• UTKRISHTA L RAJ Department of Biotechnology, Centre for Emerging Diseases, Jaypee Institute of Information Technology, A-10, Sector-62, Noida, Uttar Pradesh, India.
  • MEGHA GAUTAM Department of Biotechnology, Centre for Emerging Diseases, Jaypee Institute of Information Technology, A-10, Sector-62, Noida,Uttar Pradesh, India.
  • REEMA GABRANI Department of Biotechnology, Centre for Emerging Diseases, Jaypee Institute of Information Technology, A-10, Sector-62, Noida,Uttar Pradesh, India.


Objective: Trans-cinnamaldehyde (TC) has shown antimicrobial activity against various microorganisms, but its direct use has some disadvantages such as skin irritation, low bioavailability, and low solubility. The objective of the present work was to develop the oil-in-water nanoemulsions (NEs) of TC to enhance its antimicrobial activity against Escherichia coli.

Methods: The NEs of TC were prepared using triton x-100 and isopropyl alcohol as surfactant and cosurfactant. The developed NE was studied for size, zeta potential, and stability. NEs were evaluated for antimicrobial and antibiofilm activity against E. coli as indicator organism. NEs possible mode of action on E. coli was assessed by scanning electron microscopy (SEM).

Results: Stable NEs of TC exhibited a particle size of 210 nm and were able to inhibit the growth of planktonic as well as biofilm cultures of E. coli at 67 μg/ml. The ruthenium red staining indicated the inhibition of glycoprotein layer formation in extracellular matrix after treating with NE. TC-NE exhibited substantial decrease in E. coli growth as well as its viability at its minimum inhibitory concentration as determined by 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The mode of action of cinnamaldehyde through β-galactosidase assay on E. coli ML-35p strain indicated that it altered the bacterial cell membrane permeability. SEM results showed the presence of holes on the cell wall of the E. coli in the presence of TC-NE.

Conclusions: TC-NEs exhibited enhanced antimicrobial activity and were effective against E. coli biofilm. They also exhibited microbicidal activity and altered E. coli membrane permeability.

Keywords: Growth curve, MTT, Membrane permeability, Ruthenium red, SEM

Author Biography

UTKRISHTA L RAJ, Department of Biotechnology, Centre for Emerging Diseases, Jaypee Institute of Information Technology, A-10, Sector-62, Noida, Uttar Pradesh, India.

