Comparison of methods to detect the in vitro activity of silver nanoparticles (AgNP) against multidrug resistant bacteria

Journal of Nanobiotechnology

Silvia Figueiredo Costa⁵  Anna Sara Shafferman Levin³  Valtencir Zucolotto⁴  Valeria Spolon Marangoni⁴  Fabiane Fantinelli Franco⁴  Roberto Angelo de Oliveira¹  Marcelo Martins Seckler¹  José Pinhata Otoch²  Luiz Francisco Poli de Figueiredo²  Emerson Danguy Cavassin³ 

[1] Faculty of Engenier São Paulo, University of São Paulo, São Paulo, Brazil;Department of Surgery São Paulo, University of São Paulo, São Paulo, Brazil;Department of Infectious Diseases, University of São Paulo, São Paulo, Brazil;Nanomedicine and Nanotoxicology Group, University of São Paulo, IFSC, São Paulo, Brazil;LIM-54 (Laboratório de Bacteriologia), Instituto de Medicina Tropical, Av. Dr. Enéas de Carvalho Aguiar, 470, Prédio II, 1º andar, Sala 112, Cerqueira César, São Paulo 054503-00, SP, Brazil

关键词: Multidrug resistant bacteria;    Antimicrobial test;    Silver nanoparticle;   

Others  :  1230500
DOI  :  10.1186/s12951-015-0120-6

 received in 2014-09-29, accepted in 2015-09-09,  发布年份 2015

【 摘 要 】

Background

Multidrug resistant microorganisms are a growing challenge and new substances that can be useful to treat infections due to these microorganisms are needed. Silver nanoparticle may be a future optionfor treatment of these infections, however, the methods described in vitro to evaluate the inhibitory effect are controversial.

Results

This study evaluated the in vitro activity of silver nanoparticles against 36 susceptible and 54 multidrug resistant Gram-positive and Gram-negative bacteria from clinical sources. The multidrug resistant bacteria were oxacilin-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus spp., carbapenem- and polymyxin B-resistant A. baumannii, carbapenem-resistant P. aeruginosa and carbapenem-resistant Enterobacteriaceae. We analyzed silver nanoparticles stabilized with citrate, chitosan and polyvinyl alcohol and commercial silver nanoparticle. Silver sulfadiazine and silver nitrate were used as control. Different methods were used: agar diffusion, minimum inhibitory concentration, minimum bactericidal concentration and time-kill. The activity of AgNPs using diffusion in solid media and the MIC methods showed similar effect against MDR and antimicrobial-susceptible isolates, with a higher effect against Gram-negative isolates. The better results were achieved with citrate and chitosan silver nanoparticle, both with MIC ₉₀of 6.75 μg mL ⁻¹ , which can be due the lower stability of these particles and, consequently, release of Ag ⁺ions as revealed by X-ray diffraction (XRD). The bactericidal effect was higher against antimicrobial-susceptible bacteria.

Conclusion

It seems that agar diffusion method can be used as screening test, minimum inhibitory concentration/minimum bactericidal concentration and time kill showed to be useful methods. The activity of commercial silver nanoparticle and silver controls did not exceed the activity of the citrate and chitosan silver nanoparticles. The in vitro inhibitory effect was stronger against Gram-negative than Gram-positive, and similar against multidrug resistant and susceptible bacteria, with best result achieved using citrate and chitosan silver nanoparticles. The bactericidal effect of silver nanoparticle may, in the future, be translated into important therapeutic and clinical options, especially considering the shortage of new antimicrobials against the emerging antimicrobial resistant microorganisms, in particular against Gram-negative bacteria.

【 授权许可】

   
2015 Cavassin et al.

【 图 表 】

Fig.1.

Fig.2.

Fig.3.

Fig.4.

Fig.5.

Fig.6.

Fig.7.

Fig.8.

Fig.9.

