https://doi.org/10.5812/pedinfect.21481

Comparison of Difference Between Fluconazole and Silver Nanoparticles in Antimicrobial Effect on Fluconazole-Resistant Candida Albicans Strains

authors:

avatar Shadi Alimehr 1 , avatar Hamide Shekari Ebrahim Abad 2 , avatar Ahmadreza Shahverdi 3 , avatar Jamal Hashemi 1 , avatar Kamyar Zomorodian 4 , avatar Maryam Moazeni 5 , avatar Sahar Vosoghian 2 , avatar Sassan Rezaie 1 , *

Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran
Pediatric Infections Research Center, Mofid Children’s Hospital, Shahid Beheshti University of Medical Sciences, Tehran, IR Iran
Faculty of Pharmacy, Pharmaceutical Sciences Research Center, Tehran University of Medical Sciences, Tehran, IR Iran
Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, IR Iran
Department of Medical Mycology, School of Medicine, Mazandaran University of Medical Sciences, Sari, IR Iran
Archives of Pediatric Infectious Diseases: Vol.3, issue 2; e21481
published online: April 15, 2015
article type: Research Article
received: June 23, 2014
revised: December 3, 2014
accepted: January 28, 2015

how to cite: Alimehr S, Shekari Ebrahim Abad H, Shahverdi A, Hashemi J, Zomorodian K, et al. Comparison of Difference Between Fluconazole and Silver Nanoparticles in Antimicrobial Effect on Fluconazole-Resistant Candida Albicans Strains. Arch Pediatr Infect Dis. 2015;3(2):e21481. https://doi.org/10.5812/pedinfect.21481.

Abstract

Background:

Opportunistic fungi cause fungal infections. Whereas some microorganisms are resistant to chemical drugs, scientists are looking for new natural and inorganic antimicrobial agents. The recent research on metal nanoparticles showed that silver nanoparticles (nanosilver) exhibits lower toxicity to mammalian cells and higher toxicity to microorganisms.

Objectives:

This study aimed to compare the difference between antimicrobial effect of nanosilver and some antibiotic agents on Candidaalbicans.

Materials and Methods:

We studied effect of fluconazole, nanosilver, and their combination on 20 fluconazole-resistant C. albicans from two centers and one standard sample (ATCC10261) by minimal inhibitory concentration (MIC) method.

Results:

Result of fungi static and fungicidal activities of nanosilver plus fluconazole on fluconazole-resistant C. albicans showed better inhibitory effect on the growth of standard C. albicans when MIC of fluconazole (8 µg/mL) combined with MIC of Nanosilver (0.0625 µg/mL).

Conclusions:

Totally, our results showed nanosilver caused an increase of at least nine-fold in inhibitory effect of fluconazole.

1. Background

In recent years, morbidity and mortality are increased significantly by severe fungal infections (1). Candida species have been one of the most common pathogens responsible for fungal infections, which cause hospital-acquired sepsis with annually mortality rate of up to 40% (2). Opportunistic fungi cause fungal infections, especially in vulnerable people with special conditions such as pregnancy or HIV-positive and immune-compromised patients who need intensive treatment with broad-spectrum antibiotics (3-5). Nowadays most of the available effective antifungal agents are based on polyenes (amphotericin B), echinocandins (caspofungin, micafungin, and anidulafungin) and triazoles (fluconazole, itraconazole, voriconazole, and posaconazole) (6, 7). However, scientists are looking for new natural and inorganic antimicrobial agents (8, 9). The recent research on metal nanoparticles showed that silver nanoparticles (nanosilvers) have received special attention as a possible antimicrobial agent (10-16). Since ancient times, silver has been used widely to treat infections and has strong inhibitory effects as well as a broad spectrum of antimicrobial activities against microorganisms, which has been thoroughly investigated (1, 9, 15, 17). This toxicity effect on bacteria has been investigated for more than 60 years (16) and in comparison to other metals, silver exhibits lower toxicity to mammalian cells and higher toxicity to microorganisms (18). Nanosilver exerts antimicrobial effects through interacting with main components of microorganisms including DNA (19), microbial proteins (20), and cell wall (20, 21); moreover, nanosilver produces reactive oxygen species (ROS) (21). The accumulation of intracellular ROS is as an important regulator for starting early apoptosis phase (22). Subsequently, increasing level of intracellular ROS lead to initiation of mitochondrial fragmentation (23).

