Infected burn wound healing using Hydroxy-propyl-methyl cellulose gel containing bacteriophages against Pseudomonas aeruginosa and Klebsiella pneumoniae
-
Meysam Adibi
,
Kazem Javanmardi
,
Noor Al-Huda Ali A.H. Saeed
,
Mohammad Ali Mobasher
,
Javad Jokar
,
Abdolmajid Ghasemian
,
Niloofar Rahimian
,
Ava Soltani Hekmat
Abstract
Background and Objectives: Pseudomonas aeruginosa (P. aeruginosa) and Klebsiella pneumoniae (K. pneumoniae) are the two leading bacterial strains involved in wound infections. These bacteria have developed broad resistance to antibiotics, which has complicated their eradication. Additionally, the formation of a polymicrobial infection poses additional problems. Among alternative or complementary options, bacteriophages, viruses that parasitize bacterial hosts, have been promising.
Materials and Methods: In this research work, bacteriophages' therapeutic effects against P. aeruginosa- and K. pneumoniae-infected burn wounds were studied. The infectious burn wound model was performed on Balb/C male mice, aged six weeks and weighing 25 ± 5 gr. The effects of the Hydroxy-propyl-methyl cellulose (HPMC) gel containing phage were investigated compared to gentamicin. All of these actions were performed in separate groups for each bacteria and mixed group of bacteria (to test multi-bacterial infections treating) and the result were compared.
Results: Phages appear to be effective in gel forms. Pathologic samples of different groups confirmed therapeutic results of phages. These results at the microscopic level indicated the recovery of the tissue and the removal of the infection.
Conclusion: The results of this study indicate that lytic phages are powerful biological tools for the treatment of bacterial infections in burn wounds, which can be considered as one of the alternatives for drug-resistant bacterial species and the high costs of antibiotics; though further animal and trial studies are needed. Meanwhile, the complications due to their widespread use in humans should be investigated in more details.
1. Taheri F, Hashemi A, Haghighi M, Dadashi M, Nasiri MJ, Khoshnood S, et al. Characterization of Staphylococcus aureeus isolated from burn wound; strong antibacterial activity of phage cocktail against vancomycin intermedlate-resistant Staphyl ocoous aureus. J Microbiol Biotechnol Food Sci 2022; 12(2): e5366.
2. Maitz J, Merlino J, Rizzo S, McKew G, Maitz P. Burn wound infections microbiome and novel approaches using therapeutic microorganisms in burn wound infection control. Adv Drug Deliv Rev 2023; 196: 114769.
3. Özlü Ö, Basaran A. Epidemiology and outcome of 1442 pediatric burn patients: A single-center experience. Ulus Travma Acil Cerrahi Derg 2022; 28: 57-61.
4. Omar A, Wright JB, Schultz G, Burrell R, Nadworny P. Microbial biofilms and chronic wounds. Microorganisms 2017; 5: 9.
5. Ruegsegger L, Xiao J, Naziripour A, Kanumuambidi T, Brown D, Williams F, et al. Multidrug-resistant gram-negative bacteria in burn patients. Antimicrob Agents Chemother 2022; 66(9): e0068822.
6. Buch PJ, Chai Y, Goluch ED. Treating polymicrobial infections in chronic diabetic wounds. Clin Microbiol Rev 2019; 32(2): e00091-18.
7. Krezalek MA, Alverdy JC. The influence of intestinal microbiome on wound healing and infection. Semin Colon Rectal Surg 2018; 29: 17-20.
8. Organization WH (2014). Antimicrobial resistance: global report on surveillance: World Health Organization. https://www.who.int/publications/i/item/9789241564748
9. Tang KWK, Millar BC, Moore JE. Antimicrobial resistance (AMR). Br J Biomed Sci 2023; 80: 11387.
10. Toprak E, Veres A, Michel J-B, Chait R, Hartl DL, Kishony R. Evolutionary paths to antibiotic resistance under dynamically sustained drug selection. Nat Genet 2011; 44: 101-105.
11. Kapoor G, Saigal S, Elongavan A. Action and resistance mechanisms of antibiotics: A guide for clinicians. J Anaesthesiol Clin Pharmacol 2017; 33: 300-305.
12. Simpkin VL, Renwick MJ, Kelly R, Mossialos E. Incentivising innovation in antibiotic drug discovery and development: progress, challenges and next steps. J Antibiot (Tokyo) 2017; 70: 1087-1096.
13. Golkar Z, Bagasra O, Pace DG. Bacteriophage therapy: a potential solution for the antibiotic resistance crisis. J Infect Dev Ctries 2014; 8: 129-136.
