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SilVer Nanoparticle Based Formulation for Topical Use articles Figure 1. UV-vis absorption spectra of SNP suspension. Figure 2. HRTEM of individual SNP. Table 1. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of SNP against Different Organisms Figure 3. Kill curve of reference bacteria and MDR cultures at MIC of SNP (error bars not shown as deviations from the mean values are <0.1 × log 10). Table 2. Microbicidal Activity of SNP at MIC and MBC against Different Bacterial Strains SNP (μg/mL) MIC50a MIC90b MBC99.9c organism Pseudomonas aeruginosa ATCC 9027 Escherichia coli ATCC 117 Klebsiella aerogenes ATCC 1950 Salmonella abony NCTC 6017 Salmonella typhimurium ATCC 23564 Proteus vulgaris NCI B 4157 Bacillus subtilis ATCC 6633 Staphylococcus epidermidis ATCC 12228 Staphylococcus aureus ATCC 6538 Pseudomans sp. # MDR 1 Staphylococcus sp. # MDR 1 D at MICa (h) D* at 3.5 4.4 6.7 6.4 5.4 10 7.17 7.3 18.9 5.6 >24 MBCb (h) 4.7 6 9 9 7 8 12 12 >24 4.5 >24 organisms used Pseudomonas aeruginosa ATCC 9027 3.12 6.25 12.5 Salmonella abony NCTC 6017 3.12 6.25 12.5 Salmonella typhimurium ATCC 23564 3.12 6.25 12.5 Klebsiella aerogenes ATCC 1950 1.56 6.25 12.5 Proteus vulgaris NCIB 4157 3.12 6.25 12.5 Bacillus subtilis ATCC 6633 3.12 6.25 12.5 Staphylococcus aureus ATCC 6538 6.25 12.5 ND Staphylococcus epidermidis ATCC 12228 3.12 6.25 12.5 Escherichia coli ATCC 117 1.56 6.25 12.5 a D ) time required to achieve 1 log10 reduction total viable cell count. b D* ) time required to achieve 3 log10 reduction total viable cell count. As further extension of the MIC and MBC, time kill studies were initiated. Figure 3 demonstrates time-dependent microbicidal action of SNP at MIC. It is evident that Gram- negative bacteria are killed faster than Gram-positive bacteria and there is a linear relationship between log CFU versus time. Slope of the line of regression (K) could be used for the analysis of bactericidal activity of SNP against different microbial species. Data in Table 2 show that the time required to achieve a 1 log10 reduction (D) in viable cells differs with the organisms indicating their varying sensitivity to SNP. At MIC (6.25 μg/mL) time required to achieve 1 log10 reduction (90%) in all tested strains (except S. aureus) ranged between 3.5 and 10 h. In general, Gram-negative bacteria were at the lower end and Gram-positive bacteria were at a MIC50: minimum inhibitory concentration required to kill 50% of population (scored visually after 14-16 h of incubation). b MIC90: minimum inhibitory concentration required to kill 90% of population (scored visually after 24 h of incubation). c MBC: minimum bactericidal concentration required to kill 99.9% of population (scored by spot inoculation on MH agar from the wells showing no growth after 24 h). ND: not detectable. determined because SNP was found to be bacteriostatic even at the highest concentration available for testing, i.e., 50 μg/ mL. An analysis of MIC values for standard cultures and MDR strains indicated that the majority of strains, i.e., 66% showed MICs e 3.12 μg/mL. Out of these, 4% strains display MIC as low as 0.78 μg/mL. These results show that SNP have a potent antimicrobial activity at very low concentrations. VOL. 6, NO. 5 MOLECULAR PHARMACEUTICS 1393PDF Image | Silver Nanoparticles in Therapeutics: Antimicrobial Gel
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