Research Article
Efficacy of Trans-cinnamaldehyde and Eugenol in Reducing Acinetobacter baumannii Adhesion to and Invasion of Human Keratinocytes and Controlling Wound Infection In Vitro
Deepti P. Karumathil,
Meera Surendran-Nair,
Kumar Venkitanarayanan,
Deepti P. Karumathil
Department of Animal Science, University of Connecticut, Storrs, CT, 06269 USA
Search for more papers by this authorMeera Surendran-Nair
Department of Animal Science, University of Connecticut, Storrs, CT, 06269 USA
Search for more papers by this authorCorresponding Author
Kumar Venkitanarayanan
Department of Animal Science, University of Connecticut, Storrs, CT, 06269 USA
Correspondence to: Kumar Venkitanarayanan, Department of Animal Science, University of Connecticut, Storrs, CT 06269, USA.
E-mail: [email protected]
Search for more papers by this authorDeepti P. Karumathil,
Meera Surendran-Nair,
Kumar Venkitanarayanan,
Deepti P. Karumathil
Department of Animal Science, University of Connecticut, Storrs, CT, 06269 USA
Search for more papers by this authorMeera Surendran-Nair
Department of Animal Science, University of Connecticut, Storrs, CT, 06269 USA
Search for more papers by this authorCorresponding Author
Kumar Venkitanarayanan
Department of Animal Science, University of Connecticut, Storrs, CT, 06269 USA
Correspondence to: Kumar Venkitanarayanan, Department of Animal Science, University of Connecticut, Storrs, CT 06269, USA.
E-mail: [email protected]
Search for more papers by this authorAbstract
The study investigated the efficacy of two natural, plant-derived antimicrobials (PDAs), namely trans-cinnamaldehyde (TC), and eugenol (EG) for decreasing Acinetobacter baumannii adhesion to and invasion of human keratinocytes (HEK001). Moreover, the efficacy of two PDAs for inhibiting A. baumannii biofilm formation was determined using an in vitro collagen matrix wound model. Additionally, the effect of TC and EG on A. baumannii biofilm architecture was visualized using confocal scanning microscopy. Further the effect of both PDAs on genes critical for biofilm synthesis was determined using real-time quantitative polymerase chain reaction. Both TC and EG significantly reduced A. baumannii adhesion and invasion to HEK001 by ~2 to 3 log10 CFU/mL (p < 0.05) compared with the controls (p < 0.05). Further, after 24 and 48 h, TC and EG inhibited biofilm formation by ~1.5 to 2 and ~2 to 3.5 log10 CFU/mL, compared with controls (p < 0.05). Confocal microscopy revealed that TC and EG disrupted the biofilm architecture. RT-qPCR results indicated that two phytochemicals significantly down-regulated the transcription of genes associated with A. baumannii biofilm production. The results suggest that both TC and EG could potentially be used to treat A. baumannii wound infections; however, their efficacy in in vivo models needs to be validated. Copyright © 2016 John Wiley & Sons, Ltd.
References
- Ali A, Botha J, Tiruvoipati R. 2014. Fatal skin and soft tissue infection of multidrug resistant Acinetobacter baumannii: a case report. Int J Surg Case Rep 5(8): 532–6.
- Altiok D, Altiok E, Tihminlioglu F. 2010. Physical, antibacterial and antioxidant properties of chitosan films incorporated with thyme oil for potential wound healing applications. J Mater Sci Mater Med 21(7): 2227–36.
- Amalaradjou MA, Narayanan A, Baskaran SA, Venkitanarayanan K. 2010. Antibiofilm effect of trans-cinnamaldehyde on uropathogenic Escherichia coli. J Urol31; 184(1): 358–63.
- Amalaradjou MA, Venkitanarayanan K. 2011. Effect of trans-cinnamaldehyde on reducing resistance to environmental stresses in Cronobacter sakazakii. Foodborne Pathog Dis 8(3): 403–9.
- Aoyagi S, Onishi H, Machida Y. 2007. Novel chitosan wound dressing loaded with minocycline for the treatment of severe burn wounds. Int J Pharm 330(1): 138–45.
- Athanasopoulos AN, Economopoulou M, Orlova VV, et al. 2006. The extracellular adherence protein (Eap) of Staphylococcus aureus inhibits wound healing by interfering with host defense and repair mechanisms. Blood 107: 2720–2727.
