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Summary: A feasible method for the preparation of antimicrobial ultrane bers with silver nanoparticles was developed by direct electrospinning of a cellulose acetate (CA) solution with small amounts of silver nitrate followed by photoreduction. Silver nanoparticles in ultrane CA bers were stabilized by interactions with carbonyl oxygen atoms in CA. Ultrane CA bers with silver nanoparticles showed very strong antimicrobial activity.

TEM image of an ultrane CA ber electrospun from 10 wt.-% CA solution with 0.5 wt.-% AgNO3.

Preparation of Antimicrobial Ultrane Cellulose Acetate Fibers with Silver Nanoparticles

Won Keun Son,1 Ji Ho Youk,2 Taek Seung Lee,3 Won Ho Park*3
1 2

Research Institute of Advanced Materials, Chungnam National University, Daejeon 305-764, South Korea Department of Textile Engineering, Inha University, Incheon 402-751, South Korea 3 Department of Textile Engineering, Chungnam National University, Daejeon 305-764, South Korea Fax: (82) 42-823-3736; E-mail: parkwh@cnu.ac.kr

Received: July 20, 2004; Revised: August 9, 2004; Accepted: August 9, 2004; DOI: 10.1002/marc.200400323 Keywords: antimicrobial activity; biological applications of polymers; cellulose acetate; bers; nanoparticles

Introduction
Silver ions and silver compounds have been widely used in various biomedical elds, such as wound dressing materials, body wall repairs, augmentation devices, tissue scaffolds, and antimicrobial lters.[17] Positively charged silver ions attracted to electronegative bacterial cells are bonded to their membrane or bacterial DNAwith sulfhydryl groups, resulting in the prevention of proliferation of bacteria and blockage of biolm production. Microorganisms with resistance to the antimicrobial activity of silver are exceedingly rare. In recent years, ultrane polymer bers prepared by electrospinning of a polymer solution have been drawing great attention because of their unique properties, such as high surface area-to-volume and length-to-diameter ratios. When a strong electrostatic force is applied to the capillary
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containing a polymer solution, it is ejected from the capillary and deposited as a nonwoven brous mat on a template serving as the ground for the electric charges. It is expected that antimicrobial ultrane bers will exhibit much stronger antimicrobial activity than conventional microbers because of their high surface area-to-volume ratio. Yang et al. rst prepared ultrane polyacrylonitrile (PAN) bers containing silver nanoparticles by electrospinning.[8] The average diameters of the PAN bers and silver nanoparticles were 400 and 100 nm, respectively. It was recently reported that silver nanoparticles also had very strong antimicrobial activity.[7,9,10] In this study, antimicrobial ultrane cellulose acetate (CA) bers with silver nanoparticles were prepared by electrospinning of a CA solution with small amounts of silver nitrate (AgNO3) followed by slow and fast photoreduction.
2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

DOI: 10.1002/marc.200400323

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Experimental Part
Ultrane CA (acetyl content 39.8%, M n 30 000) bers were electrospun from a CA solution in a mixed solvent of acetone/ water (80:20, w/w). In order to prepare ultrane CA bers containing silver ions, 10 wt.-% CA solutions with small amounts of AgNO3 (0.010.5 wt.-% to the amounts of CA) in acetone/water (80:20, w/w) were electrospun. The electrospinning setup consisted of a syringe and needle (ID 0.84 mm), a ground electrode (d 21.5 cm, a stainless steel sheet on a drum whose rotation speed can be varied), and a high voltage supply (Chungpa EMT, CPS-40K03), as used in our previous studies.[1113] CA solutions were electrospun at 17 kV positive voltage, 10 cm working distance (the distance between the needle tip and the ground electrode), and 3 mL h1 solution ow rate. The morphologies of the ultrane CA bers were observed on a scanning electron microscope (SEM) (Hitachi S-2350) after gold coating. The average diameters were determined by analyzing SEM images with a custom code image analysis program. Transmission electron microscopy (TEM) images were obtained by using a JEOL JEM-2010 for samples deposited on carbon coated copper grids. The viscosities of CA solutions were measured by a Brookeld digital viscometer (Model DV-E) at 25 8C. The conductivities and

