Bacterial Ghosts Carrying 5-Fluorouracil: A Novel Biological Carrier for Targeting Colorectal Cancer

Abstract

Bacterial ghosts (BGs) are non-deformed bacterial cell envelopes that possess undamaged external configurations for precise attachment to different cells of the human body. The Escherichia coli BGs were successfully produced using a modified sponge-like reduced protocol and characterized by SEM. Four different concentrations of 5-fluorouracil (5-FU) were used to study the impact on the “ghosts” cell wall. 5-FU was then loaded into the BGs and the loading capacity (LC %) and entrapment efficiency (EE %) were determined and were found to be 38.3 ± 0.8 and 76.6 ± 0.8, respectively. The in vitro release studies were conducted in dialysis bags over a time period of 16 days and the accumulative 5-FU released (%) was calculated. Overall, 69.2% of the ghost-associated 5-FU was released from the BGs and release from the E. coli ghosts is governed by non-Fickian diffusion. The Caco-2 cell line was used to investigate the cytotoxicity of 5-FU-loaded BGs.

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References

  1. 1.

    Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, et al. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer. 2015;136:E359–86.

    CAS  Article  Google Scholar 

  2. 2.

    Joel GH, Lee EL, Alfred G. The pharmacological basis of therapeutics. International edition, 10th edition, Mc Grow Hill 2001, 971.

  3. 3.

    Arias JL. Novel strategies to improve the anticancer action of 5-fluorouracil by using drug delivery systems. Molecules. 2008;13:2340–69.

    CAS  Article  Google Scholar 

  4. 4.

    Michor F, Iwasa Y, Lengauer C, Nowak MA. Dynamics of colorectal cancer. Semin Cancer Biol. 2005;15:484–93.

    CAS  Article  Google Scholar 

  5. 5.

    Paukner S, Kohl G, Lubitz W. Bacterial ghosts as novel advanced drug delivery systems: antiproliferative activity of loaded doxorubicin in human Caco-2 cells. J Control Release. 2004;94:63–74.

    CAS  Article  Google Scholar 

  6. 6.

    Kudela P, Koller VJ, Lubitz W. Bacterial ghosts (BGs)—advanced antigen and drug delivery system. Vaccine. 2010;28:5760–7.

    CAS  Article  Google Scholar 

  7. 7.

    Andrews JM. Determination of minimum inhibitory concentrations. J Antimicrob Chemother. 2001;48:5–16.

    CAS  Article  Google Scholar 

  8. 8.

    Wiegand I, Hilpert K, Hancock RE. Agar and broth dilution methods to determine the minimal inhibitory concentration (MIC) of antimicrobial substances. Nat Protoc. 2008;3:163–75.

    CAS  Article  Google Scholar 

  9. 9.

    Sekar V. A rapid screening procedure for the identification of recombinant bacterial clones. BioTechniques. 1987;5(1):11–13.

  10. 10.

    Dwyer DJ, Camacho DM, Kohanski MA, Callura JM. Antibiotic-induced bacterial cell death exhibits physiological and biochemical hallmarks of apoptosis. Mol Cell. 2012;46:561–72.

    CAS  Article  Google Scholar 

  11. 11.

    Youssof AM, Salem-Bekhit MM, Shakeel F, Alanazi FK, Haq N. Analysis of anti-neoplastic drug in bacterial ghost matrix, w/o/w double nanoemulsion and w/o nanoemulsion by a validated ‘green’liquid chromatographic method. Talanta. 2016;154:292–8.

    CAS  Article  Google Scholar 

  12. 12.

    Gupta P, Hung C, Perrier D. Quantitation of the release of doxorubicin from colloidal dosage forms using dynamic dialysis. J Pharm Sci. 1987;76:141–5.

    CAS  Article  Google Scholar 

  13. 13.

    Nasr M, Ghorab MK, Abdelazem A. In vitro and in vivo evaluation of cubosomes containing 5-fluorouracil for liver targeting. Acta Pharm Sin B. 2015;5:79–88.

    Article  Google Scholar 

  14. 14.

    Amara AA, Salem-Bekhit MM, Alanazi FK. Sponge-like: a new protocol for preparing bacterial ghosts. Sci World J. 2013;2013:E545741.

    Article  Google Scholar 

  15. 15.

    Amara AA, Salem-Bekhit MM, Alanazi FK. Preparation of bacterial ghosts for E. coli JM109 using sponge-like reduced protocol. Asian J Biol Sci. 2013;6:363–9.

    CAS  Article  Google Scholar 

  16. 16.

    Tomasz A, Borek E. The mechanism of an osmotic instability induced in E. coli K-12 by 5-fluorouracil. Biochemist. 1962;1:543–52.

    CAS  Article  Google Scholar 

  17. 17.

    Tomasz A, Borek E. An early phase in the bactericidal action of 5-fluorouracil on E. Coli k12 osmotic imbalance. Proc Natl Acad Sci U S A. 1959;45:929–32.