Associate Professor

Department of Biotechnology


1. G,Croxen MA, Finlay BB. Molecular mechanisms of Escherichia coli pathogenicity. Nat Rev Microbiol 2010;8:26-38.
2. Ito A, Taniuchi A, May T, Kawata K, Okabe S. Increased antibiotic resistance of Escherichia coli in mature biofilms. Appl Environ Microbiol 2009;75:4093-100.
3. Sabir N, Ikram A, Zaman G, Satti L, Gardezi A, Ahmed A, et al. Bacterial biofilm-based catheter-associated urinary tract infections: Causative pathogens and antibiotic resistance. Am J Infect Control 2017;45:1101-5.
4. Chandra H, Bishnoi P, Yadav A, Patni B, Mishra AP, Nautiyal AR, et al. Antimicrobial resistance and the alternative resources with special emphasis on plant-based antimicrobials-A review. Plants (Basel) 2017;6:E16.
5. Shah B, Davidson PM, Zhong Q. Nanocapsular dispersion of thymol for enhanced dispersibility and increased antimicrobial effectiveness against Escherichia coli O157:H7 and listeria monocytogenes in model food systems. Appl Environ Microbiol 2012;78:8448-53.
6. Balijepalli MK, Ayuba SB, Raghavendra S, Mallikarjuna RP. Cinnamomum genus: A review on its biological activities. Int J Pharm Pharm Sci 2017;9:1-11.
7. Nabavi SF, Di Lorenzo A, Izadi M, Sobarzo-Sánchez E, Daglia M, Nabavi SM, et al. Antibacterial effects of cinnamon: From farm to food, cosmetic and pharmaceutical industries. Nutrients 2015;7:7729-48.
8. Shan B, Cai YZ, Brooks JD, Corke H. Antibacterial properties and major bioactive components of cinnamon stick (Cinnamomum burmannii): Activity against foodborne pathogenic bacteria. J Agric Food Chem 2007;55:5484-90.
9. Sheen S, Huang CY, Ramos R, Chien SY, Scullen OJ, Sommers C, et al. Lethality prediction for Escherichia coli O157:H7 and uropathogenic E. coli in ground chicken treated with high pressure processing and trans-cinnamaldehyde. J Food Sci 2018;83:740-9.
10. Grullon J, Mack JP, Rojtman A. Using essential oils to combat the threat of multi-drug resistance bacteria P. aeruginosa. Int J Pharm Pharm Sci 2016;8:180-3.
11. Jo YJ, Chun JY, Kwon YJ, Min SG, Hong GP, Choi MJ. Physical and antimicrobial properties of trans-cinnamaldehyde nanoemulsions in water melon juice. LWT Food Sci Technol 2015;60:444-51.
12. Álvarez-Paino M, Muñoz-Bonilla A, Fernández-García M. Antimicrobial polymers in the nano-world. Nanomaterials (Basel) 2017;7:E48.
13. Demetzos C. Biophysics and thermodynamics: The scientific building blocks of bio-inspired drug delivery nano systems. AAPS PharmSciTech 2015;16:491-5.
14. Donsì F, Annunziata M, Vincensi M, Ferrari G. Design of nanoemulsion-based delivery systems of natural antimicrobials: Effect of the emulsifier. J Biotechnol 2012;159:342-50.
15. CLSI. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically; Approved Standard. 10th ed. Wayne PA: Clinical and Laboratory Standards Institute, CLSI Document; 2015. p. M07-A10.
16. Quave CL, Plano LR, Pantuso T, Bennett BC. Effects of extracts from Italian medicinal plants on planktonic growth, biofilm formation and adherence of methicillin-resistant Staphylococcus aureus. J Ethnopharmacol 2008;118:418-28.
17. Sharma G, Sharma S, Sharma P, Chandola D, Dang S, Gupta S, et al. Escherichia coli biofilm: Development and therapeutic strategies. J Appl Microbiol 2016;121:309-19.
18. Wang H, Cheng H, Wang F, Wei D, Wang X. An improved 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction assay for evaluating the viability of Escherichia coli cells. J Microbiol Methods 2010;82:330-3.
19. Prouty AM, Schwesinger WH, Gunn JS. Biofilm formation and interaction with the surfaces of gallstones by Salmonella spp. Infect Immun 2002;70:2640-9.
20. Donsì F, Annunziata M, Sessa M, Ferrari G. Nanoencapsulation of essential oils enhance their antimicrobial activity in foods. LWT Food Sci Technol 2011;44:1908-14.
21. Wang S, Su R, Nie S, Sun M, Zhang J, Wu D, et al. Application of nanotechnology in improving bioavailability and bioactivity of diet-derived phytochemicals. J Nutr Biochem 2014;25:363-76.
22. Anwer MK, Jamil S, Ibnouf EO, Shakeel F. Enhanced antibacterial effects of clove essential oil by nanoemulsion. J Oleo Sci 2014;63:347-54.
23. Moghimi R, Ghaderi L, Rafati H, Aliahmadi A, McClements DJ. Superior antibacterial activity of nanoemulsion of thymus daenensis essential oil against E. Coli. Food Chem 2016;194:410-5.
24. Quatrin PM, Verdi CM, de Souza ME, de Godoi SN, Klein B, Gundel A, et al. Antimicrobial and antibiofilm activities of nanoemulsions containing Eucalyptus globulus oil against Pseudomonas aeruginosa and Candida spp. Microb Pathog 2017;112:230-42.
25. Sharma G, Dang S, Gupta S, Gabrani R. Identification and molecular characterization of bacteria having antimicrobial and antibiofilm activity. Int J Pharm Pharm Sci 2016;8:111-4.
26. Song Z, Sun H, Yang Y, Jing H, Yang L, Tong Y, et al. Enhanced efficacy and anti-biofilm activity of novel nanoemulsions against skin burn wound multi-drug resistant MRSA infections. Nanomedicine 2016;12:1543-55.
27. Li YF, Sun HW, Gao R, Liu KY, Zhang HQ, Fu QH, et al. Inhibited biofilm formation and improved antibacterial activity of a novel nanoemulsion against cariogenic Streptococcus mutans in vitro and in vivo. Int J Nanomedicine 2015;10:447-62.
28. Jerobin J, Makwana P, Suresh Kumar RS, Sundaramoorthy R, Mukherjee A, Chandrasekaran N, et al. Antibacterial activity of neem nanoemulsion and its toxicity assessment on human lymphocytes in vitro. Int J Nanomedicine 2015;10 Suppl 1:77-86.
13 Views | 19 Downloads
How to Cite
Original Article(s)