- Bergogne-Berezin E, Towner KJ. 1996. Acinetobacter spp. as nosocomial pathogens: microbiological, clinical, and epidemiological features. Clin Microbiol Rev 9: 148–165.
- Boateng JS, Pawar HV, Tetteh J. 2013. Polyox and carrageenan based composite film dressing containing anti-microbial and anti-inflammatory drugs for effective wound healing. Int J Pharm 441: 181–191.
- Boateng J, Catanzano O. 2015. Advanced therapeutic dressings for effective wound healing—a review. J Pharm Sci 104: 3653–3680.
- Bookout AL, Mangelsdorf DJ. 2003. Quantitative real-time PCR protocol for analysis of nuclear receptor signaling pathways. Nucl Recept Signal 1: e012.
- Brossard KA, Campagnari AA. 2012. The Acinetobacter baumannii biofilm-associated protein plays a role in adherence to human epithelial cells. Infect Immun 80(1): 228–233.
- Brooks SE, Walczak MA, Hameed R, Coonan P. 2002. Chlorhexidine resistance in antibiotic-resistant bacteria isolated from the surfaces of dispensers of soap containing chlorhexidine. Infect Control Hosp Epidemiol 23: 692–695.
- Carson CF, Mee BJ, Riley TV. 2002. Mechanism of action of Melaleuca alternifolia (tea tree) oil on Staphylococcus aureus determined by time-kill, lysis, leakage and salt tolerance assays and electron microscopy. Antimicrob Agents Chemother 46(6): 1914–1920.
- Cox SD, Mann CM, Markham JL, et al. 2000. The mode of antimicrobial action of the essential oil of Melaleuca alternifolia (tea tree oil). J Appl Microbiol 88(1): 170–175.
- Dai T, Huang YY, Sharma SK, Hashmi JT, Kurup DB, Hamblin MR. 2010. Topical antimicrobials for burn wound infections. Recent Pat Antiinfect Drug Discov 5: 124–151.
- Daker M, Lin VY, Akowuah GA, Yam MF, Ahmad M. 2013. Inhibitory effects of Blume stem bark extract and cinnamaldehyde on nasopharyngeal carcinoma cells; synergism with cisplatin. Exp Ther Med 5: 1701–1709.
- Dallo SF, Weitao T. 2010. Insights into Acinetobacter war-wound infections, biofilms, and control. Adv Skin Wound Care 23: 169–174.
- de Breij A, Haisma EM, Rietveld M, et al. 2012. Three-dimensional human skin equivalent as a tool to study Acinetobacter baumannii colonization. Antimicrob Agents Chemother 56: 2459–2464.
- Djeraba A, Quere P. 2000. In vivo macrophage activation in chickens with Acemannan, a complex carbohydrate extracted from aloe vera. Int J Immunopharmacol 22: 365–372.
- Dorsey CW, Tomaras AP, Actis LA. 2002. Genetic and phenotypic analysis of Acinetobacter baumannii insertion derivatives generated with a transposome system. Appl Environ Microbiol 68(12): 6353–6360.
- Durante-Mangoni E, Zarrilli R. 2011. Global spread of drug-resistant Acinetobacter baumannii: molecular epidemiology and management of antimicrobial resistance. Future Microbiol 6: 407–422.
- Dursun N, Liman N, Özyazgan I, Günes I, Saraymen R. 2003. Role of thymus oil in burn wound healing. J Burn Care Res 24: 395–399.
- Esterly JS, Griffith M, Qi C, Malczynski M, Postelnick MJ, Scheetz MH. 2011. Impact of carbapenem resistance and receipt of active antimicrobial therapy on clinical outcomes of Acinetobacter baumannii bloodstream infections. Antimicrob Agents Chemother 5: 4844–4849.
- Evans HL, Dellit TH, Chan J, Nathens AB, Maier RV, Cuschieri J. 2010. Effect of chlorhexidine whole-body bathing on hospital-acquired infections among trauma patients. Arch Surg(Chicago, Ill.: 1960) 145: 240–246.
- Farahpour MR, Habibi M. 2012. Evaluation of the wound healing activity of an ethanolic extract of Ceylon cinnamon in mice. Vet Med 57: 53–57.
- Gaddy JA, Actis LA. 2009. Regulation of Acinetobacter baumannii biofilm formation.
- Gaddy JA, Tomaras AP, Actis LA. 2009. The Acinetobacter baumannii 19606 OmpA protein plays a role in biofilm formation on abiotic surfaces and in the interaction of this pathogen with eukaryotic cells. Infect Immun 77: 3150–3160.
