In Vitro and in Vivo Antibacterial Activities of Cyanidinum Chloride-loaded Liposomes against a Resistant Strain of Pseudomonas aeruginosa

Abstract

Pseudomonas aeruginosa remains a common cause of wound infections. Different studies have shown that the entrapment of plant-derived materials in liposomes could increase their antibacterial activity against Pseudomonas aeruginosa. The aim of this study was to prepare cyanidinum chloride-loaded liposomes and evaluate their in vitro and in vivo efficacy against a resistant strain of Pseudomonas aeruginosa ATCC 15692. Cyanidinum chloride-loaded liposomes were prepared by extrusion method. The minimum inhibitory concentrations of cyanidinum chloride in the free and liposomal forms against Pseudomonas aeruginosa ATCC 15 692 were determined in vitro by broth dilution method. The in vitro killing rates for free and liposomal cyanidinum chloride were analyzed. Ultimately, the in vivo therapeutic efficacy of the prepared liposomes in mice skin infected by ATCC 15692 was investigated. The minimum inhibitory concentrations of the free and liposomal forms of cyanidinum chloride against ATCC 15692 were 1.5 × 10⁻³ and 7.7 × 10⁻⁴ M, respectively. In vivo treatment with the free and cyanidinum chloride-loaded liposomes resulted in almost 40 and 100 % survival rates, respectively. Our results showed that cyanidinum chloride-loaded liposomes would be a good choice for the treatment of wound infection caused by Pseudomonas aeruginosa because of their high effectiveness.

Key words

cyanidinum chloride - liposome - Pseudomonas aeruginosa - killing rates - in vitro - in vivo

