Research Article

Nano-ciprofloxacin/meropenem exhibit bactericidal activity against Gram-negative bacteria and rescue septic rat model

https://orcid.org/0000-0002-8105-4531

Assiut International Center of Nanomedicine, Al-Rajhy Liver Hospital, Assiut University, Assiut, 71515, Egypt

Search for more papers by this author

Reham A Ibrahem

https://orcid.org/0000-0002-7050-5755

Department of Microbiology & Immunology, Faculty of Pharmacy, Minia University, Minia, 61511, Egypt

Search for more papers by this author

Rehab M Abd El-Baky

https://orcid.org/0000-0001-6094-5653

Department of Microbiology & Immunology, Faculty of Pharmacy, Minia University, Minia, 61511, Egypt

Department of Microbiology & Immunology, Faculty of Pharmacy, Deraya University, Minia, 61511, Egypt

Search for more papers by this author

Mahmoud Elsabahy

https://orcid.org/0000-0003-3341-1829

School of Biotechnology, Badr University in Cairo, Badr City, 11829, Egypt

Department of Chemistry, Texas A&M University, College Station, TX 77842, USA

Search for more papers by this author

Abeer MR Hussein

Pharmacology Department, Faculty of Medicine, Assiut University, Assiut, 71515, Egypt

Search for more papers by this author

Mohammed EM Tolba

Medical Parasitology Department, Faculty of Medicine, Assiut University, Assiut, 71515, Egypt

Search for more papers by this author

Sherine A Aly

*Author for correspondence: Tel.: +20 109 701 8789;

E-mail Address: s-aly71@aun.edu.eg

https://orcid.org/0000-0002-4486-325X

Department of Microbiology & Immunology, Faculty of Medicine, Assiut University, Assiut, 71515, Egypt

Search for more papers by this author

Published Online:7 Nov 2023https://doi.org/10.2217/nnm-2022-0314

View article

Abstract

Aim: We hypothesized that simultaneous administration of two antibiotics loaded into a nanopolymer matrix would augment their synergistic bactericidal interaction. Methods: Nanoplatforms of chitosan/Pluronic^® loaded with ciprofloxacin/meropenem (CS/Plu-Cip/Mer) were prepared by the ionic gelation method, using Plu at concentrations in the range 0.5–4% w/v. CS/Plu-Cip/Mer was evaluated for antibacterial synergistic activity in vitro and in vivo. Results: CS/Plu-Cip and CS/Plu-Mer with Plu concentrations of 3% w/v and 2% w/v, respectively, exhibited ∼80% encapsulation efficiency. The MICs of pathogens were fourfold to 16-fold lower for CS/Plu-Cip/Mer than for Cip/Mer. Synergy was evidenced for CS/Plu-Cip/Mer with a bactericidal effect (at 1× MIC and sub-MICs), and it significantly decreased bacterial load and rescued infected rats. Conclusion: This study illustrates the ability of CS/Plu nanopolymer to intensify synergy between antibiotics, thereby providing a promising potential to rejuvenate antibiotics considered ineffective against resistant pathogens.

Plain language summary

Antibiotics are used to treat bacterial infections. However, the more they are used, the less effective they become, because bacteria develop resistance to them. One strategy to overcome this is to treat bacterial infection with a combination of antibiotics that work well together. The antibiotics ciprofloxacin and meropenem are often given together to treat Pseudomonas aeruginosa, a bacterium which can cause sepsis, a type of blood poisoning. Another strategy to overcome antibiotic resistance is to load them into nanocarriers, which can change their properties. Nanocarrier-loaded antibiotics can reduce toxicity and increase effectiveness. This study investigated whether the effectiveness of this pair could be improved by loading them into nanoparticles. When these nanoparticles were given to rats with sepsis, they were significantly more effective than unloaded ciprofloxacin and meropenem combinations. These nanoparticles were also able to directly kill bacteria, rather than just prevent bacterial reproduction, as with the unloaded combination. This study demonstrates that nanocarrier loading can intensify the enhanced benefit of combined antibiotic treatments. This is a promising strategy to reuse antibiotics that have become ineffective at treating bacteria which have developed resistance.

