Materials Chemistry and Physics 134 (2012) 557e561

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Materials Chemistry and Physics

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Materials science communication

One-step fabrication of antibacterial (silver nanoparticles/poly(ethylene oxide)) e Polyurethane bicomponent hybrid nanobrous mat by dual-spinneret electrospinning
Leonard D. Tijing a, *, Michael Tom G. Ruelo a, Altangerel Amarjargal b, c, Hem Raj Pant b, d, Chan-Hee Park b, Cheol Sang Kim a, b, *
a

Division of Mechanical Design Engineering, Chonbuk National University, Jeonju, Jeonbuk 561-756, Republic of Korea Department of Bionanosystem Engineering, Chonbuk National University, Jeonju, Jeonbuk 561-756, Republic of Korea Power Engineering School, Mongolian University of Science and Technology, Ulaanbaatar, Mongolia d Department of Engineering Science and Humanities, Institute of Engineering, Pulchowk Campus, Tribhuvan University, Kathmandu, Nepal
b c

a r t i c l e i n f o
Article history: Received 26 January 2012 Received in revised form 9 March 2012 Accepted 22 March 2012

a b s t r a c t
The one-step electrospinning fabrication of novel materials with added functionalities is being widely studied because of their wide array of applications. Here, the fabrication of a hybrid, bimodal nanobrous mat made of two polymeric nanobers: polyurethane (PU) and silver (Ag) nanoparticle (NP) e in situ e decorated poly(ethylene oxide) (PEO) utilizing an angled dual-spinneret electrospinning system is reported. Silver nitrate (AgNO3) is in-situ reduced in high-molecular weight PEO, and Ag NPs with sizes from 6 to 90 nm as checked by scanning electron microscoy and transmission electron microscopy, are subsequently formed on the surface of PEO nanobers depending on the reduction time. Successful fabrication of bicomponent polymer matrices (PU and PEO) in the hybrid mat is conrmed by Fourier transform infrared spectroscopy. Metallic Ag NPs are veried to be present in the hybrid mats by energy dispersive X-ray spectroscopy and ultraviolet-vis spectroscopy, showing plasmon resonance band peaks at 415 and 425 nm. The hybrid nanobrous mat containing Ag NPs with an average size of 8 nm (i.e., reduction time of 3 h) exhibits strong antibacterial activity. 2012 Elsevier B.V. All rights reserved.

Keywords: Composite materials Electrospinning Nanostructures Polymers

1. Introduction The unique properties of non-woven polymer nanobers fabricated by electrospinning have evoked signicant interests in different elds [1,2]. Among the current studies, the electrospinning fabrication of hybrid materials by blending two or more polymers or the incorporation of metal nanoparticles (NPs) in/on polymer nanobers has gained vast attention [3e6]. Recently, multispinneret electrospinning presents itself as an excellent method to fabricate hybrid materials with improved properties by utilizing the different properties of each component polymer [7,8]. Many of the studies on multi-spinneret electrospinning focused on the effect of blended polymers on the improvement of hybrid materials overall properties [9e11]. To the authors best knowledge, no one has

* Corresponding authors. Division of Mechanical Design Engineering, Chonbuk National University, Jeonju, Jeonbuk 561-756, Republic of Korea. Fax: 82 632702460. E-mail addresses: ltijing@jbnu.ac.kr (L.D. Tijing), chskim@jbnu.ac.kr (C.S. Kim). 0254-0584/$ e see front matter 2012 Elsevier B.V. All rights reserved. doi:10.1016/j.matchemphys.2012.03.037

reported yet on the fabrication of hybrid nanober mat by dualspinneret electrospinning composed of mixed matrices of silver (Ag) NP e decorated poly(ethylene oxide) (PEO) and polyurethane (PU) nanobers. There are basically two ways to prepare metal NP-decorated polymer nanobers: (a) by synthesis of NPs and then dispersion on nanobers [12]; and (b) by in-situ reduction of metal NPs within or at the interface of nanobers [1]. The former is a two-step process, and presents difculty in achieving well-dispersed NPs. In this study, we present a one-step fabrication of hybrid nanober mat composed of two different polymeric nanobers (PEO and PU), wherein only PEO is decorated with Ag NPs without using any chemicals, i.e. PEO is used as both the reducing agent of silver nitrate (AgNO3) and the template of the formed Ag NPs. PU is one of the most used polymers in biomedical applications due to its good biocompatibility and excellent mechanical properties [13,14]. PEO is a non-toxic and biocompatible polymer that is widely used for tissue engineering, scaffolds, and wound dressing [15]. Silver NPs possess an inherent antibacterial activity against a number of pathogenic bacteria, have good biocompatibility [13], and are widely