【 参考文献 】

• [1]Planquette B, Timsit JF, Misset BY, Schwebel C, Azoulay E, Adrie C, Vesin A, Jamali S, Zahar JR, Allaouchiche B et al.. Pseudomonas aeruginosa ventilator-associated pneumonia predictive factors of treatment failure. Am J Respir Crit Care Med. 2013; 188:69-76.
• [2]Webb GF, D’Agata EMC, Magal P, Ruan S. A model of anti biotic-resistant bacterial epidemics in hospitals. Proc Natl Acad Sci USA. 2005; 102:13343-13348.
• [3]Lara HH, Ayala-Nunez NV, Turrent LDI, Padilla CR. Bactericidal effect of silver nanoparticles against multidrug-resistant bacteria. World J Microbiol Biotechnol. 2010; 26:615-621.
• [4]Morones JR, Elechiguerra JL, Camacho A, Holt K, Kouri JB, Ramirez JT, Yacaman MJ. The bactericidal effect of silver nanoparticles. Nanotechnology. 2005; 16:2346-2353.
• [5]Ansari MA, Khan HM, Khan AA, Malik A, Sultan A, Shahid M, Shujatullah F, Azam A. Evaluation of antibacterial activity of silver nanoparticles against MSSA and MRSA on isolates from skin infections. Biol Med. 2011; 3:141-146.
• [6]Gong P, Li H, He X, Wang K, Hu J, Tan W, Tan S, Zhang XY. Preparation and antibacterial activity of Fe 3 O 4 @Ag nanoparticles. Nanotechnology. 2007; 18:604-611.
• [7]Chaloupka K, Malam Y, Seifalian AM. Nanosilver as a new generation of nanoproduct in biomedical applications. Trends Biotechnol. 2010; 28:580-588.
• [8]ISO/TS16550:2014: Nanotechnologies—determination of silver nanoparticles potency by release of muramic acid from Staphylococcus aureus.
• [9]ISO/TR16197:2014: Nanotechnologies—compilation and description of toxicological screening methods for manufactured nanomaterials.
• [10]Mirzajani F, Ghassempour A, Aliahmadi A, Esmaeili MA. Antibacterial effect of silver nanoparticles on Staphylococcus aureus. Res Microbiol. 2011; 162:542-549.
• [11]Singh K, Panghal M, Kadyan S, Chaudhary U, Yadav JP. Green silver nanoparticles of Phyllanthus amarus: as an antibacterial agent against multi drug resistant clinical isolates of Pseudomonas aeruginosa. J Nanobiotechnol. 2014;12:40.
• [12]Kalishwaralal K, BarathManiKanth S, Pandian SRK, Deepak V, Gurunathan S. Silver nanoparticles impede the biofilm formation by Pseudomonas aeruginosa and Staphylococcus epidermidis. Colloids Surf B-Biointerf. 2010; 79:340-344.
• [13]Pirnay JP, De Vos D, Cochez C, Bilocq F, Pirson J, Struelens M, Duinslaeger L, Cornelis P, Zizi M, Vanderkelen A. Molecular epidemiology of Pseudomonas aeruginosa colonization in a burn unit: persistence of a multidrug-resistant clone and a silver sulfadiazine-resistant clone. J Clin Microbiol. 2003; 41:1192-1202.
• [14]Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically; Approved Standard—Ninth Edition. CLSI document M07-A9 (ISBN 1-56238-783-9 [Print]; ISBN 1-56238-784-7 [Electronic]). In Clinical and Laboratory Standards Institute. 950 West Valley Road, Suite 2500, Wayne, Pennsylvania 19087, USA: Clinical and Laboratory Standards Institute; 2012.
• [15]Pileni MP. Nanosized particles made in colloidal assemblies. Langmuir. 1997; 13:3266-3276.
• [16]Solomon SD, Bahadory M, Jeyarajasingam AV, Rutkowsky SA, Boritz C, Mulfinger L. Synthesis and study of silver nanoparticles. J Chem Educ. 2007; 84:322-325.
• [17]Sharma VK, Yngard RA, Lin Y. Silver nanoparticles: green synthesis and their antimicrobial activities. Adv Colloid Interface Sci. 2009; 145:83-96.
