antibacterial with 0.05 wt% of copper shows an AA up to 90% after
effect at a low contact time (3 h) against S. aureus and contact with P. aeruginosa. Cu NCs in concentrations of 0.05,
100% with P. aeruginosa. Even at a contact time of 1 h, the 0.5, and 5 wt% with PT produce complete AA in contact
5 wt% NC films show better than 35 and 50% antibacterial with both pathogen microorganisms during 6 h of contact.
effect, respectively. These results are attributed to the APT We note that Cu, Cu2O, and CuO are all known to have
of the NC films, which serves to increase the number density antibacterial activity against S. aureus and P. aeruginosa.[46]
of NPs at the surface of NC films, and possibly promote or Although the antibacterial effect of Ag NPs is well
help in any subsequent initial leaching out of Ag ions. documented, few works compare the bactericidal effect
The copper nanocomposites show an important of Ag NPs versus that of the Cu NPs under the same
increase in their AA when Ar plasma treated compared conditions to assess which is more effective. The efficiency
to that of the untreated samples. Palza et al.[15] determined and magnitude of the antibacterial effect of NCs may
the antibacterial characteristics of PP NCs prepared by be attributed to several factors including ion release,

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Full Paper

Enhanced Antibacterial Activity of Melt

Beatriz L. Espan~ a-Sa

Argon surface plasma treatment (APT) of poly(propylene)/silver (PP/Ag) and poly(propylene)/

Plasma Process. Polym. 2014, 11, 353–365

Plasma Process. Polym. 2014, 11, 353–365

Antibacterial properties of nanocomposite films were determined

Plasma Process. Polym. 2014, 11, 353–365

Plasma Process. Polym. 2014, 11, 353–365

Plasma Process. Polym. 2014, 11, 353–365

Plasma Process. Polym. 2014, 11, 353–365

Plasma Process. Polym. 2014, 11, 353–365

Plasma Process. Polym. 2014, 11, 353–365

Plasma Process. Polym. 2014, 11, 353–365

Plasma Process. Polym. 2014, 11, 353–365

Plasma Process. Polym. 2014, 11, 353–365

Plasma Process. Polym. 2014, 11, 353–365

[32] M. Collaud, S. Nowak, O. M. Ku € ttel, P. Gro

Plasma Process. Polym. 2014, 11, 353–365

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