2. Objectives

Regarding comparison of difference between antimicrobial effect of nanosilver and some antibiotic agents on Candida albicans, we compared the effect of nanosilver with fluconazole and their combination on collected fluconazole-resistant and fluconazole-sensitive C.albicans.

4. Results

Result of fungi static and fungicidal activities of fluconazole against C. albicans showed: 1) The MIC of fluconazole concentration for standard sample was 16 µg/mL (Table 1). 2) The growth of 20 fluconazole-resistant C. albicans was inhibited at MICs > 512 µg/mL (Table 1).

Result of fungi static and fungicidal activities of nanosilver against C. albicans showed: 1) The MIC of nanosilver for standard sample was 4 µg/mL. (Table 2). 2) The MICs of nanosilver for 20 resistant C. albicans were 2 µg/mL (58%) and 4 µg/mL (42%).

Result of fungi static and fungicidal activities of nanosilver plus fluconazole (8 µg/mL) on C. albicans showed: 1) the combination had better inhibitory effect on the growth of standard C. albicans when MIC of fluconazole (8 µg/mL) was combined with MIC of Nanosilver (0.0625 µg/mL) (Table 3). 2) Results on 20 resistant C. albicans showed there are several MIC of nanosilver: 40% of resistant C. albicans samples grew on 0.25 µg/mL, 11% on 0.0625 µg/mL, 22% on 0.03125 µg/mL, and 27% had no growth on 0.03125 (Table 3).

Table 1.

The Growths of Standard and Fluconazole-Resistant Candida albicans on Difference Fluconazole Concentrations

VariablesFluconazole Concentrations, µg/mL
0.51248163264128256512
ResistantCandida albicans samples+++++++++++
Standard sample+++++------
Table 2.

The Growths of Standard Sample and Fluconazole-Resistant Candida albicans on Difference Nanosilver Concentrations

VariablesNanosilver Concentrations, µg/mL
0.51248163264
58% of resistantCandida albicans samples +-------
42% of resistant Candida albicans samples ++------
Standard sample+++-----
Table 3.

The Growth of Standard and Fluconazole-resistant Candida albicans on Difference Nanosilver Concentrations Combined with 8 µg/mL Fluconazole

VariablesNanosilver Concentrations (0.5-0.0625 µg/mL) Plus 8 µg/mL of Fluconazole
0.0156250.031250.06250.1250.25
22% of resistant Candida albicans samples ++---
11% of resistant Candidaalbicans samples +++--
40% of resistant Candida albicans samples +++++
27% of resistant Candida albicans samples +----
Standard sample+----

References

  • 1.

    Pfaller MA, Diekema DJ. Epidemiology of invasive candidiasis: a persistent public health problem. Clin Microbiol Rev. 2007;20(1):133-63. [PubMed ID: 17223626]. https://doi.org/10.1128/CMR.00029-06.

  • 2.

    Patterson TF. Treatment and prevention of fungal infections. Focus on candidemia. New York: Applied Clinical Education; 2007.

  • 3.

    Nasrollahi A, Pourshamsian K, Mansourkiaee P. Antifungal activity of silver nanoparticles on some of fungi. Int J Nano Dim. 2011;1(3):233-9.

  • 4.

    Martin GS, Mannino DM, Eaton S, Moss M. The epidemiology of sepsis in the United States from 1979 through 2000. N Engl J Med. 2003;348(16):1546-54. [PubMed ID: 12700374]. https://doi.org/10.1056/NEJMoa022139.

  • 5.