Tweetable abstract

CS/Plu-Cip/Mer resulting in a bactericidal effect versus the Cip/Mer bacteriostatic effect. The CS/Plu-Cip/Mer resulted in statistically significant reductions in all measures tested in the septic rat model.

Graphical abstract

Nano-Cip/Mer enhanced the survival rate and decreased the bacterial load in a septic rat model.

Keywords:

Papers of special note have been highlighted as: • of interest; •• of considerable interest

References

1. Centers for Disease Control and Prevention. Antibiotic Resistance Threats in the United States, 2019. US Department of Health and Human Services, CDC, Atlanta, GA (2019).
Google Scholar
2. Tamma PD, Cosgrove SE, Maragakis LL. Combination therapy for treatment of infections with gram-negative bacteria. Clin. Microbiol. Rev. 25(3), 450–470 (2012).
Medline CASGoogle Scholar
3. Rees VE, Yadav R, Rogers KE et al. Meropenem combined with ciprofloxacin combats hypermutable Pseudomonas aeruginosa from respiratory infections of cystic fibrosis patients. Antimicrob. Agents Chemother. 62(11), e01150–18 (2018).
MedlineGoogle Scholar
4. Jiang Z, He X, Li J. Synergy effect of meropenem-based combinations against Acinetobacter baumannii: a systematic review and meta-analysis. Infect. Drug Resist. 11, 1083–1095 (2018).
Medline CASGoogle Scholar
5. Obuobi S, Julin K, Fredheim EGA, Johannessen M, Škalko-Basnet N. Liposomal delivery of antibiotic loaded nucleic acid nanogels with enhanced drug loading and synergistic anti-inflammatory activity against S. aureus intracellular infections. J. Control. Rel. 324, 620–632 (2020).
Medline CASGoogle Scholar
6. Raymond B. Five rules for resistance management in the antibiotic apocalypse, a road map for integrated microbial management. Evol. Appl. 12(6), 1079–1091 (2019).
MedlineGoogle Scholar
7. Yeh Y-C, Huang T-H, Yang S-C et al. Nano-based drug delivery or targeting to eradicate bacteria for infection mitigation: a review of recent advances. Front. Chem. 8, 286 (2020). • Discusses the advantages of antibacterial nanosystems for the treatment of infectious disorders.
Medline CASGoogle Scholar
8. Liu Z, Hadi MA, Aljuboory DS et al. High efficiency of Ag⁰ decorated Cu₂MoO₄ nanoparticles for heterogeneous photocatalytic activation, bactericidal system, and detection of glucose from blood sample. J. Photochem. Photobiol. B 236, 112571 (2022).
Medline CASGoogle Scholar
9. Bahadoran A, Baghbadorani NB, De Lile JR et al. Ag doped Sn₃O₄ nanostructure and immobilized on hyperbranched polypyrrole for visible light sensitized photocatalytic, antibacterial agent and microbial detection process. J. Photochem. Photobiol. B 228, 112393 (2022).
Medline CASGoogle Scholar
10. Syed A, Elgorban AM, Bahkali AH et al. Highly-impressive performances of novel NiCo₂O₄/Bi₂O₃/Ag₂ZrO₃ nanocomposites in photocatalysis system, removal pathway, toxicity estimation, and antibacterial activities. J. Taiwan Inst. Chem. Eng 149, DOI: 10.1016/j.jtice.2023.105004 (2023).
Google Scholar
11. Mu H, Tang J, Liu Q et al. Potent antibacterial nanoparticles against biofilm and intracellular bacteria. Sci. Rep. 6(1), 18877 (2016).
Medline CASGoogle Scholar
12. Mitchell MJ, Billingsley MM, Haley RM et al. Engineering precision nanoparticles for drug delivery. Nat. Rev. Drug Discov. 20(2), 101–124 (2021).
Medline CASGoogle Scholar
13. Abdelkader A, El-Mokhtar MA, Abdelkader O et al. Ultrahigh antibacterial efficacy of meropenem-loaded chitosan nanoparticles in a septic animal model. Carbohydr. Polym. 174, 1041–1050 (2017). •• Demonstrated the great efficacy of meropenem-loaded nanoparticles in improving survival rates and extraordinary bacterial clearance (i.e. ≥99%).
Medline CASGoogle Scholar
14. Aguilera-Correa JJ, Gisbert-Garzarán M, Mediero A et al. Arabic gum plus colistin coated moxifloxacin-loaded nanoparticles for the treatment of bone infection caused by Escherichia coli. Acta Biomater. 137, 218–237 (2022).
Medline CASGoogle Scholar
15. Hadiya S, Liu X, Abd El-Hammed W et al. Levofloxacin-loaded nanoparticles decrease emergence of fluoroquinolone resistance in Escherichia coli. Microb. Drug Resist. 24(8), 1098–1107 (2018). •• Demonstrated the effect of levofloxacin-loaded nanoparticles on decreasing the emergence of resistance in emerged mutants.
Medline CASGoogle Scholar