- Giamarellou H, Antoniadou A, Kanellakopoulou K. 2008. Acinetobacter baumannii: a universal threat to public health? Int J Antimicrob 32: 106–119.
- Gill AO, Holley RA. 2004. Mechanisms of bactericidal action of cinnamaldehyde against Listeria monocytogenes and of eugenol against L. monocytogenes and Lactobacillus sakei. Appl Environ Microbiol 70: 5750–5755.
- Guerrero DM, Perez F, Conger NG, et al. 2010. Acinetobacter baumannii-associated skin and soft tissue infections: recognizing a broadening spectrum of disease*. Surg Infect 11(1): 49–57.
- Hidalgo E, Dominguez C. 2001. Mechanisms underlying chlorhexidine-induced cytotoxicity. Toxicol In Vitro 15(4): 271–6.
- Jagetia GC, Rajanikant GK. 2012. Acceleration of wound repair by curcumin in the excision wound of mice exposed to different doses of fractionated γ radiation. Int Wound J 9(1): 76–92.
- James GA, Swogger E, Wolcott R, et al. 2008. Biofilms in chronic wounds. Wound Repair Regen 16(1): 37–44.
- Johnson EN, Burns TC, Hayda RA, Hospenthal DR, Murray CK. 2007. Infectious complications of open type III tibial fractures among combat casualties. Clin Infect Dis 1545(4): 409–15.
- Jones V, Grey JE, Harding KG. 2006. Wound dressings. BMJ 332(7544): 777–780.Accessed May 11, 2015, at: http://www.oxforddictionaries.com/.
- Kalmes M, Blömeke B. 2012. Impact of eugenol and isoeugenol on AhR translocation, target gene expression, and proliferation in human HaCaT keratinocytes. J Toxicol Environ Health Part A 75(8-10): 478–91.
- Kamath JV, Rana AC, Roy CA. 2003. Pro-healing effect of Cinnamomum zeylanicum bark. Phytother Res 17(8): 970–2.
- Katti DS, Robinson KW, Ko FK, Laurencin CT. 2004. Bioresorbable nanofiber-based systems for wound healing and drug delivery: optimization of fabrication parameters. J Biomed Mater Res Part B Appl Biomater 70(2): 286–96.
- Knobloch K, Weigand H, Weis N, Schwarm HM, Vigenschow H. 1986. Action of Terpenoids on Energy Metabolism. Walter de Gruyter: Berlin, Germany.
- Liakos I, Rizzello L, Scurr DJ, Pompa PP, Bayer IS, Athanassiou A. 2014. All-natural composite wound dressing films of essential oils encapsulated in sodium alginate with antimicrobial properties. Int J Pharm 463(2): 137–45.
- Liou ML, Soo PC, Ling SR, Kuo HY, Tang CY, Chang KC. 2014. The sensor kinase BfmS mediates virulence in Acinetobacter baumannii. J Microbiol Immunol Infect 47(4): 275–81.
- Loehfelm TW, Luke NR, Campagnari AA. 2008. Identification and characterization of an Acinetobacter baumannii biofilm-associated protein. J Bacteriol 190: 1036–1044.
- Longo F, Vuotto C, Donelli G. 2014. Biofilm formation in Acinetobacter baumannii. New Microbiol 37(2): 119–27.
- Mackowiak PA. 1982. The normal microbial flora. N Engl J Med 307(2): 83–93.
- Martin JM, Zenilman JM, Lazarus GS. 2010. Molecular microbiology: new dimensions for cutaneous biology and wound healing. J Invest Dermatol 130(1): 38–48.
- Murray CK, Hsu JR, Solomkin JS, et al. 2008. Prevention and management of infections associated with combat-related extremity injuries. J Trauma 64: S239–51.
- Muthaiyan A, Biswas D, Crandall PG, Wilkinson BJ, Ricke SC. 2012. Application of orange essential oil as an antistaphylococcal agent in a dressing model. BMC Complement Altern Med 12(1): 1.
- Negi PS. 2012. Plant extracts for the control of bacterial growth: efficacy, stability and safety issues for food application. Int J Food Microbiol 156: 7–17.
- Ni Y, Yates KM, Tizard IR. 2004. In Aloes: The Genus Aloe, T Reynolds (ed). CRC Press LLC.: NewYork.