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used as catalysts, and photosensitive components [16]. A recent review by Dallas et al. [17] discussed about the use of silver polymeric nanocomposites as advanced antimicrobial agents. Here, the combination of PU and Ag/PEO nanobers to make into a bicomponent and bimodal hybrid mat is potentially useful as antibacterial tissue scaffolds or for wound-healing applications. The bigger PU nanobers give structural stability and prolonged use of the hybrid materials [18], while the presence of thinner PEO nanobers provide more reactive surface area, and the decoration of Ag NPs on PEO surfaces without the use of chemicals gives antibacterial functionality to the hybrid mat. 2. Experimental section In a typical experiment, two separate solutions were prepared for dual-spinneret electrospinning. In solution A, 3.3 wt% poly(ethylene oxide) (PEO, molecular weight (MW) 60 kDa, Acros Organics) and 5 wt% silver nitrate (AgNO3, 99.8%, Showa) relative to the weight of PEO, were dissolved in distilled water (DW) in two separate glass containers. In one container, 2 g PEO was dissolved in 48 g DW, and in another container, 100 mg AgNO3 was dissolved in 10 g DW. After complete dissolution (around 4 h of magnetic stirring for PEO/DW solution), the two solutions were combined by magnetic stirring in covered containers and kept away from light for the reduction of AgNO3 into Ag NPs. The ratio of AgNO3 to PEO in the mixed solution was 1:20. In Solution B, 10 wt% of polyurethane (PU, Estane Skythane X595A-11, Lubrizol) was dissolved overnight in solvent mixture of N,N dimethylformamide (DMF, Showa) and methyl ethyl ketone/2-butanone (MEK, extra pure, Junsei) (50/ 50, wt:wt%). Electrospinning was carried out using a multinozzle electrospinning system with robot-controlled, movable and tunable spinnerets made by our laboratory (Fig.1a). Solutions A (at reduction time of 3 or 5.5 h) and B were each collected separately in 10 ml plastic syringe connected to two spinneret holders with 21G needle tip (inner diameter 510 nm). The two spinnerets were oriented horizontally on one side of the collector at an angle of 80 between them. The two syringes were xed in a syringe pump set at a constant feed rate of 1.0 mlh1. Each needle tip was connected to two separate power supplies with identical voltages of 15 kV. The grounded collector was made of aluminium plate covered with Teon sheet, and the tip-to-collector distance was kept at 20 cm for all tests. For comparison, neat PU nanober was also prepared by conventional single nozzle electrospinning, using the same conditions above. After

electrospinning, the nanobrous membrane was dried in a dry oven at 60  C for 48 h. The morphological structure of the samples was characterized by eld emission scanning electron microscopy (FESEM, Hitachi S-4800), and transmission electron microscopy (TEM, H7650, Hitachi). The samples for FESEM were coated with a very thin layer of platinum before characterization. Samples for TEM were prepared by electrospinning directly on a copper grid mesh with carbon coating for 20e30 s. Elemental analysis was carried out by energy dispersive X-ray spectroscopy (EDS). The ber diameter and nanoparticle size distribution were determined using ImageJ (NIH, USA) software, getting the average of 100 measurements based from the FESEM images. Fourier transform infrared (FTIR) spectra of the samples were obtained using a Paragon 1000 Spectrometer (Perkin Elmer) at a signal resolution of 1 cm1 within the range of 400e4000 cm1. The ultraviolet (UV)evisible spectra of the samples were obtained with a Lambda 900 UVevis spectrometer (PerkineElmer) over the range of 200e800 nm. The antibacterial activity of composite nanobers against Escherichia coli, a Gram-negative bacteria, was investigated by a zone inhibition method. The nanober mat samples were formed into circular discs with a diameter of 6 mm. Using a spread plate method, nutrient agar plates were inoculated with 1 ml of bacterial suspension containing around 105 colony forming units for each bacteria. The same amounts of samples were gently placed on the inoculated plates, and were then incubated at 37  C for 24 h. Zones of inhibition were determined by measuring the clear area formed around each sample. 3. Results and discussion Electrospinning of hybrid mats was carried out at two different reduction times of AgNO3 in PEO: 3 h and 5.5 h (referred herein as Hybrid 1 and Hybrid 2, respectively). A study by Saquing et al. [19] showed that high-molecular weight PEO has the capability of reducing AgNO3 into Ag NPs at ambient conditions. The reduction of AgNO3 to Ag NPs could be observed in the color change of the AgNO3/PEO/DW solution from milky white color at t 0 to dark green color at 3 h reduction time (Fig. 2d). The reduction of AgNO3 into Ag NPs in high MW PEO is attributed to the formation of pseudo-crown ethers that can bind silver ions within the coiled polyether helix, and these bound silver ions are reduced via oxidation of PEO fragments by the metal center [19,20]. The reason