• [18]Coronado EA, Encina ER, Stefani FD. Optical properties of metallic nanoparticles: manipulating light, heat and forces at the nanoscale. Nanoscale. 2011; 3:4042-4059.
• [19]Ruparelia JP, Chatteriee AK, Duttagupta SP, Mukherji S. Strain specificity in antimicrobial activity of silver and copper nanoparticles. Acta Biomater. 2008; 4:707-716.
• [20]Zhang JT, Li XL, Sun XM, Li YD. Surface enhanced Raman scattering effects of silver colloids with different shapes. J Phys Chem B. 2005; 109:12544-12548.
• [21]Shameli K, Bin Ahmad M, Yunus W, Rustaiyan A, Ibrahim NA, Zargar M, Abdollahi Y. Green synthesis of silver/montmorillonite/chitosan bionanocomposites using the UV irradiation method and evaluation of antibacterial activity. Int J Nanomed. 2010; 5:875-887.
• [22]Philip D. Biosynthesis of Au, Ag and Au-Ag nanoparticles using edible mushroom extract. Spectrochimica Acta Part a Mol Biomol Spectrosc. 2009; 73:374-381.
• [23]Shankar SS, Ahmad A, Sastry M. Geranium leaf assisted biosynthesis of silver nanoparticles. Biotechnol Prog. 2003; 19:1627-1631.
• [24]Wang HS, Qiao XL, Chen JG, Ding SY. Preparation of silver nanoparticles by chemical reduction method. Colloids Surf a-Physicochem Eng Asp. 2005; 256:111-115.
• [25]Stebounova LV, Adamcakova-Dodd A, Kim JS, Park H, O’Shaughnessy PT, Grassian VH, Thorne PS. Nanosilver induces minimal lung toxicity or inflammation in a subacute murine inhalation model. Particle Fibre Toxicol 2011; 8.
• [26]Ayala-Núñez NV, Villegas HL, Turrent LC, Padilla CR. Silver nanoparticles toxicity and bactericidal effect against methicillin-resistant Staphylococcus aureus : nanoscale Does Matter. NanoBiotechnology. 2009; 5:2-9.
• [27]Jena P, Mohanty S, Mallick R, Jacob B, Sonawane A. Toxicity and antibacterial assessment of chitosan-coated silver nanoparticles on human pathogens and macrophage cells. Int J Nanomed. 2012; 7:1805-1818.
• [28]Shrivastava S, Bera T, Roy A, Singh G, Ramachandrarao P, Dash D. Characterization of enhanced antibacterial effects of novel silver nanoparticles. Nanotechnology 2007; 18.
• [29]Patil RS, Kokate MR, Jambhale CL, Pawar SM, Han SH, Kolekar SS. One-pot synthesis of PVA-capped silver nanoparticles their characterization and biomedical application. Adv Nat Sci Nanosci Nanotechnol. 2012; 3:015013.
• [30]Nowack B, Krug HF, Height M. 120 years of nanosilver history: implications for policy makers. Environ Sci Technol. 2011; 45:1177-1183.
• [31]Schacht VJ, Neumann LV, Sandhi SK, Chen L, Henning T, Klar PJ, Theophel K, Schnell S, Bunge M. Effects of silver nanoparticles on microbial growth dynamics. J Appl Microbiol. 2013; 114:25-35.
• [32]Rai M, Yadav A, Gade A. Silver nanoparticles as a new generation of antimicrobials. Biotechnol Adv. 2009; 27:76-83.
• [33]Zhao GJ, Stevens SE. Multiple parameters for the comprehensive evaluation of the susceptibility of Escherichia coli to the silver ion. Biometals. 1998; 11:27-32.
• [34]Loo CY, Young PM, Lee WH, Cavaliere R, Whitchurch CB, Rohanizadeh R. Non-cytotoxic silver nanoparticle-polyvinyl alcohol hydrogels with anti-biofilm activity: designed as coatings for endotracheal tube materials. Biofouling. 2014; 30:773-788.
