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DOI 10.1007/s13726-014-0258-3
ORIGINAL PAPER
Ali A. Katbab
Abstract Electrospun nanofibrous membranes (ENMs) Keywords Chitosan Poly(vinyl alcohol) Electrospun
were fabricated based on chitosan/poly(vinyl alcohol) (CS/ nanofibrous membrane Silver nanoparticle Antibacterial
PVA) with a 70/30 mass ratio containing silver nanopar- water filtration
ticles (AgNPs) via the electrospinning method. AgNPs
were produced on the surface of CS/PVA nanofibers by
adding AgNO3 to a CS/PVA blend solution as a silver Introduction
rendering component. The presence of AgNPs in the
polymer blend solution was detected by UV spectropho- Conventional water purification and disinfection methods,
tometry. The morphology of nanofibers before and after like chemical treatment and filtration by conventional
cross-linking with glutaraldehyde was investigated by the membranes, are not without problems. Chemical treatment,
field emission scanning electron microscopy. The forma- defined as the addition of biocides like chlorine-based
tion and size distribution of AgNPs onto the surface of components, leads to the production of carcinogen disin-
nanofibers were observed by transmission electron fection by-products (DBPs) [1]. In contrast, filtration of
microscopy and confirmed by energy dispersing X-ray microorganisms by conventional membranes leads to the
spectroscopy. As-spun and cross-linked CS/PVA nanofi- formation of biofilms, which significantly decrease the
bers revealed a smooth surface with diameters ranging quality of filtered water [2]. These problems in conven-
from 58 to 73 nm and 95 to 109 nm, respectively. The tional water purification and filtration systems make the
effect of AgNP formation on the chemical structure of improvement of water filtration systems an attractive
nanofibers was explored by Fourier transform infrared research trend. Electrospinning has been known as a simple
spectroscopy. Static and dynamic antibacterial filtration and effective method in producing continuous non-woven
efficiencies of CS/PVA ENMs, containing differing fibers with nanometric diameters [3, 4]. Electrospun
amounts of AgNO3, have been tested against Escherichia nanofibers, as a result of unique characteristics like high
coli, a gram negative bacterium. The antibacterial assess- surface area to volume, high aspect ratio, random orienta-
ment results exhibited a significant increase in both static tion, high air and water permeability; have been widely
and dynamic antibacterial filtration efficiencies of the studied for different applications including tissue engi-
prepared CS/PVA ENMs by addition of AgNO3 as a bac- neering, drug delivery, and filtration [57]. As recently,
tericidal agent. antibacterial water filtration using functionalized ENMs
has been investigateddue to high porosity, low pore size,
and high water fluxas an alternative to conventional
water purification and disinfection methods. Tan and
Obendorf [8] used nylon 6 ENMs to stabilize different N-
S. Adibzadeh S. Bazgir (&) A. A. Katbab halamines as bactericides to improve the antibacterial
Department of Polymer Engineering, Science and Research
activity of the prepared ENMs. The obtained ENMs com-
Branch, Islamic Azad University, PO Box: 14155-4933,
Tehran, Iran posite showed high antibacterial activity and low N-hal-
e-mail: bazgir@srbiau.ac.ir amine leaching. Daels et al. [9] worked on the application
of different biocides, like polychloride [(dimethylimino)(2- as CS/PVA solutions with 70/30 mass ratios, including
hydroxy-1,3-propanediyl)] (WSCP) and AgNPs, have been differing amounts of AgNO3. The prepared CS/PVA ENMs
tested to bond antibacterial properties to polyamide were then cross-linked using GA vapor as a cross-linking
nanofibers for the removal of gram negative and gram agent. The effect of the addition of AgNO3 on the chemical
positive bacteria. WSCPs containing polyamide nanofibers structure and morphology of the prepared nanofibers were
illustrate higher microorganism removal efficiency from observed using the Fourier transform infrared spectroscopy
water samples between different bactericides. In other (FTIR) and field emission scanning electron microscopy