Fig.1. (a) Schematic of the present angled two-spinneret electrospinning set-up; photographic images of (b) Hybrid 2 mat and; (c) neat PU nanobrous mat.

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Fig. 2. FESEM and TEM images of nanobrous mats of (a) neat PU; (b, e) Hybrid 1; (c, f) Hybrid 2, and; (d) photos of Ag/PEO solutions at reduction times of 0 and 3 h.

for the conformation similar to that of the crown ethers is the ionedipole interaction between the metal ion and the electron pair of the ether oxygen in the PEO fragments [21]. Furthermore, high MW PEO has longer polymer chains, which in turn can form more crown ether cavities and provide more reaction sites thus making it faster in reduction rate, comparing to using low MW PEO, which require at least 20 days of reduction time as reported elsewhere [20]. The electrospun nanobrous mats (Fig. 2aec) all showed highly-porous structure, and randomly-oriented, ultrane nanobers [22]. The neat PU nanobers showed continuous and smooth brous structure with an average ber diameter of 950 270 nm (Table 1). On the other hand, the hybrid Ag/PEO e PU nanobers showed two distinct ber sizes, i.e., larger ber sizes for PU nanobers and much smaller sizes for PEO nanobers. In
Table 1 Size distribution of electrospun neat PU and hybrid mats, and Ag NPs. Neat PU Fiber diameter (nm) Ag NP size (nm) PU PEO 950 270 e e Ag/PEO e PU (3 h) (Hybrid 1) 800 290 194 40 82 Ag/PEO e PU (5.5 h) (Hybrid 2) 770 280 200 36 90 30

Hybrid 1 (reduction time of 3 h, Fig. 2b), the PU and PEO nanobers showed an average diameter of 800 290 nm and 194 40 nm, respectively. A closer look at the PEO nanober (inset of Fig. 2b) shows very small, well-dispersed Ag NPs with diameters ranging from 6 to 10 nm. These Ag NPs were veried by TEM imaging as shown in Fig. 2e, showing highly-dispersed Ag NPs with narrow size distribution along the nanober. Similar structures in Hybrid 1 were observed in Hybrid 2 (Fig. 2c and f) but the Ag NPs were larger in diameter (d 90 30 nm) and had wider diameter distribution. These large Ag NP sizes are attributed to the aggregation of metallic silvers to form clusters, due to their high surface energy [23]. The increase in NP size is also supported by the UVevis spectroscopy results (Fig. 3b). Here, the duration of reduction time affected the sizes of the Ag NPs, wherein the longer reduction time produced bigger Ag NPs, which is consistent with the results of other studies [19]. A recent study reported that the reduction of AgNO3 into Ag NPs starts as early as 30 min and complete reduction is attained after 8 h, i.e., very little additional Ag NP formation is observed at reduction time from 8 h until 24 h. When the silver ions are reduced into metallic silver, the silver atoms aggregate into clusters which possess very high surface energy, thereby aggregating more into larger particles [23]. Thus, the longer reduction time, there is more chance of clusters to

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Fig. 3. (a) FTIR and (b) UVevis spectra of nanobrous mats of neat PU, Hybrid 1 and Hybrid 2; and (c) EDS spectra of Hybrid 2 mat.