16. Hadiya S, Ibrahem RA, Abd El-Baky RM et al. Nanosized combined antimicrobial drugs decreased emergence of resistance in Escherichia coli: a future promise. Microb. Drug Resist. 28(10), 972–979 (2022). •• Demonstrated the effect of nanosized combined antibiotics on restricting the evolution of resistant mutants and decreasing the expression of the effux pump and porin.
Medline CASGoogle Scholar
17. Cheung RCF, Ng TB, Wong JH et al. Chitosan: an update on potential biomedical and pharmaceutical applications. Mar. Drugs 13(8), 5156–5186 (2015).
Medline CASGoogle Scholar
18. Yilmaz Atay H. Antibacterial activity of chitosan-based systems. In: Functional Chitosan. Jana SJana S (Eds). Springer, Singapore, 457–489 (2019).
Google Scholar
19. Sharmin S, Rahaman MM, Sarkar C et al. Nanoparticles as antimicrobial and antiviral agents: a literature-based perspective study. Heliyon 7(3), e06456 (2021).
Medline CASGoogle Scholar
20. Yu J, Qiu H, Yin S et al. Polymeric drug delivery system based on pluronics for cancer treatment. Molecules 26(12), 3610 (2021).
Medline CASGoogle Scholar
21. Hadiya S, Radwan R, Zakaria M et al. Nanoparticles integrating natural and synthetic polymers for in vivo insulin delivery. Pharm. Dev. Technol. 26(1), 30–40 (2020).
MedlineGoogle Scholar
22. Guan J, Cheng P, Huang S et al. Optimized preparation of levofloxacin-loaded chitosan nanoparticles by ionotropic gelation. Phys. Procedia 22, 163–169 (2011).
CASGoogle Scholar
23. Drago L, De Vecchi E, Nicola L et al. Activity of levofloxacin and ciprofloxacin in combination with cefepime, ceftazidime, imipenem, piperacillin-tazobactam and amikacin against different Pseudomonas aeruginosa phenotypes and Acinetobacter spp. Chemotherapy 50(4), 202–210 (2004). • Demonstrated that combining fluoroquinolones with β-lactams or amikacin increased action against P. aeruginosa and Acinetobacter spp.
Medline CASGoogle Scholar
24. Soudeiha MA, Dahdouh EA, Azar E et al. In vitro evaluation of the colistin–carbapenem combination in clinical isolates of A. baumannii using the checkerboard, Etest, and time–kill curve techniques. Front. Cell. Infect. Microbiol. 7, 209 (2017).
MedlineGoogle Scholar
25. Farooqui A, Khan A, Borghetto I et al. Synergistic antimicrobial activity of Camellia sinensis and Juglans regia against multidrug-resistant bacteria. PLOS ONE 10(2), e0118431 (2015).
MedlineGoogle Scholar
26. Bajaksouzian S, Visalli MA, Jacobs MR et al. Activities of levofloxacin, ofloxacin, and ciprofloxacin, alone and in combination with amikacin, against Acinetobacters as determined by checkerboard and time–kill studies. Antimicrob. Agents Chemother. 41(5), 1073–1076 (1997).
Medline CASGoogle Scholar
27. Petersen PJ, Labthavikul P, Jones CH et al. In vitro antibacterial activities of tigecycline in combination with other antimicrobial agents determined by chequerboard and time–kill kinetic analysis. J. Antimicrob. Chemother. 57(3), 573–576 (2006).
Medline CASGoogle Scholar
28. Toledo PVM, Aranha Junior AA, Arend LN et al. Activity of antimicrobial combinations against KPC-2-producing Klebsiella pneumoniae in a rat model and time–kill assay. Antimicrob. Agents Chemother. 59(7), 4301–4304 (2015).
Medline CASGoogle Scholar
29. Cirioni O, Ghiselli R, Silvestri C et al. Efficacy of the combination of tachyplesin III and clarithromycin in rat models of Escherichia coli sepsis. Antimicrob. Agents Chemother. 52(12), 4351–4355 (2008).
Medline CASGoogle Scholar
30. Gnanadhas DP, Ben Thomas M, Elango M et al. Chitosan–dextran sulphate nanocapsule drug delivery system as an effective therapeutic against intraphagosomal pathogen Salmonella. J. Antimicrob. Chemother. 68(11), 2576–2586 (2013).
Medline CASGoogle Scholar
31. Cirioni O, Giacometti A, Ghiselli R et al. LL-37 protects rats against lethal sepsis caused by Gram-negative bacteria. Antimicrob. Agents Chemother. 50(5), 1672–1679 (2006).
Medline CASGoogle Scholar
32. Sanders ER. Aseptic laboratory techniques: plating methods. J. Vis. Exp. (63), e3064 (2012).
MedlineGoogle Scholar
33. Lee N-Y, Ko W-C, Hsueh P-R. Nanoparticles in the treatment of infections caused by multidrug-resistant organisms. Front. Pharmacol. 10, 1153 (2019).
Medline CASGoogle Scholar
34. Chen Y, Jihad A, Hussam F et al. A facile preparation method for efficiency a novel LaNiO₃/SrCeO₃ (pn type) heterojunction catalyst in photocatalytic activities, bactericidal assessment and dopamine detection. Surf. Interfaces 38, 102830 (2023).
