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Hydrogels Containing Antibiofilm and Antimicrobial Agents Beneficial for Biofilm-Associated Wound Infection: Formulation Characterizations and In vitro Study

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Abstract

Bacterial biofilm which adheres onto wound surface is shown to be impervious to antibiotics and this in turn delays wound healing. Previous studies showed that antibiofilm agents such as xylitol and ethylenediaminetetraacetic acid (EDTA) prevent bacterial adherence onto surfaces. Formulation of a wound dressing containing antibiofilm agents may be a plausible strategy in breaking the biofilm on wound surfaces and at the same time increase the efficacy of the antibiotic. The purpose of this study was to develop hydrogel formulations containing antibiofilm agents along with antibiotic (gentamicin) for bacterial biofilm-associated wound infection. Sodium carboxymethyl cellulose (NaCMC) hydrogels loaded with antibiofilm agents and antibiotic were prepared. The hydrogels were characterized for their physical properties, rheology, Fourier transform infrared spectroscopy (FTIR), drug content uniformity, differential scanning calorimetry (DSC) and in vitro drug release study. The antibiofilm (Crystal Violet staining and XTT assay) and antibacterial performances of the hydrogels against Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, and Escherichia coli were assessed in vitro. The formulated hydrogels showed adequate release of both antibiofilm agents (xylitol and EDTA). Both antimicrobial and antibiofilm tests showed promising results and demonstrated that the combination of xylitol, EDTA, and gentamicin had an additive effect against both Gram-positive and Gram-negative bacteria. In summary, NaCMC (sodium carboxymethyl cellulose) hydrogels containing the combination of antimicrobial and antibiofilm agents were successfully developed and this can be a new strategy in combating biofilm in wound infection which in turn accelerate wound healing.

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References

  1. Kirketerp-Møller K, Zulkowski K, James G. Chronic wound colonization, infection, and biofilms. In: Biofilm infections. New York: Springer; 2011. p. 11–24.

    Chapter  Google Scholar 

  2. Wolcott R, Cutting KF, Dowd S. Surgical site infections: biofilms, dehiscence and delayed healing. Wounds UK. 2008;4(4):108–3.

    Google Scholar 

  3. Ryan TJ. Infection following soft tissue injury: its role in wound healing. Curr Opin Infect Dis. 2007;20(2):124–8. https://doi.org/10.1097/QCO.0b013e32801a3e7c.

    Article  PubMed  Google Scholar 

  4. Ammons MC. Anti-biofilm strategies and the need for innovations in wound care. Recent patents on anti-infective drug discovery. 2010;5(1):10–7. https://doi.org/10.2174/157489110790112581.

    Article  CAS  PubMed  Google Scholar 

  5. Gawande PV, Yakandawala N, Vetri KL, Madhyashtha S. In vitro antimicrobial and antibiofilm activity of DispersinB-Tricolsan wound gel against chronic wound-associated bacteria. Open Antimicrob Agents J. 2011;3(1):12–6. https://doi.org/10.2174/1876518101103010012.

    Article  CAS  Google Scholar 

  6. Masako K, Hideyuki I, Shigeyuki O, Zenro IA. Novel method to control the balance of skin microflora: part 1. Attack on biofilm of Staphylococcus aureus without antibiotics. J Dermatol Sci. 2005;38(3):197–205. https://doi.org/10.1016/j.jdermsci.2005.01.006.

    Article  CAS  PubMed  Google Scholar 

  7. Chang Y, Gu W, McLandsborough L. Low concentration of ethylenediaminetetraacetic acid (EDTA) affects biofilm formation of Listeria monocytogenes by inhibiting its initial adherence. Food Microbiol. 2012;29(1):10–7. https://doi.org/10.1016/j.fm.2011.07.009.

    Article  CAS  PubMed  Google Scholar 

  8. Finnegan S, Percival SLEDTA. An antimicrobial and antibiofilm agent for use in wound care. Advances in wound care. 2015;4(7):415–21. https://doi.org/10.1089/wound.2014.0577.

    Article  PubMed Central  PubMed  Google Scholar 

  9. Percival SL, Cutting KF. Biofilms: possible strategies for suppression in chronic wounds. Nurs Stand. 2009;23(32):64–72.

    CAS  PubMed  Google Scholar 

  10. Lanigan RS, Yamarik TA. Final report on the safety assessment of EDTA, calcium disodium EDTA, diammonium EDTA, dipotassium EDTA, disodium EDTA, TEA-EDTA, tetrasodium EDTA, tripotassium EDTA, trisodium EDTA, HEDTA, and trisodium HEDTA. Int J Toxicol. 2002;21:95–142.

