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SilVer Nanoparticle Based Formulation for Topical Use articles Figure 11. Confocal micrographs of acridine orange/ethidium bromide stained Hep G2 cells. (A) Unexposed (control) cells have a normal green nucleus indicating live cells. (B) Cells exposed to ∼(1/2)IC50 SNP (125 μg/mL) for 24 h display bright green nucleus with condensed or fragmented chromatin suggesting apoptosis. (C) Cells exposed to ∼2 × IC50 SNP (500 μg/mL) for 24 h have a structurally normal orange nucleus indicative of necrosis. Scale bar indicates 100 μm. Table 5. Quantification of AO/EB Stained Hep G2 Cells (%)a after Exposure to SNP (∼(1/2)IC50 SNP and 2 × IC50) for 24 h A study by Panacek et al.19 described synthesis of silver nanoparticles of varying sizes (but narrow size distributions) using four different saccharides. The antimicrobial property of these nanoparticles against many Gram-positive and Gram- negative bacteria including multidrug-resistant strains was found to be size dependent. Nanoparticles with size >25 nm exhibited MIC of 6.75-54 μg/mL whereas 25 nm size particles showed MIC in the range of 1.69-13.5 μg/mL. The MIC results obtained with 25 nm particles are comparable to those obtained by us in the present study, where SNP suspension containing 7-20 nm particles were used. This is an important result, particularly when antibiotic resistance among bacterial species is increasing at an alarming rate and very few alternative options are available to address the issue. Park et al.20 showed that hybrid silver (Ag) nanoparticles (size 3-7 nm) loaded on SiO2 nanoparticles inhibited a range of standard fungi at concentration of 1 ppm. However, in our study, the obtained antifungal concentrations are 25-fold (∼25 μg/mL). An interesting observation on variation in antimicrobial activity of silver nanoparticles due to strain specificity has been discussed by Ruparelia et al.21 They showed that among the tested strains of Escherichia coli (four strains), Bacillus subtilis and Staphylococcus aureus (three strains), Escherichia coli strains showed maximum variation in the silver nanoparticle induced antimicrobial activity. Our results corroborate these findings as we find a lot of strain- dependent variation in antimicrobial response of SNP activ- ity. In another study the MIC and MBC of Acticoat with S. aureus, S. epidermidis, E. coli, K. aerogenes and P. aerugi- nosa was reported to be 5-12.5 μg/mL,22,23 which compares well with the results obtained with SNP. It was reported that (19) Panacek, A.; Kvitek, L.; Prucek, R.; Kolar, M.; Vecerova, R.; Pizurova, N.; Sharma, V. K.; Nevecna, T.; Zboril, R. Silver colloid nanoparticles: synthesis, characterization, and their antibacterial activity. J. Phys. Chem. B 2006, 110, 16248–16253. (20) Park, H.-J.; Kim, H. J.; Kim, S. H.; Oh, S.-D.; Choi, S.-H. Radiolytic synthesis of hybrid silver nanoparticles and their biobehavior. Key Eng. Mater. 2007, 342-343, 897–900. (21) Ruparelia, J. P.; Chatterjee, A. K.; Duttagupta, S. P.; Mukherji, S. Strain specificity in antimicrobial activity of silver and copper nanoparticles. Acta Biomaterialia 2008, 4, 707–716. (22) Burrel, R. E.; McIntosh, C. L.; Morris, I. R. Process of activating antimicrobials materials. 1995, US Patent 5,455,886. type of cells live cells apoptotic cells necrotic cells SNP (125 μg/mL) SNP (500 μg/mL) control (unexposed) cells 96 ( 3 3 ( 2 1 ( 1 exposed to ∼(1/2)IC50 exposed to ∼2 × IC50 a The data are expressed as mean ( standard deviation (SD) of three independent experiments. Asterisk (*) denotes a statistically significant difference compared to control (p < 0.05). nanoparticles. Cho et al.16 reported that poly-(N-vinyl-2- pyrrolidone) (PVP) stabilized silver nanoparticles showed MIC of 5 μg/mL against S. aureus and 10 μg/mL against E. coli. S. aureus and E. coli were completely inhibited at 50 and 100 μg/mL silver. Raffi et al.17 reported that silver nanoparticles (mean size 16 nm) synthesized by inert gas condensation (IGC) method were effective bactericide against Escherichia coli, at concentration 60 μg/mL and higher. In another study, Shahverdi et al.18 synthesized silver nano- particles using culture supernatants of Klebsiella pneumoniae for reduction of aqueous Ag+ ions. The antimicrobial activity of silver nanoparticles against Staphylococcus aureus and Escherichia coli was assessed by diffusion assay. It was also observed that activities of various antibiotics (penicillin G, amoxicillin, erythromycin, clindamycin, and vancomycin) were increased in the presence of silver nanoparticles. In the present study, MIC results obtained with SNP are comparable to those reported by Cho et al.16 for PVP stabilized silver nanoparticles. (16) Cho, K. H.; Park, J. E.; Osaka, T.; Park, S. G. The study of antimicrobial activity and preservative effects of nanosilver ingredient. Electrochim. Acta 2005, 51, 956–60. (17) Raffi, M.; Hussain, F.; Bhatti, T. M.; Akhter, J. I.; Hameed, A.; Hasan, M. M. Antibacterial Characterization of Silver Nanopar- ticles against E. coli ATCC-15224. J. Mater. Sci. Technol. 2008, 24, 192–196. (18) Shahverdi, A. R.; Fakhimi, A.; Shahverdi, H. R.; Minaian, S. Synthesis and effect of silver nanoparticles on the antibacterial activity of different antibiotics against Staphylococcus aureus and Escherichia coli. Nanomed.: Nanotechnol., Biol., Med. 2007, 3, 168–171. 67 ( 5* 28 ( 5* 5 ( 1* 53 ( 6* 11 ( 3* 36 ( 3* VOL. 6, NO. 5 MOLECULAR PHARMACEUTICS 1397PDF Image | Silver Nanoparticles in Therapeutics: Antimicrobial Gel
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