works, CS-based ENMs were tried due to intrinsic anti- (FESEM) analyses, respectively. The formation of AgNPs
bacterial properties and biocompatibility, as next genera- in the polymer solution and onto the surface of CS/PVA
tion filters for antibacterial water filtration applications [10, nanofibers was illustrated by UV spectrophotometry and
11]. transmission electron microscopy (TEM), respectively. The
CS with a chemical structure of 1-4-linked-2-amino-2- presence of AgNPs on the surface of CS/PVA nanofibers
deoxy-b-D-glucopyranose is a biodegradable and biocom- was confirmed by energy dispersing X-ray spectroscopy
patible polycation acquired from chitin, the second most (EDX). The leaching behavior of AgNPs from CS/PVA
abundant polysaccharide after cellulose [12]. CS, because ENMs was determined using both static and dynamic
of the presence of amine groups in its backbones, shows methods. Static and dynamic antibacterial filtration effi-
inherent antibacterial activity, as well as metal binding ciencies of CS/PVA ENMs, with and without AgNPs,
properties in aqueous beds [13, 14]. Direct electrospinning against water samples containing E. coli, a gram negative
of CS is a challenging issue [15]. Internal hydrogen bacterium, have been evaluated. To our knowledge, this is
bonding between hydroxyl and amine groups in CS back- the first study which investigates different aspects of
bones is one of the main reasons for the electrospinning chitosan-based electrospun nanofibrous membranes con-
weakness of CS. The application of guest polymer with taining silver nanoparticles capability to be utilized in
good electrospinning ability, like PVA and PEO, is one antibacterial water filtration.
way to moderate the internal interaction between CS
backbones [1618]. In addition, solution properties such as
viscosity, ionic conductivity, surface tension, solvent type, Experimental
blend weight ratio, and polymer concentration are some of
the important factors which have direct effects on the Materials
spinnability of the CS solution [19]. Another way to
improve the spinnability of CS is to lower CS solution Medium molecular weight CS with a 7585 % degree of
surface tension by using concentrated acetic acid as a deacetylation (DDA) was purchased from Sigma Aldrich.
solvent. It is stated that, surface tension determines lower PVA (72,000 molecular weight), acetic acid (99.5 % gla-
and upper boundaries of the electrospinning window if cial), GA (25 % in water), and silver nitrate powder were
other factors are held constant. It is demonstrated that provided by Merck. Distilled water was used. All chemi-
although increasing the CS solution acidity leads to a slight cals were used as received without any further purification.
increase in solution viscosity, the CS solution surface
tension is decreased significantly [20, 21]. The CS can act Solution preparation and electrospinning
as a reducing agent to produce AgNPs from AgNO3. An
et al. [22] reported that the addition of AgNO3 to a CS/PEO CS solution was prepared by dissolving CS powder in
solution, as a silver rendering component, could produce 90 % acetic acid/water solvent under extensive stirring
AgNPs containing CS/PEO nanofibers after electrospin- conditions at 80 C for 4 h. PVA powder was dissolved in
ning, yielding higher antibacterial activity in comparison to double-distilled water under gentle stirring conditions at
plain CS/PEO nanofibers. Moreover, toxicity of the silver 80 C for 4 h in order to produce a homogeneous solution.
has been admitted to be significantly lower to human cells AgNO3 was added to the CS/PVA solution with a 70/30
than bacteria [23]. The Ag ion could interact with lone pair mass ratio to prepare blend solutions containing 0, 2, and 4
electrons of hydroxyl groups in PVA backbones, as well as wt% AgNO3 and a polymer concentration of 3 wt%. The
amine and hydroxyl groups in CS. These interactions alter obtained solutions were stirred for 5 h at 70 C to obtain a
Ag? to Ag, stabilizing Ag in the CS and PVA backbones homogeneous solution. The blend solutions were inserted
[24]. into a 60 mL syringe with a metallic needle (inner diameter
In the present study, CS/PVA ENMs, with and without gage of 18) that connected to a high voltage power supply.