• References

• 1 Plotnikova JM, Rahme LG, Ausubel FM. Pathogenesis of the human opportunistic pathogen Pseudomonas aeruginosa PA14 in Arabidopsis . Plant Physiol 2000; 124: 1766-1774
• 2 Buijs J, Dofferhoff AS, Mouton JW, van der Meer JW. Continuous administration of PBP-2- and PBP-3-specific beta-lactams causes higher cytokine responses in murine Pseudomonas aeruginosa and Escherichia coli sepsis. J Antimicrob Chemother 2007; 59: 926-933
• 3 Gibbons S. Phytochemicals for bacterial resistance-strengths, weaknesses and opportunities. Planta Med 2008; 74: 594-602
• 4 Kulling SE, Rawel HM. Chokeberry (Aronia melanocarpa) – A review on the characteristic components and potential health effects. Planta Med 2008; 74: 1625-1634
• 5 Prasad S, Phromnoi K, Yadav VR, Chaturvedi MM, Aggarwal BB. Targeting inflammatory pathways by flavonoids for prevention and treatment of cancer. Planta Med 2010; 76: 1044-1063
• 6 Truong VD, Hua Z, Thompson RL, Yencho GC, Pecota KV. Pressurized liquid extraction and quantification of anthocyanins in purple-fleshed sweet potato genotypes. J Food Compost Anal 2012; 26: 96-103
• 7 Truong VD, Deighton N, Thompson RT, McFeeters RF, Dean LO, Pecota KV, Yencho GC. Characterization of anthocyanins and anthocyanidins in purple-fleshed sweetpotatoes by HPLC-DAD/ESI-MS/MS. J Agric Food Chem 2010; 58: 404-410
• 8 Naz S, Siddiqi R, Ahmad S, Rasool SA, Sayeed SA. Antibacterial activity directed isolation of compounds from Punica granatum . J Food Sci 2007; 72: 341-345
• 9 Cushnie TP, Lamb AJ. Antimicrobial activity of flavonoids. Int J Antimicrob Agents 2005; 26: 343-356
• 10 Hassine DB, Abderrabba M, Yvon Y, Lebrihi A, Mathieu F, Couderc F, Bouajila J. Chemical composition and in vitro evaluation of the antioxidant and antimicrobial activities of Eucalyptus gillii essential oil and extracts. Molecules 2012; 17: 9540-9558
• 11 Khlifi D, Hamdi M, El Hayouni A, Cazaux S, Souchard JP, Couderc F, Bouajila J. Global chemical composition and antioxidant and anti-tuberculosis activities of various extracts of Globularia alypum L. (Globulariaceae) leaves. Molecules 2011; 16: 10592-10603
• 12 Safinya CR, Ewert KK, Leal C. Cationic liposome-nucleic acid complexes: liquid crystal phases with applications in gene therapy. Liq Cryst 2011; 38: 1715-1723
• 13 Heneweer C, Gendy SE, Peñate-Medina O. Liposomes and inorganic nanoparticles for drug delivery and cancer imaging. Ther Deliv 2012; 3: 645-656
• 14 Ellbogen MH, Olsen KM, Gentry-Nielsen MJ, Preheim LC. Efficacy of lipo-some-encapsulated ciprofloxacin compared with ciprofloxacin and ceftriaxone in a rat model of pneumococcal pneumonia. J Antimicrob Chemother 2003; 51: 83-91
• 15 Zhong Z, Wan Y, Han J, Shi S, Zhang Z, Sun X. Improvement of adenoviral vector-mediated gene transfer to airway epithelia by folate-modified anionic liposomes. Int J Nanomed 2011; 6: 1083-1093
• 16 Roychoudhury J, Sinha R, Ali N. Therapy with sodium stibogluconate in stearylamine-bearing liposomes confers cure against SSG-resistant Leishmania donovani in BALB/c mice. PLoS One 2011; 6: e17376
• 17 Fang JY, Hwang TL, Huang YL, Fang CL. Enhancement of the transdermal delivery of catechins by liposomes incorporating anionic surfactants and ethanol. Int J Pharm 2006; 310: 131-138
• 18 Youfang J, Fenghung C, Longhwang T, Linghuang Y. Physicochemical characteristics and in vivo deposition of liposome-encapsulated tea catechins by topical and intratumor administrations. J Drug Targ 2005; 13: 19-27
• 19 Nunes PS, Albuquerque Jr. RL, Cavalcante DR, Dantas MD, Cardoso JC, Bezerra MS, Souza JC, Serafini MR, Quitans Jr. LJ, Bonjardim LR, Araújo AA. Collagen-based films containing liposome-loaded usnic acid as dressing for dermal burn healing. J Biomed Biotechnol 2011; 2011: 761593
• 20 Huang CM, Chen CH, Pornpattananangkul D, Zhang L, Chan M, Hsieh MF, Zhang L. Eradication of drug resistant Staphylococcus aureus by liposomal oleic acids. Biomaterials 2011; 32: 214-221
• 21 Committe on the Care and Use of Laboratory Animals of the Institute of Laboratory Animal Resources, Commission on Life Sciences National Research Council. Guide for the care and use of laboratory animals. Washington, D.C.: National Academy Press; 1996
• 22 Geetha M, Saravanakumar M, Suganya Devi P. HPLC analysis of quercetin and cyanidin from onion peel (Allium cepa L.). Am J Pharm Tech Res 2012; 2: 331-337
• 23 Gharib A, Faezizadeh Z, Mohammad Asghari H. Preparation and antifungal activity of spray-dried amphotericin B-loaded nanospheres. Daru 2011; 19: 351-355
• 24 Cabral ECM, Zollner RL, Santana MHA. Preparation and characterization of liposomes entrapping allergenic proteins. Braz J Chem Eng 2004; 21: 137-146
• 25 National Committee for Clinical Laboratory Standards. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically: approved standard, 6th edition. Wayne: National Committee for Clinical Laboratory Standards; 2003. NCCLS document M7-A6
• 26 Shrivastava SM, Saurabh S, Rai D, Dwivedi VK, Chaudhary M. In vitro microbial efficacy of sulbactomax: a novel fixed dose combination of ceftriaxone sulbactam and ceftriaxone alone. Curr Drug Ther 2009; 4: 73-77
• 27 Cao S, Sun LQ, Wang M. Antimicrobial activity and mechanism of action of Nu-3, a protonated modified nucleotide. Ann Clin Microbiol Antimicrob 2011; 10: 1-9
• 28 Mitkari BV, Korde SA, Mahadik KR, Kokare CR. Formulation and evaluation of topical liposomal gel for fluconazole. Indian J Educ Res 2010; 44: 324-333
• 29 van Vuuren SF, du Toit LC, Parry A, Pillay V, Choonara YE. Encapsulation of essential oils within a polymeric liposomal formulation for enhancement of antimicrobial efficacy. Nat Prod Commun 2010; 5: 1401-1408
• 30 van Vuuren S, Viljoen A. Plant-based antimicrobial studies-methods and approaches to study the interaction between natural products. Planta Med 2011; 77: 1168-1182
• 31 Muqbil I, Masood A, Sarkar FH, Mohammad RM, Azmi AS. Progress in nanotechnology based approaches to enhance the potential of chemopreventive agents. Cancers 2011; 3: 428-445
• 32 Shaffa MW, Dayem SA, Elshemy WM. In vitro antibacterial activity of liposomal cephalexin against Staphylococcus aureus . Romanian J Biophys 2008; 18: 293-300
• 33 Detoni CB, Cabral-Albuquerque ECM, Hohlemweger SVA, Sampaio C, Barros TF, Velozo EF. Essential oil from Zanthoxylum tingoassuiba loaded into multilamellar liposomes useful as antimicrobial agents. J Microencapsul 2009; 26: 684-691
• 34 Klein DG, Fritsh DE, Amin SG. Wound infection following trauma and burn injuries. Crit Care Nursing N Am 1995; 7: 627-642
• 35 Pruitt Jr. BA, McManus AT, Kim SH, Goodwin CW. Burn wound infections: current status. World J Surg 1998; 22: 135-145
• 36 Wurtz R, Karajovic E, Dacumos E, Hanumandass M. Nosocomial infections in a burn intensive care unit. Burns 1995; 21: 180-184
• 37 Karodi R, Jadhav M, Rub R, Bafna A. Evaluation of the wound healing activity of a crude extract of Rubia cordifolia L. (Indian madder) in mice. Int J Appl Res Nat Prod 2009; 2: 12-18