    CAS  Article  Google Scholar 

  18. 18.

    Tomasz A, Borek E. The mechanism of bacterial fragility produced by 5-fluorouracil: the accumulation of cell wall precursors. Proc Natl Acad Sci U S A. 1960;46:324–7.

    CAS  Article  Google Scholar 

  19. 19.

    Nounou MM, El-Khordagui LK, Khalafallah N. Release stability of 5-fluorouracil liposomal concentrates, gels and lyophilized powder. Acta Pol Pharm. 2005;62:381–91.

    CAS  PubMed  Google Scholar 

  20. 20.

    Kataoka K, Matsumoto T, Yokoyama M, Okano T, Sakurai Y, Fukushima S, et al. Doxorubicin-loaded poly (ethylene glycol)–poly (β-benzyl-l-aspartate) copolymer micelles: their pharmaceutical characteristics and biological significance. J Control Release. 2000;64:143–53.

    CAS  Article  Google Scholar 

  21. 21.

    Wirth M, Fuchs A, Wolf M, Ertl B, Gabor F. Lectin-mediated drug targeting: preparation, binding characteristics, and antiproliferative activity of wheat germ agglutinin conjugated doxorubicin on Caco-2 cells. Pharm Res. 1998;15:1031–7.

    CAS  Article  Google Scholar 

  22. 22.

    Rahul KS, Kadam V, Shendarkar G, Jadhav S, Bharkad V. Sustained release drug delivery system: review. Ind J Res Pharm Biotechnol. 2011;3:246–51.

    Google Scholar 

  23. 23.

    Ozturk SS, Palsson BO, Donohoe B, Dressman JB. Kinetics of release from enteric-coated tablets. Pharm Res. 1988;5:550–65.

    CAS  Article  Google Scholar 

  24. 24.

    Higuchi T. Mechanism of sustained-action medication. Theoretical analysis of rate of release of solid drugs dispersed in solid matrices. J Pharm Sci. 1963;52:1145–9.

    CAS  Article  Google Scholar 

  25. 25.

    Tokumasu F, Ostera GR, Amaratunga C, Fairhurst RM. Modifications in erythrocyte membrane zeta potential by Plasmodium falciparum infection. Exp Parasitol. 2012;131:245–51.

    CAS  Article  Google Scholar 

  26. 26.

    Arakha M, Saleem M, Mallick BC, Jha S. The effects of interfacial potential on antimicrobial propensity of ZnO nanoparticle. Sci Rep. 2015;5:9578–87.

    CAS  Article  Google Scholar 

  27. 27.

    Klose CS, Flach M, Möhle L, Rogell L, Hoyler T, Ebert K, et al. Differentiation of type 1 ILCs from a common progenitor to all helper-like innate lymphoid cell lineages. Cell. 2014;157:340–56.

    CAS  Article  Google Scholar 

  28. 28.

    Mauad T, Hajjar LA, Callegari GD, Da Silva LF, Schout D, Galas FR, et al. Lung pathology in fatal novel human influenza A (H1N1) infection. American journal of respiratory and critical care medicine. Am J Respir Crit Care Med. 2010;181:72–9.

    Article  Google Scholar 

  29. 29.

    Alanazi FK, Harisa GE-DI, Maqboul A, Abdel-Hamid M, Neau SH, Alsarra IA. Biochemically altered human erythrocytes as a carrier for targeted delivery of primaquine: an in vitro study. Arch Pharm Res. 2011;34:563–71.

    CAS  Article  Google Scholar 

  30. 30.

    Wyllie AH. Apoptosis: an overview. Br Med Bull. 1997;53:451–65.

    CAS  Article  Google Scholar 

  31. 31.

    Stein E, Inic-Kanada A, Belij S, Montanaro J, Bintner N, Schlacher S, et al. In vitro and in vivo uptake study of Escherichia coli Nissle 1917 bacterial ghosts: cell-based delivery system to target ocular surface diseases Escherichia coli Nissle 1917 bacterial ghosts. Invest Ophthalmol Vis Sci. 2013;54:6326–33.

    CAS  Article  Google Scholar 

Download references

Acknowledgements

This project was financially supported by King Saud University, Vice Deanship of Research Chairs, Kayyali Chair for Pharmaceutical industry, through the Grant No. MS-2018.

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Correspondence to Nazrul Haq.

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Youssof, A.M.E., Alanazi, F.K., Salem-Bekhit, M.M. et al. Bacterial Ghosts Carrying 5-Fluorouracil: A Novel Biological Carrier for Targeting Colorectal Cancer. AAPS PharmSciTech 20, 48 (2019). https://doi.org/10.1208/s12249-018-1249-z

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KEY WORDS

  • targeting
  • 5-fluorouracil
  • bacterial ghost
  • colorectal cancer
  • cytotoxicity