AgNPs, were fabricated onto a polyester substrate by The voltage was adjusted to 18 kV and a solution flow rate
electrospinning technique. Ionic conductivity of CS/PVA was set to 0.1 mL/h using a syringe pump (AMPALL,
solutions was measured with different CS contents, as well Korea). A rotational steel collector which was covered by
Antibacterial assessments The CS/PVA nanofibers with 70/30 mass ratios containing
AgNPs were fabricated onto the polyester substrate to
The antibacterial properties of CS/PVA ENMs, with and improve the mechanical characteristics of the ENMs nee-
without AgNPs, were evaluated against PBS samples ded for the water filtration application. The effect of
varying CS contents in the CS/PVA solution on the ionic nanofibers under synergistic effect of viscosity and ionic
conductivity of the blend solution, as well as the effect of conductivity [27].
adding different amounts of AgNO3 into the CS/PVA In Fig. 3, the FESEM images and fiber size distributions
solution with a 70/30 mass ratio on the ionic conductivity of CS/PVA nanofibers with 70/30 mass ratios containing
of the CS/PVA solution is depicted in Fig. 1. As shown in 4 wt% AgNO3 before cross-linking (Fig. 3a, b) and after
Fig. 1 by increasing the CS/PVA ratio of the blend solu- cross-linking (Fig. 3c, d) are shown. It can be determined
tion, the ionic conductivity is decreased. It is reported that from Fig. 3 that the mean fiber diameter is shown to
the viscosity of the CS/PVA solution is lowered by increase from 59 10 to 96 12 nm after cross-linking.
increasing the CS content [26]. In addition, the addition of It is well-known that increases in mean fiber diameter,
AgNO3 into the blend solution leads to higher ionic con- along with white to yellow color changing, are character-
ductivity of the solution. istic of a cross-linking reaction between hydroxyl and
The low ionic conductivity of the CS/PVA solution with amine groups of CS with aldehyde groups of GA [28].
a 90/10 mass ratio led to the formation of nanofibers with
beaded structures (results not shown). By decreasing the Characterization of Ag nanoparticles
CS/PVA mass ratio from 90/10 to 70/30, the ionic con-
ductivity of the CS/PVA solution was increased from As functionalizing agent, the formation of AgNPs into the
860 8 to 964 10 ls/cm which facilitated the forma- blend solution and onto the surface of the CS/PVA
tion of jet polymer during the electrospinning process and nanofibers was confirmed by UV spectra of the CS/PVA
subsequently contributed in the formation of bead-free solutions containing different amounts of AgNO3 and by
fibers. The FESEM images of CS/PVA nanofibers with using EDX analyses, respectively (Fig. 4a, b). It can be
70/30 mass ratios containing 0, 2, and 4 wt% AgNO3 are observed from Fig. 4a that by adding AgNO3 into the blend
shown in Fig. 2. It can be seen from Fig. 2 that as-spun solution, a broad peak appears in its UV spectra around
nanofibers containing 0, 2, and 4 wt% AgNO3 show bead- 430 nm. The appearance of this peak is evidence of the
free structures with mean diameters of 73 15, 65 12 formation of AgNPs in the blend solution [29]. By
and 59 10 nm, respectively. A slight decrease in the increasing the AgNO3 content from 0 (Fig. 4c) to 2
mean fiber diameters of CS/PVA nanofibers containing (Fig. 4d) and to 4 wt% (Fig. 4e), the peak intensity is
AgNPs can be correlated with the higher ionic conductivity increased. This increase in AgNO3 causes an increase in
of the solution, derived from the presence of AgNO3 in the AgNPs content of the solution. The EDX analysis was
polymer solution, as presented in Fig. 1. Also, the addition used to confirm the presence of AgNPs on the surface of
of AgNO3 increases the viscosity of CS/PVA solution, nanofibers by detecting the appearance of a peak around
which could prevent fracturing of polymer jets that may 3 keV in the EDX graph, which is characteristic of the Ag
contribute in the formation of smooth and continuous element (Fig. 4b) [30].