• [35]Rai MK, Deshmukh SD, Ingle AP, Gade AK. Silver nanoparticles: the powerful nanoweapon against multidrug-resistant bacteria. J Appl Microbiol. 2012; 112:841-852.
• [36]Stoimenov PK, Klinger RL, Marchin GL, Klabunde KJ. Metal oxide nanoparticles as bactericidal agents. Langmuir. 2002; 18:6679-6686.
• [37]Sondi I, Salopek-Sondi B. Silver nanoparticles as antimicrobial agent: a case study on E-coli as a model for Gram-negative bacteria. J Colloid Interface Sci. 2004; 275:177-182.
• [38]Benhabiles S, Salah R, Lounici H, Drouiche N, Goosen MFA, Mameri N. Antibacterial activity of chitin, chitosan and its oligomers prepared from shrimp shell waste. Food Hydrocolloids. 2012; 29:48-56.
• [39]Sanpui P, Murugadoss A, Prasad PVD, Ghosh SS, Chattopadhyay A. The antibacterial properties of a novel chitosan-Ag-nanoparticle composite. Int J Food Microbiol. 2008; 124:142-146.
• [40]Dawy M, Rifaat HM, Moustafa SA, Mousa HA. Physicochemical studies on nano silver particles preparated by different techniques. Aust J Basic Appl Sci. 2012; 6:257-262.
• [41]Lok CN, Ho CM, Chen R, He QY, Yu WY, Sun H, Tam PKH, Chiu JF, Che CM. Silver nanoparticles: partial oxidation and antibacterial activities. J Biol Inorg Chem. 2007; 12:527-534.
• [42]Kim JS, Kuk E, Yu KN, Kim JH, Park SJ, Lee HJ, Kim SH, Park YK, Park YH, Hwang CY et al.. Antimicrobial effects of silver nanoparticles. Nanomed Nanotechnol Biol Med. 2007; 3:95-101.
• [43]Egger S, Lehmann RP, Height MJ, Loessner MJ, Schuppler M. Antimicrobial properties of a novel silver-silica nanocomposite material. Appl Environ Microbiol. 2009; 75:2973-2976.
• [44]Singh K, Panghal M, Kadyan S, Chaudhary U, Yadav JP. Antibacterial activity of synthesized silver nanoparticles from Tinospora cordifolia against multi drug resistant strains of Pseudomonas aeruginosa isolated from burn patients. J Nanomed Nanotechnol. 2014; 5:192.
• [45]Khanna PK, Singh N, Charan S, Subbarao V, Gokhale R, Mulik UP. Synthesis and characterization of Ag/PVA nanocomposite by chemical reduction method. Mater Chem Phys. 2005; 93:117-121.
• [46]Lee PC, Meisel D. Adsorption and surface-enhanced Raman of dyes on silver and gold sols. J Phys Chem. 1982; 86:3391-3395.
• [47]Temgire MK, Joshi SS. Optical and structural studies of silver nanoparticles. Radiat Phys Chem. 2004; 71:1039-1044.
• [48]Cao XL, Cheng C, Ma YL, Zhao CS. Preparation of silver nanoparticles with antimicrobial activities and the researches of their biocompatibilities. J Mat Sci Mat Med. 2010; 21:2861-2868.
• [49]Shih CM, Shieh YT, Twu YK. Preparation of gold nanopowders and nanoparticles using chitosan suspensions. Carbohydr Polym. 2009; 78:309-315.
• [50]Hindler J. Tests to assess bactericidal activity. Clinical microbiology procedures handbook. Isenberg HD, editor. American Society for Microbiology, Washington, D.C.; 1992.
• [51]Leite AM, Lima EO, Souza EL, Diniz MF, TrajanoII VN, Medeiros IA. Inhibitory effect of b-pinene, a-pinene and eugenol on the growth of potential infectious endocarditis causing Gram-positive bacteria. Revista Brasileira de Ciências Farmacêuticas. 2007; 43:121-126.
• [52]GiamarellosBouroulis EJ, Grecka P, Giamarellou H. Comparative in vitro interactions of ceftazidime, meropenem, and imipenem with amikacin on multiresistant Pseudomonas aeruginosa. Diagn Microbiol Infect Dis. 1997; 29:81-86.