    Pappas PG, Rex JH, Lee J, Hamill RJ, Larsen RA, Powderly W, et al. A prospective observational study of candidemia: epidemiology, therapy, and influences on mortality in hospitalized adult and pediatric patients. Clin Infect Dis. 2003;37(5):634-43. [PubMed ID: 12942393]. https://doi.org/10.1086/376906.

  • 6.

    Levin MD, den Hollander JG, van der Holt B, Rijnders BJ, van Vliet M, Sonneveld P, et al. Hepatotoxicity of oral and intravenous voriconazole in relation to cytochrome P450 polymorphisms. J Antimicrob Chemother. 2007;60(5):1104-7. [PubMed ID: 17827141]. https://doi.org/10.1093/jac/dkm330.

  • 7.

    Venkatakrishnan K, von Moltke LL, Greenblatt DJ. Effects of the antifungal agents on oxidative drug metabolism: clinical relevance. Clin Pharmacokinet. 2000;38(2):111-80. [PubMed ID: 10709776]. https://doi.org/10.2165/00003088-200038020-00002.

  • 8.

    Kim TN, Feng QL, Kim JO, Wu J, Wang H, Chen GC, et al. Antimicrobial effects of metal ions (Ag+, Cu2+, Zn2+) in hydroxyapatite. J Mater Sci Mater Med. 1998;9(3):129-34. [PubMed ID: 15348901].

  • 9.

    Cho KH, Park JE, Osaka T, Park SG. The study of antimicrobial activity and preservative effects of nanosilver ingredient. Electrochimica Acta. 2005;51(5):956-60. https://doi.org/10.1016/j.electacta.2005.04.071.

  • 10.

    Silver S. Bacterial silver resistance: molecular biology and uses and misuses of silver compounds. FEMS Microbiol Rev. 2003;27(2-3):341-53. [PubMed ID: 12829274].

  • 11.

    Baker C, Pradhan A, Pakstis L, Pochan DJ, Shah SI. Synthesis and antibacterial properties of silver nanoparticles. J Nanosci Nanotechnol. 2005;5(2):244-9. [PubMed ID: 15853142].

  • 12.

    Lee BU, Yun SH, Ji JH, Bae GN. Inactivation of S. epidermidis, B. subtilis, and E. coli bacteria bioaerosols deposited on a filter utilizing airborne silver nanoparticles. J Microbiol Biotechnol. 2008;18(1):176-82. [PubMed ID: 18239437].

  • 13.

    Melaiye A, Sun Z, Hindi K, Milsted A, Ely D, Reneker DH, et al. Silver(I)-imidazole cyclophane gem-diol complexes encapsulated by electrospun tecophilic nanofibers: formation of nanosilver particles and antimicrobial activity. J Am Chem Soc. 2005;127(7):2285-91. [PubMed ID: 15713108]. https://doi.org/10.1021/ja040226s.

  • 14.

    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(1):177-82. [PubMed ID: 15158396]. https://doi.org/10.1016/j.jcis.2004.02.012.

  • 15.

    Lok CN, Ho CM, Chen R, He QY, Yu WY, Sun H, et al. Proteomic analysis of the mode of antibacterial action of silver nanoparticles. J Proteome Res. 2006;5(4):916-24. [PubMed ID: 16602699]. https://doi.org/10.1021/pr0504079.

  • 16.

    Franke S, Grass G, Nies DH. The product of the ybdE gene of the Escherichia coli chromosome is involved in detoxification of silver ions. Microbiology. 2001;147(Pt 4):965-72. [PubMed ID: 11283292].

  • 17.

    Kim JS, Kuk E, Yu KN, Kim JH, Park SJ, Lee HJ, et al. Antimicrobial effects of silver nanoparticles. Nanomedicine. 2007;3(1):95-101. [PubMed ID: 17379174]. https://doi.org/10.1016/j.nano.2006.12.001.

  • 18.

    Zhao G, Stevens SJ. Multiple parameters for the comprehensive evaluation of the susceptibility of Escherichia coli to the silver ion. Biometals. 1998;11(1):27-32. [PubMed ID: 9450315].

  • 19.