surface tensions were measured by a conductivity meter (Isteck Model 455C) and by an interfacial tension meter (Kruss Model K8) at 25 8C, respectively. A 500 W high-pressure mercury lamp (l 310380 nm) was used for the UVirradiation source. X-ray photoelectron spectroscopy (XPS) measurements were performed on a VG-Scientic ESCALAB 250 spectrometer with a monochromatic AlKa X-ray source at 1486.6 eV. Highresolution XPS spectra were recorded in 0.05 eV steps with a pass energy of 20 eV. The binding energy peaks were deconvoluted using Gaussian functions and a least squares curvetting program. Silver ions in ultrane CA bers were extracted with water and their concentrations were monitored by a Direct Reading Echelle inductively coupled plasma (ICP) spectrometer (Leeman Labs Inc., DRE). The antimicrobial activity of ultrane CA bers with and without silver nanoparticles was tested against Gram-positive Staphylococcus aureus (S. aureus, ATCC6538) and Gram-negative Escherichia coli (E. coli, ATCC 25922), Klebsiella pneumoniae (K. pneumoniae, ATCC 4352), and Pseudomonas aeruginosa (P. aeruginosa, ATCC 27853) by the nonwoven fabric attachment method.[14] The agar plates containing test samples and the control (blank) were incubated at 37 8C for 18 h. The reductions of bacteria were calculated according to the following equation, Reduction (%) (B A)/B 100, where A and B

Figure 1. SEM images of ultrane CA bers electrospun from 10 wt.-% CA solutions with different amounts of AgNO3: (a) 0 wt.-%, (b) 0.05 wt.-%, (c) 0.3 wt.-%, and (d) 0.5 wt.-%.
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W. K. Son, J. H. Youk, T. S. Lee, W. H. Park

are the surviving cells (colony forming unit mL1) for the plates containing test samples and the control, respectively, after 1 h contact time.

Results and Discussion

In our previous study, a mixed solvent system of acetone/ water was developed for the continuous electrospinning of a CA solution.[11,12] When acetone was used alone, the tip of the needle was easily blocked by its rapid evaporation. The water contents for the continuous electrospinning of 17 wt.-% CAwere in the range of 1015 wt.-%. However, in this study, 20 wt.-% of water was used to completely dissolve AgNO3 and the concentration of CA was lowered instead. In order to determine the minimum electrospinning concentration of CA in acetone/water (80:20, w/w), solution viscosities of CA were measured according to the CA concentration. The solution viscosities were rapidly increased from 10 wt.-% of CA because of the onset of chain entanglement. Practically, long uniform bers were electrospun at CA concentrations above 10 wt.-%. Figure 1 shows SEM images of ultrane CA bers electrospun from 10 wt.-% CA solutions with different amounts of AgNO3.

The average diameters of the CA bers electrospun with 0, 0.05, 0.3, and 0.5 wt.-% AgNO3 were 1 910, 680, 640, and 610 nm, respectively. The ber diameters were dramatically decreased when small amounts of AgNO3 were added. Figure 2 shows changes in solution properties of CA according to the amounts of AgNO3 added. The conductivities of CA solutions were linearly increased, while their viscosities and surface tensions were not changed with increasing the amounts of AgNO3. The addition of AgNO3 increased the charge density in CA solutions and thus stronger elongation forces were imposed on the ejected jets under the electrical eld, resulting in substantially straighter shaped and ner CA bers.[13,15] It is known that silver nanoparticles can be prepared by a UV irradiation photoreduction technique.[1618] The color of ultrane CA bers electrospun with AgNO3 was gradually changed to light yellow, indicating that silver ions in ultrane CA bers were slowly photoreduced and aggregated into silver nanoparticles in a general laboratory environment. Silver nanoparticles formed preferentially on the surface of the ultrane CA bers and had an almost spherical shape (Figure 3). The average diameters of the ultrane CA bers electrospun with 0.05 and 0.5 wt.-%