Fig. 3 FESEM images of CS/PVA nanofibers containing 4 wt% AgNO3: a, b before cross-linking and c, d after cross-linking
In Fig. 5 the TEM images of CS/PVA nanofibers con- AgNPs are formed on the surface of nanofibers. This can be
taining 2 (Fig. 5a) and 4 wt% AgNO3 (Fig. 5b) are shown. explained by the presence of higher amine groups on the
It can be seen from Fig. 5 that by increasing the AgNO3 surface of nanofibers, where there is a higher CS content.
content from 2 to 4 wt%, the mean diameter of the particles This is in line with the observations of Penchev et al. [32].
increased from 10 4 to 34 10 nm without signifi- Phase separation of the CS and PVA in the CS/PVA
cantly increasing the number of AgNPs. In Fig. 5b, nanofibers is reported by Zhang et al. [19].
aggregation of the AgNPs into clusters on the fiber surface
can clearly be seen. It is found that AgNP formation starts FTIR analyses
with the nucleation of reduced Ag? ions into Ag atoms
followed by the growth of Ag nuclei. The nucleation of the The FTIR spectra of CS bulk, as-spun CS/PVA nanofibers,
AgNPs requires higher activation energy than the amount and CS/PVA nanofibers containing AgNPs are shown in
needed for the growth stage of AgNPs [29]. It is reported Fig. 6. From FTIR spectra of bulk CS it is evident that
that increases of the CS content in CS/PVA/Ag? solution absorption peaks at 1,599 and 1,661 cm-1 are attributed to
up to 12.5 wt% lead to lowering number of stabilized amide II and amide I, respectively. The CO stretching
AgNPs and an increase in particle size [31]. In this study, band is shown at 1,025 cm-1 and the CH stretching band
the CS content of the blend solution has been increased up is observed at 2,873 cm-1. An absorption peak at
to 70 wt%, which provides higher amine and hydroxyl 3,376 cm-1 is characteristic of the OH and NH
groups in the polymer solution that could interact with the stretching vibrations. FTIR spectra of CS/PVA nanofibers
Ag? ions by chelation and gathering together. These show that the addition of PVA to CS in CS/PVA nanofibers
attractions between Ag? and amine groups increase under would sharpen the absorption peaks of CS/PVA nanofibers,
the electrostatic force during electrospinning. Moreover, which allows for hydrogen bonding between OH groups
increasing the AgNO3 content in the solution provides of PVA with NH and OH groups of CS backbones [19].
more Ag? ions that could be enclosed by chelation. Ngu- An additional absorption peak at 1,727 cm-1 is charac-
yen et al. [31] have stated that large AgNPs are formed on teristics of C=O bands with acetic acid solvent remaining
the electrospun nanofibers by diffusion and aggregation of in the nanofibers. The comparison of FTIR spectra of as-
the residual amount of Ag ions after electrospinning during spun CS/PVA nanofibers, with and without AgNPs, dem-
the heat treatment. Furthermore, it is clearly shown that onstrates that the addition of AgNPs significantly lowers
the amine absorption peaks at 1,599 and 3,419 cm-1. stabilization of Ag by the formation of NAg groups
These changes in peak intensities of the amine groups through the chelation process.
illustrate the consumption of NH2 groups and the
Leaching kinetics
Fig. 7 Dynamic silver release test and static silver release test Fig. 8 Log of E. coli concentration in PBS samples after soaking
10 mg of nanofibers containing a 0, b 2, and c 4 wt% AgNO3
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