    Article  CAS  PubMed  Google Scholar 

  11. Riley P, Moore D, Ahmed F, Sharif MO, Worthington HV. Xylitol-containing products for preventing dental caries in children and adults. Cochrane Libr. 2015;

  12. Henry-Stanley MJ, Hess DJ, Wells CL. Aminoglycoside inhibition of Staphylococcus aureus biofilm formation is nutrient dependent. J Med Microbiol. 2014;63(6):861–9. https://doi.org/10.1099/jmm.0.068130-0.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  13. Vaneau M, Chaby G, Guillot B, Martel P, Senet P, Téot L, et al. Consensus panel recommendations for chronic and acute wound dressings. Arch Dermatol. 2007;143(10):1291–4. https://doi.org/10.1001/archderm.143.10.1291.

    Article  PubMed  Google Scholar 

  14. Ng SF, Leow HL. Development of biofilm-targeted antimicrobial wound dressing for the treatment of chronic wound infections. Drug Dev Ind Pharm. 2015;41(11):1902–9. https://doi.org/10.3109/03639045.2015.1019888.

    Article  CAS  PubMed  Google Scholar 

  15. Watts JL et al. Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals; approved standard. 3rd ed. Wayne: Clinical and Laboratory Standards Institute; 2008. p. M31–A3.

  16. Christensen GD, Simpson WA, Younger JJ, Baddour LM, Barrett FF, Melton DM, et al. Adherence of coagulase-negative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci to medical devices. J Clin Microbiol. 1985;22(6):996–1006.

    CAS  PubMed Central  PubMed  Google Scholar 

  17. Wei GX, Campagna AN, Bobek LA. Effect of MUC7 peptides on the growth of bacteria and on Streptococcus mutans biofilm. J Antimicrob Chemother. 2006;57(6):1100–9. https://doi.org/10.1093/jac/dkl120.

    Article  CAS  PubMed  Google Scholar 

  18. Muscat D, Adhikari B, Adhikari R, Chaudhary DS. Comparative study of film forming behaviour of low and high amylose starches using glycerol and xylitol as plasticizers. J Food Eng. 2012;109(2):189–201. https://doi.org/10.1016/j.jfoodeng.2011.10.019.

    Article  CAS  Google Scholar 

  19. Matthews KH, Stevens HN, Auffret AD, Humphrey MJ, Eccleston GM. Lyophilised wafers as a drug delivery system for wound healing containing methylcellulose as a viscosity modifier. Int J Pharm. 2005;289(1):51–62. https://doi.org/10.1016/j.ijpharm.2004.10.022.

    Article  CAS  PubMed  Google Scholar 

  20. Stout EI, McKessor A. Glycerin-based hydrogel for infection control. Advances in wound care. 2012;1(1):48–51. https://doi.org/10.1089/wound.2011.0288.

    Article  PubMed Central  PubMed  Google Scholar 

  21. Navarra MA, Dal Bosco C, Serra Moreno J, Vitucci FM, Paolone A, Panero S. Synthesis and characterization of cellulose-based hydrogels to be used as gel electrolytes. Membranes. 2015;5(4):810–23. https://doi.org/10.3390/membranes5040810.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  22. Thürmer MB, Diehl CE, Brum FJ, Santos LA. Preparation and characterization of hydrogels with potential for use as biomaterials. Mater Res. 2014;17(suppl 1):109–13. https://doi.org/10.1590/1516-1439.223613.

    Article  Google Scholar 

  23. Qu B, Li JR, Xiao HN, He BH, Qian LY. Preparation of sodium carboxymethylcellulose/poly (methyl acrylate) IPN hydrogels and their application for adsorption. J Appl Polym Sci. 2014;131(22). https://doi.org/10.1002/app.41101

  24. Li W, Sun B, Study WP. On hydrogen bonds of carboxymethyl cellulose sodium film with two-dimensional correlation infrared spectroscopy. Carbohydr Polym. 2009;78(3):454–61. https://doi.org/10.1016/j.carbpol.2009.05.002.

    Article  CAS  Google Scholar 

  25. Siegel RA, Langer R. Mechanistic studies of macromolecular drug release from macroporous polymers. II. Models for the slow kinetics of drug release. J Control Release. 1990;14(2):153–67. https://doi.org/10.1016/0168-3659(90)90152-J.

    Article  CAS  Google Scholar 

  26. Root JL, McIntyre OR, Jacobs NJ, Daghlian CP. Inhibitory effect of disodium EDTA upon the growth of Staphylococcus epidermidis in vitro: relation to infection prophylaxis of Hickman catheters. Antimicrob Agents Chemother. 1988;32(11):1627–31. https://doi.org/10.1128/AAC.32.11.1627.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  27. Tapiainen T, Sormunen R, Kaijalainen T, Kontiokari T, Ikäheimo I, Uhari M. Ultrastructure of Streptococcus pneumoniae after exposure to xylitol. J Antimicrob Chemother. 2004;54(1):225–8. https://doi.org/10.1093/jac/dkh302.