Figure 2. Changes in (a) viscosities, (b) surface tensions, and (c) conductivities of 10 wt.-% CA solutions according to the amounts of AgNO3 added.
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Figure 3. TEM images of ultrane CA bers electrospun from 10 wt.-% CA solutions with: (a) 0.05 wt.-%, (b) 0.5 wt.-% AgNO3, (c) 0.5 wt.-% AgNO3, and subsequently UV irradiated for 3 h. TEM images of (a) and (b) were taken 20 d after electrospinning.

AgNO3 were 3.3 and 6.9 nm, respectively. They were dependent on the amounts of AgNO3 added.[19] Figure 3c shows TEM images of ultrane CA bers electrospun with 0.5 wt.-% AgNO3 and subsequently UV irradiated for 3 h. A large number of silver nanoparticles with an average diameter of 15.4 nm were generated by the rapid photoreduction. Comparing Figure 3b and 3c, it is considered that only part of silver ions was reduced to silver nanoparticles in the laboratory environment. The nature of the interaction between CA and silver nanoparticles was investigated by XPS. Figure 4 shows
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XPS spectra of the C1s, O1s, and Ag regions for ultrane CA bers with silver nanoparticles. The C1s photoemissions of CA were not changed by the addition of AgNO3, implying that carbon atoms do not directly interact with the silver nanoparticles. The O1s photoemission spectra were shifted to a higher energy and resolved into two peaks. The lower peak is similar to that of pure CA and the higher one at 531.7 eV is attributed to the interaction between carbonyl oxygen atoms and silver nanoparticles. The shift to higher energy indicates that the carbonyl oxygen atoms have a lower electronic density than those in pure CA.[20,21] In
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W. K. Son, J. H. Youk, T. S. Lee, W. H. Park

Figure 4. C1s, O1s, and Ag3d XPS spectra for ultrane CA bers with different amounts of AgNO3: (a) C1s, (b) O1s, and (c) Ag3d. The XPS spectra were taken 20 d after electrospinning.

addition, Ag 3d5/2 and 3d3/2 binding energies in AgNO3 were shifted to lower energies of 367.0 and 373.0 eV, respectively, because of the photoreduction of silver ions.[2224] The antimicrobial activity of ultrane CA bers with silver nanoparticles was tested against S. aureus, E. coli, K. pneumoniae, and P. aeruginosa according to the nonwoven fabric attachment method. Bacterial colonies were not observed for ultrane CA bers with silver nanoparticles (electrospun with 0.5 wt.-% AgNO3 and slowly photoreduced), indicating silver nanoparticles and silver ions not reduced inhibited the growth of the bacteria. The concentration of these silver ions was analyzed by ICP spectroscopy. The concentration rapidly increased to 3% of the initial silver ions incorporated, and then slowly leveled off to 7% for 160 h. The reductions of bacteria for ultrane CA bers electrospun with 0.01 wt.-% of AgNO3, were 21.1, 23.2, 19.5, and 26.9% for S. aureus, K. pneumoniae, E. coli, and P. aeruginosa, respectively. However, for ultrane CA
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bers electrospun with 0.05 wt.-% of AgNO3, all bacteria were reduced to 99.9% after incubation. It was revealed that very small amounts of AgNO3 were needed to endow ultrane CA bers with very strong antimicrobial activity.