aggregate more and form into bigger particles as observed in the present study. To analyze the composition of electrospun nanobers, we took FTIR spectra of the nanobrous mats (Fig. 3a). The characteristic peaks of neat PU nanobers can be assigned as: 2955 cm1 (CH2 asymmetric vibration); 2881 cm1 (CH3 symmetric vibrations); 1733 cm1 (free C]O); 1709 cm1 (C]O bond); 1532 cm1 (urethane amide II); 1081 cm1 C(O)eOeC stretching of the hard segment; and 821 cm1 (bending vibration in benzene ring) [24,25]. The typical absorption bands for PEO are 2894 cm1 (CH2 stretching), CeOeC stretching vibration at 1153, 1101, and 963 cm1, and at 1343 cm1 assigned as CeH deformation mode [26]. All peaks of neat PU were detected in the hybrid mats, and the additional peaks in Fig. 3a are attributed to the peaks of PEO. However, the band at 1478 (eCeH bending mode) of neat PU was observed to have shifted to lower frequency in the hybrid mats, indicating the formation of stronger hydrogen bond between PU and PEO [27]. These FTIR spectra results conrm the successful fabrication of bicomponent hybrid nanobrous mat consisting of PU and PEO nanober matrices. In order to verify the presence of Ag NPs on the hybrid nanobrous mats, EDS and UVevis spectra were obtained. In the present study, PEO was used as the in situ reductant of AgNO3 to form Ag NPs. A change of color was observed visually on the nanober mat from white (for neat PU, Fig. 1c) to yellowish mat (for the hybrid mats, Fig. 1b), which conrms the reduction of Ag ions and the formation of metallic silver nuclei [28]. The EDS spectra (inset of Fig. 3b) show Ag peak for the hybrid nanobrous mats, conrming

the presence of Ag NPs. The UVevis spectra (Fig. 3b) show peaks from 400 to 470 nm, which signies the surface plasmon band resonance of Ag NPs [19,29], while no peak absorbance was obtained for neat PU in this region. Hybrid 1 showed a peak at 415 nm, while Hybrid 2 shifted its peak to a longer wavelength (i.e., 425 nm) (blue shift), signifying an increase in NP sizes [30], which corresponds well with the sizes observed in SEM and TEM images (Fig. 2), and as also observed by other studies [31]. Fig. 4 shows the antibacterial activity of the electrospun nanobrous mats via a zone inhibition test. Previous studies observed that the antimicrobial activity of Ag NPs depended on their average size [32]. It was reported that Ag NPs with sizes of 1e10 nm can disturb the permeability of bacteria and respiratory functions, thus can efciently inactivate them [33,34]. In this study, Hybrid 1 nanobrous mat, which contains Ag NPs with average diameter of 8 nm (Fig. 4c) showed much higher bacterial inactivation as compared to Hybrid 2 (Ag NP average diameter 90 nm) (Fig. 4d), while individual nanobrous mats of neat PU (Fig. 4a) and neat PEO (Fig. 4b) alone did not show any signs of antibacterial activity. This suggests that without the aid of Ag NPs, the individual neat nanobrous components of PU and PEO could not provide any antibacterial effect. The higher bacterial activity of Hybrid 1 could be attributed to the presence of smaller Ag NPs, which provide large surface area to come in contact with bacteria, when compared with bigger Ag NPs in Hybrid 2 [35]. Several other researchers reported high bacterial inactivation efciency of Ag NP/polymer composites with Ag NP sizes ranging from 2 to 21 nm [23,32,36,37].

Fig. 4. Zone of inhibition test against E. coli bacteria for nanobrous mats of (a) neat PU, (b) neat PEO, (c) Hybrid 1, and (d) Hybrid 2 (scale bar 5 mm).

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4. Conclusion In summary, we have demonstrated a facile way to fabricate hybrid nanobrous mats with antibacterial functionality by dualspinneret electrospinning. Using PEO both as reducing agent of AgNO3 and template for the formed Ag NPs, we can utilize the dualspinneret electrospinning to selectively functionalize the component nanobers for making new materials. Here, the larger PU nanobers give structural stability and prolonged use of the hybrid materials, while the thinner PEO nanobers provide more reactive surface area, and the decoration of Ag NPs on PEO surfaces without the use of chemicals gives antibacterial functionality to the hybrid mat. The reduction time of AgNO3 in PEO plays an important role in the size control of the Ag NPs. The hybrid mat with Ag NPs of 8 nm average size (i.e., reduction time of 3 h) exhibited strong antibacterial activity. The present hybrid mat has great potential for antibacterial tissue scaffolds or for wound-healing applications. Acknowledgement This research was supported by a Grant from the Ministry of Education, Science and Technology of Korea through the National Research Foundation (Project no. 2011-0011807). References
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