- Niu C, Clemmer KM, Bonomo RA, Rather PN. 2008. Isolation and characterization of an autoinducer synthase from Acinetobacter baumannii. J Bacteriol 190(9): 3386–3392.
- Oussalah M, Caillet S, Lacroix M. 2006. Mechanism of action of Spanish oregano, Chinese cinnamon, and savory essential oils against cell membranes and walls of Escherichia coli O157: H7 and Listeria monocytogenes. J Food Prot ® 69: 1046–1055.
- Peleg AY, Seifert H, Paterson DL. 2008. Acinetobacter baumannii: emergence of a successful pathogen. Clin Microbiol Rev 21(3): 538–82.
- Percival SL, Emanuel C, Cutting KF, Williams DW. 2012. Microbiology of the skin and the role of biofilms in infection. Int Wound J 9: 14–32.
- Price PB. 1938. The bacteriology of normal skin; a new quantitative test applied to a study of the bacterial flora and the disinfectant action of mechanical cleansing. J Infect Dis 11(1): 301–318.
10.1093/infdis/63.3.301 Google Scholar
- Rastogi VK, Wallace L, Smith LS. 2007. Disinfection of Acinetobacter baumannii-contaminated surfaces relevant to medical treatment facilities with ultraviolet C light. Mil Med 172: 1166–1169.
- Santucci SG, Gobara S, Santos CR, Fontana C, Levin AS. 2003. Infections in a burn intensive care unit: experience of seven years. J Hosp Infect 53: 6–13.
- Schröder JM, Harder J. 2006. Antimicrobial skin peptides and proteins. Cell Mol Life Sci 63(4): 469–86.
- Scott P, Deye G, Srinivasan A, Murray C, Moran K, Hulten E. 2007. An outbreak of multidrug-resistant Acinetobacter baumannii-calcoaceticus complex infection in the US military health care system associated with military operations in Iraq. Clin Infect Dis 44: 1577–1584.
- Seifert H, Dijkshoorn L, Gerner-Smidt P, Pelzer N, Tjernberg I, Vaneechoutte M. 1997. Distribution of Acinetobacter species on human skin: comparison of phenotypic and genotypic identification methods. J Clin Microbiol 35: 2819–2825.
- Selçuk CT, Durgun M, Özalp B, et al. 2012. Comparison of the antibacterial effect of silver sulfadiazine 1%, mupirocin 2%, Acticoat and octenidine dihydrochloride in a full-thickness rat burn model contaminated with multi drug resistant Acinetobacter baumannii. Burns 38(8): 1204–9.
- Sharma BR. 2007. Infection in patients with severe burns: causes and prevention thereof. Infect Dis Clin North Am 21(3): 745–59.
- Sikkema J, Weber FJ, Heipeiper HJ, Bont JAD. 1994. Cellular toxicity of lipophilic compounds:mechanisms, implications, and adaptations. Biocatalysis 10(1–4): 113–122.
- Thompson MG, Black CC, Pavlicek RL. 2014. Validation of a novel murine wound model of Acinetobacter baumannii infection. Antimicrob Agents Chemother 58(3): 1332–42.
- Thoroski J, Blank G, Biliaderis C. 1989. Eugenol induced inhibition of extracellular enzyme production by Bacillus subtilis. J Food Prot 52: 399–403.
- Upadhyay A, Upadhyaya I, Kollanoor-Johny A, Venkitanarayanan K. 2013. Antibiofilm effect of plant derived antimicrobials on Listeria monocytogenes. Food Microbiol31; 36(1): 79–89.
- Ultee A, Bennik MHJ, Moezelaar R. 2002. The phenolic hydroxyl group of carvacrol is essential for action against the food-borne pathogen Bacillus cereus. Appl Environ Microbiol 68(4): 1561–1568.
- Uygur F, Öncül O, Evinç R, Diktas H, Acar A, Ülkür E. 2009. Effects of three different topical antibacterial dressings on Acinetobacter baumannii-contaminated full- thickness Burns in rats. Burns 35: 270–273.
- Werthen M, Henriksson L, Jensen PØ, Sternberg C, Givskov M, Bjarnsholt T. 2010. An in vitro model of bacterial infections in wounds and other soft tissues. Apmis 118: 156–164.
- Woollard AC, Tatham KC, Barker S. 2007. The influence of essential oils on the process of wound healing: a review of the current evidence. J Wound Care 16(6): 255–7.
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