Conclusion
The average diameters of ultrane CA bers electrospun from a 10 wt.-% CA solution in acetone/water (80:20, w/w) with 0, 0.05, 0.3, and 0.5 wt.-% AgNO3 were 1 910, 680, 640, and 610 nm, respectively. Silver ions in ultrane CA bers were photoreduced into silver nanoparticles even in a general laboratory environment, which were stabilized by interactions with carbonyl oxygen atoms in the CA. The average diameters of the silver nanoparticles were in the range of 316 nm. The ultrane CA bers showed very strong antimicrobial activity attributable to the silver nanoparticles and nonreduced silver ions.
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Acknowledgements: This research was supported by the Korea Research Foundation (Grant KRF-2001-005-E00037) and by the Program for the Training of Graduate Students in Regional Innovation, which was conducted by the Ministry of Industry and Energy of the Korean Government.

[1] B. Jansen, M. Rinck, P. Wolbring, A. Strohmeier, T. Jahnns, J. Biomater. Appl. 1994, 9, 55. [2] H. Y. Kang, M. J. Jung, Y. K. Jeong, Korean J. Biotechnol. Bioeng. 2000, 15, 521. [3] Q. L. Feng, J. Wu, G. Q. Chen, F. Z. Cui, T. N. Kim, J. O. Kim, J. Biomed. Mater. Res. 2000, 52, 662. [4] U. Klueh, V. Wagner, S. Kelly, A. Johnson, J. D. Bryers, J. Biomed. Mater. Res. 2000, 53, 621. [5] D. C. Clupper, L. L. Hench, J. Mater. Sci. Mater. Med. 2001, 12, 917. [6] J. P. Guggenbichler, Mat.-wiss. u. Werkstofftech. 2003, 34, 1145. [7] I. Sondi, B. Salopek-Sondi, J. Colloid Interface Sci. 2004, 275, 177. [8] Q. B. Yang, D. M. Li, Y. L. Hong, Z. Y. Li, C. Wang, S. L. Qiu, Y. Wei, Synth. Met. 2003, 137, 973. [9] L. Balogh, D. R. Swanson, D. A. Tomalia, G. L. Hagnauer, A. T. McManus, Nano Lett. 2001, 1, 18.

[10] J. Dai, M. L. Bruening, Nano Lett. 2002, 2, 497. [11] W. K. Son, J. H. Youk, T. S. Lee, W. H. Park, J. Polym. Sci., Part B: Polym. Phys. 2004, 42, 5. [12] W. K. Son, J. H. Youk, W. H. Park, Biomacromolecules 2004, 5, 197. [13] W. K. Son, J. H. Youk, T. S. Lee, W. H. Park, Polymer 2004, 45, 2959. [14] Antibacterial of Polymeric Materials (FC-TM-20), FITI Testing & Research Institute, 2001. [15] Z. Xinhua, K. Kim, R. Shaofeng, B. S. Hsiao, B. Chu, Polymer 2002, 43, 4403. [16] W. Wang, S. A. Asher, J. Am. Chem. Soc. 2001, 123, 12528. [17] Z. Zhang, L. Zhang, S. Wang, W. Chen, Y. Lei, Polymer 2001, 42, 8315. [18] H. W. Lu, S. H. Liu, X. L. Wang, X. F. Qian, J. Yin, Z. K. Zhu, Mater. Chem. Phys. 2003, 81, 104. [19] Z. Zhang, M. Han, J. Mater. Chem. 2003, 13, 641. [20] H. H. Huang, X. P. Ni, G. L. Loy, C. H. Chew, K. L. Tan, F. C. Loh, J. F. Deng, G. Q. Xu, Langmuir 1996, 12, 909. [21] U. P. Rodrigues-Filho, Y. Gushikem, M. C. Goncalves, R. C. Cachichi, Chem. Mater. 1996, 8, 1375. [22] M. Pattabi, K. M. Rao, S. R. Sainkar, M. Sastry, Thin Solid Films 1999, 338, 40. [23] S. He, J. Yao, S. Xie, S. Pang, H. Gao, Chem. Phys. Lett. 2001, 343, 28. [24] Y. Tan, Y. Wang, L. Jiang, D. Zhu, J. Colloid Interface Sci. 2002, 249, 336.

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