CASGoogle Scholar
35. Long W, Hamza MU, Abdul-Fattah MN et al. Preparation, photocatalytic and antibacterial studies on novel doped ferrite nanoparticles: characterization and mechanism evaluation. Colloids Surf. A Physicochem. Eng. Asp. 650, 129468 (2022).
CASGoogle Scholar
36. Hosseinzadeh H, Atyabi F, Dinarvand R et al. Chitosan–Pluronic nanoparticles as oral delivery of anticancer gemcitabine: preparation and in vitro study. Int. J. Nanomed. 7, 1851 (2012).
Medline CASGoogle Scholar
37. Krupka TM, Solorio L, Wilson RE et al. Formulation and characterization of echogenic lipid–Pluronic nanobubbles. Mol. Pharm. 7(1), 49–59 (2010).
Medline CASGoogle Scholar
38. Feris K, Otto C, Tinker J et al. Electrostatic interactions affect nanoparticle-mediated toxicity to Gram-negative bacterium Pseudomonas aeruginosa PAO1. Langmuir 26(6), 4429–4436 (2010).
Medline CASGoogle Scholar
39. García-Couce J, Tomás M, Fuentes G et al. Chitosan/Pluronic F127 thermosensitive hydrogel as an injectable dexamethasone delivery carrier. Gels 8(1), 44 (2022).
Medline CASGoogle Scholar
40. Prasanna A, Venkatasubbu GD. Sustained release of amoxicillin from hydroxyapatite nanocomposite for bone infections. Prog. Biomater. 7(4), 289–296 (2018).
Medline CASGoogle Scholar
41. Song W, Woo H, Kim J et al. In vitro activity of β-lactams in combination with other antimicrobial agents against resistant strains of Pseudomonas aeruginosa. Int. J. Antimicrob. Agents 21(1), 8–12 (2003).
Medline CASGoogle Scholar
42. Mayer I, Nagy E. Investigation of the synergic effects of aminoglycoside–fluoroquinolone and third-generation cephalosporin combinations against clinical isolates of Pseudomonas spp. J. Antimicrob. Chemother. 43(5), 651–657 (1999).
Medline CASGoogle Scholar
43. Erdem I, Kaynar-Tascioglu J, Kaya B et al. The comparison of in the vitro effect of imipenem or meropenem combined with ciprofloxacin or levofloxacin against multidrug-resistant Pseudomonas aeruginosa strains. Int. J. Antimicrob. Agents 20(5), 384–386 (2002).
Medline CASGoogle Scholar
44. Pendland SL, Messick CR, Jung R. In vitro synergy testing of levofloxacin, ofloxacin, and ciprofloxacin in combination with aztreonam, ceftazidime, or piperacillin against Pseudomonas aeruginosa. Diagn. Microbiol. Infect. Dis. 42(1), 75–78 (2002).
Medline CASGoogle Scholar
45. Wei S, Yang Y, Tian W et al. Synergistic activity of fluoroquinolones combining with artesunate against multidrug-resistant Escherichia coli. Microb. Drug Resist. 26(1), 81–88 (2020).
Medline CASGoogle Scholar
46. Wei W-J, Yang H-F. Synergy against extensively drug-resistant Acinetobacter baumannii in vitro by two old antibiotics: colistin and chloramphenicol. Int. J. Antimicrob. Agents. 49(3), 321–326 (2017).
Medline CASGoogle Scholar
47. Aguilera-Correa J, Gisbert-Garzarán M, Mediero A et al. Arabic gum plus colistin coated moxifloxacin-loaded nanoparticles for the treatment of bone infection caused by Escherichia coli. Acta Biomater. 137, 218–237 (2022). •• Demonstrated that nanoparticles were deemed a promising treatment for bone infections.
Medline CASGoogle Scholar
48. Drlica K. The mutant selection window and antimicrobial resistance. J. Antimicrob. Chemother. 52(1), 11–17 (2003).
Medline CASGoogle Scholar
49. Gianvecchio C, Lozano NA, Henderson C et al. Variation in mutant prevention concentrations. Front. Microbiol. 10, 42 (2019). • The study found that when the antibiotic concentration was higher than mutation prevention concentration, mutant subpopulation evolution was reduced.
MedlineGoogle Scholar
50. Brennan-Krohn T, Pironti A, Kirby JE. Synergistic activity of colistin-containing combinations against colistin-resistant Enterobacteriaceae. Antimicrob. Agents Chemother. 62(10), e00873–00818 (2018).
Medline CASGoogle Scholar
51. Cuero R, Osuji G, Washington A. N-carboxymethylchitosan inhibition of aflatoxin production: role of zinc. Biotechnol. Lett. 13, 441–444 (1991).
CASGoogle Scholar
52. Frei A, Verderosa AD, Elliott AG et al. Metals to combat antimicrobial resistance. Nat. Rev. Chem. 7(3), 202–224 (2023).
Medline CASGoogle Scholar
53. Brown D. Antibiotic resistance breakers: can repurposed drugs fill the antibiotic discovery void? Nat. Rev. Drug Discov. 14(12), 821–832 (2015).
Medline CASGoogle Scholar