    Yang W, Shen C, Ji Q, An H, Wang J, Liu Q, et al. Food storage material silver nanoparticles interfere with DNA replication fidelity and bind with DNA. Nanotechnology. 2009;20(8):85102. [PubMed ID: 19417438]. https://doi.org/10.1088/0957-4484/20/8/085102.

  • 20.

    Yamanaka M, Hara K, Kudo J. Bactericidal actions of a silver ion solution on Escherichia coli, studied by energy-filtering transmission electron microscopy and proteomic analysis. Appl Environ Microbiol. 2005;71(11):7589-93. [PubMed ID: 16269810]. https://doi.org/10.1128/AEM.71.11.7589-7593.2005.

  • 21.

    Park HJ, Kim JY, Kim J, Lee JH, Hahn JS, Gu MB, et al. Silver-ion-mediated reactive oxygen species generation affecting bactericidal activity. Water Res. 2009;43(4):1027-32. [PubMed ID: 19073336]. https://doi.org/10.1016/j.watres.2008.12.002.

  • 22.

    Benaroudj N, Lee DH, Goldberg AL. Trehalose accumulation during cellular stress protects cells and cellular proteins from damage by oxygen radicals. J Biol Chem. 2001;276(26):24261-7. [PubMed ID: 11301331]. https://doi.org/10.1074/jbc.M101487200.

  • 23.

    Pozniakovsky AI, Knorre DA, Markova OV, Hyman AA, Skulachev VP, Severin FF. Role of mitochondria in the pheromone- and amiodarone-induced programmed death of yeast. J Cell Biol. 2005;168(2):257-69. [PubMed ID: 15657396]. https://doi.org/10.1083/jcb.200408145.

  • 24.

    Goa KL, Barradell LB. Fluconazole. An update of its pharmacodynamic and pharmacokinetic properties and therapeutic use in major superficial and systemic mycoses in immunocompromised patients. Drugs. 1995;50(4):658-90. [PubMed ID: 8536553].

  • 25.

    Arevalo MP, Arias A, Andreu A, Rodriguez C, Sierra A. Fluconazole, itraconazole and ketoconazole in vitro activity against Candida spp. J Chemother. 1994;6(4):226-9. [PubMed ID: 7830098].

  • 26.

    Van't Wout JW. Fluconazole treatment of candidal infections caused by non-albicans Candida Species. Eur J Clin Microbiol. 1996;15:228-42.

  • 27.

    Furno F, Morley KS, Wong B, Sharp BL, Arnold PL, Howdle SM, et al. Silver nanoparticles and polymeric medical devices: a new approach to prevention of infection? J Antimicrob Chemother. 2004;54(6):1019-24. [PubMed ID: 15537697]. https://doi.org/10.1093/jac/dkh478.

  • 28.

    Pfaller MA, Diekema DJ, Sheehan DJ. Interpretive breakpoints for fluconazole and Candida revisited: a blueprint for the future of antifungal susceptibility testing. Clin Microbiol Rev. 2006;19(2):435-47. [PubMed ID: 16614256]. https://doi.org/10.1128/CMR.19.2.435-447.2006.

  • 29.

    Enwuru CA, Ogunledun A, Idika N, Enwuru NV, Ogbonna F, Aniedobe M, et al. Fluconazole resistant opportunistic oro-pharyngeal Candida and non-Candida yeast-like isolates from HIV infected patients attending ARV clinics in Lagos, Nigeria. Afr Health Sci. 2008;8(3):142-8. [PubMed ID: 19357740].

  • 30.

    Kim KJ, Sung WS, Moon SK, Choi JS, Kim JG, Lee DG. Antifungal effect of silver nanoparticles on dermatophytes. J Microbiol Biotechnol. 2008;18(8):1482-4. [PubMed ID: 18756112].

  • 31.

    Kvitek L, Panacek A, Prucek R, Soukupova J, Vanickova M, Kolar M, et al. Antibacterial activity and toxicity of silver – nanosilver versus ionic silver. J Phys: Conference Series. 2011;304:12029. https://doi.org/10.1088/1742-6596/304/1/012029.