    Article  CAS  PubMed  Google Scholar 

  28. Hassan A, Usman J, Kaleem F, Omair M, Khalid A, Iqbal M. Evaluation of different detection methods of biofilm formation in the clinical isolates. Braz J Infect Dis. 2011;15(4):305–11. https://doi.org/10.1016/S1413-8670(11)70197-0.

    Article  PubMed  Google Scholar 

  29. Mathur T, Singhal S, Khan S, Upadhyay DJ, Fatma T, Rattan A. Detection of biofilm formation among the clinical isolates of staphylococci: an evaluation of three different screening methods. Indian J Med Microbiol. 2006;24(1):25–9. https://doi.org/10.4103/0255-0857.19890.

    Article  CAS  PubMed  Google Scholar 

  30. Dhale RP, Ghorpade MV, Dharmadhikari CA. Comparison of various methods used to detect biofilm production of Candida species. J Clin Diagnostic Res: JCDR. 2014;8(11):DC18–c20. https://doi.org/10.7860/JCDR/2014/10445.5147.

    Google Scholar 

  31. Darouiche RO, Mansouri MD, Gawande PV, Madhyastha S. Antimicrobial and antibiofilm efficacy of triclosan and DispersinB® combination. J Antimicrob Chemother. 2009;64(1):88–93. https://doi.org/10.1093/jac/dkp158.

    Article  CAS  PubMed  Google Scholar 

  32. Rogers SA, Huigens RW, Cavanagh J, Melander C. Synergistic effects between conventional antibiotics and 2-aminoimidazole-derived antibiofilm agents. Antimicrob Agents Chemother. 2010;54(5):2112–8. https://doi.org/10.1128/AAC.01418-09.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  33. Donelli G, Francolini I, Romoli D, Guaglianone E, Piozzi A, Ragunath C, et al. Synergistic activity of dispersin B and cefamandole nafate in inhibition of staphylococcal biofilm growth on polyurethanes. Antimicrob Agents Chemother. 2007;51(8):2733–40. https://doi.org/10.1128/AAC.01249-06.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  34. Harrison JJ, Turner RJ, Joo DA, Stan MA, Chan CS, Allan ND, et al. Copper and quaternary ammonium cations exert synergistic bactericidal and antibiofilm activity against Pseudomonas aeruginosa. Antimicrob Agents Chemother. 2008;52(8):2870–81. https://doi.org/10.1128/AAC.00203-08.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  35. Mu H, Guo F, Niu H, Liu Q, Wang S, Duan J. Chitosan improves anti-biofilm efficacy of gentamicin through facilitating antibiotic penetration. Int J Mol Sci. 2014;15(12):22296–308. https://doi.org/10.3390/ijms151222296.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  36. Chauhan A, Lebeaux D, Ghigo JM, Beloin C. Full and broad-spectrum in vivo eradication of catheter-associated biofilms using gentamicin-EDTA antibiotic lock therapy. Antimicrob Agents Chemother. 2012;56(12):6310–8. https://doi.org/10.1128/AAC.01606-12.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  37. Donlan RM, Costerton JW. Biofilms: survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev. 2002;15(2):167–93. https://doi.org/10.1128/CMR.15.2.167-193.2002.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  38. Cai X, Luan Y, Dong Q, Shao W, Li Z, Zhao Z. Sustained release of 5-fluorouracil by incorporation into sodium carboxymethylcellulose sub-micron fibers. Int J Pharm. 2011;419(1):240–6. https://doi.org/10.1016/j.ijpharm.2011.07.008.

    Article  CAS  PubMed  Google Scholar 

  39. Behan RJ. Treatment of an open infected wound. Ann Surg. 1921;73(6):701–11. https://doi.org/10.1097/00000658-192106000-00003.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  40. Care RW. A study of biofilm-based wound management in subjects with critical limb ischaemia. J Wound Care. 2008;17(4):145.

    Article  Google Scholar 

  41. Warriner R, Burrell R. Infection and the chronic wound: a focus on silver. Adv Skin Wound Care. 2005;18(8):2–12. https://doi.org/10.1097/00129334-200510001-00001.

    Article  PubMed  Google Scholar 

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Acknowledgements

The authors would like to thank the Ministry of Science, Technology and Innovation Malaysia, MOSTI (Grant No. 02-01-02-SF1228) and the Faculty of Pharmacy, UKM, for their research facility support.

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Correspondence to Shiow-Fern Ng.

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Anjum, A., Sim, CH. & Ng, SF. Hydrogels Containing Antibiofilm and Antimicrobial Agents Beneficial for Biofilm-Associated Wound Infection: Formulation Characterizations and In vitro Study. AAPS PharmSciTech 19, 1219–1230 (2018). https://doi.org/10.1208/s12249-017-0937-4

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