Vol. 18, No. 22

Supplemental Materials

Metrics

Downloaded 51 times

History

Received 11 December 2022

Accepted 14 September 2023

Published online 7 November 2023

Published in print September 2023

Information

Keywords

Supplementary data

To view the supplementary data that accompany this paper please visit the journal website at: www.futuremedicine.com/doi/suppl/10.2217/nnm-2022-0314

Author contributions

S Hadiya: performed investigation, data analysis and writing (original draft). S Aly: performed experimental protocol conceptualization, project administration, supervision, accomplished the resources, validation and editing the manuscript. M Elsabahy: provided supervision of nanoparticle procedures, resources and supervision. R Abd El-Baky: performed validation and editing. R Ibrahem: supervision and editing. A Hussein and M Tolba: provided supervision of in vivo procedure and supervision.

Acknowledgments

The authors gratefully acknowledge S Ali (Tumor Biology Research Program, Children’s Cancer Hospital, Cairo 57357, Egypt) for his assistance with data analysis.

Financial disclosure

This study was supported financially by the Science and Technology Development Fund (STDF), Egypt, grant no. 25913. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.

Competing interests disclosure

The authors have no competing interests or relevant affiliations with any organization or entity with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.

Writing disclosure

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all animal experimental investigations. This study was authorized by Assiut University Animal Ethical Committee (IRB no. 17300370).

PDF download