14. Kortright KE, Chan BK, Koff JL, Turner PE. Phage therapy: a renewed approach to combat antibiotic-resistant bacteria. Cell Host Microbe 2019; 25: 219-232.
15. Lorch A. Bacteriophages: An alternative to antibiotics? Aust Biotechnol 1999; 9: 265-269.
16. Pinto AM, Cerqueira MA, Bañobre-Lópes M, Pastrana LM, Sillankorva S. Bacteriophages for chronic wound treatment: From traditional to novel delivery systems. Viruses 2020; 12: 235.
17. Karn SL, Gangwar M, Kumar R, Bhartiya SK, Nath G. Phage therapy: a revolutionary shift in the management of bacterial infections, pioneering new horizons in clinical practice, and reimagining the arsenal against microbial pathogens. Front Med (Lausanne) 2023; 10: 1209782.
18. Rezaei Z, Elikaei A, Barzi SM, Shafiei M. Isolation, characterization, and antibacterial activity of lytic bacteriophage against methicillin-resistant Staphylococcus aureus causing bedsore and diabetic wounds. Iran J Microbiol 2022; 14: 712-720.
19. Lashtoo Aghaee B, Alikhani MY, van Leeuwen WB, Mojtahedi A, Kazemi S, Karami P. Conventional Treatment of burn wound infections versus phage therapy. Iran J Med Microbiol 2022; 16: 186-196.
20. Torabi LR, Naghavi NS, Doudi M, Monajemi R. Efficacious antibacterial potency of novel bacteriophages against ESBL-producing Klebsiella pneumoniae isolated from burn wound infections. Iran J Microbiol 2021; 13: 678-690.
21. Cerveny KE, DePaola A, Duckworth DH, Gulig PA. Phage therapy of local and systemic disease caused by Vibrio vulnificus in iron-dextran-treated mice. Infect Immun 2002; 70: 6251-6262.
22. Chang H-C, Chen C-R, Lin J-W, Shen G-H, Chang K-M, Tseng Y-H, et al. Isolation and characterization of novel giant Stenotrophomonas maltophilia phage phiSMA5. Appl Environ Microbiol 2005; 71: 1387-1393.
23. Jo SJ, Lee YM, Cho K, Park SY, Kwon H, Giri SS, et al. Standardization of the Agar plate method for bacteriophage production. Antibiotics 2025; 14: 2.
24. Peters DL, Harris G, Davis CM, Dennis JJ, Chen W. Bacteriophage isolation, purification, and characterization techniques against ubiquitous opportunistic pathogens. Curr Protoc 2022; 2(11): e594.
25. Skaradzińska A, Ochocka M, Śliwka P, Kuźmińska-Bajor M, Skaradziński G, Friese A, et al. Bacteriophage amplification–A comparison of selected methods. J Virol Methods 2020; 282: 113856.
26. Lapras B, Marchand C, Merienne C, Medina M, Kolenda C, Laurent F, et al. Rationalisation of the purification process for a phage active pharmaceutical ingredient. Eur J Pharm Biopharm 2024: 203: 114438.
27. Biswas B, Adhya S, Washart P, Paul B, Trostel AN, Powell B, et al. Bacteriophage therapy rescues mice bacteremic from a clinical isolate of vancomycin-resistant Enterococcus faecium. Infect Immun 2002; 70: 204-210.
28. Nikumbh KV, Sevankar SG, Patil MP. Formulation development, in vitro and in vivo evaluation of microemulsion-based gel loaded with ketoprofen. Drug Deliv 2015; 22: 509-515.
29. Jokar J, Abdulabbas HT, Javanmardi K, Mobasher MA, Jafari S, Ghasemian A, et al. Enhancement of bactericidal effects of bacteriophage and gentamicin combination regimen against Staphylococcus aureus and Pseudomonas aeruginosa strains in a mice diabetic wound model. Virus Genes 2024; 60: 80-96.
30. Björn C, Noppa L, Näslund Salomonsson E, Johansson A-L, Nilsson E, Mahlapuu M, et al. Efficacy and safety profile of the novel antimicrobial peptide PXL150 in a mouse model of infected burn wounds. Int J Antimicrob Agents 2015; 45: 519-524.
31. Shrum B, Anantha RV, Xu SX, Donnelly M, Haeryfar SM, McCormick JK, et al. A robust scoring system to evaluate sepsis severity in an animal model. BMC Res Notes 2014; 7: 233.
32. Brans TA, Dutrieux RP, Hoekstra MJ, Kreis RW, du Pont JS. Histopathological evaluation of scalds and contact burns in the pig model. Burns 1994; 20 Suppl 1: S48-S51.
33. Jokar J, Saleh RO, Rahimian N, Ghasemian A, Ghaznavi G, Radfar A, et al. Antibacterial effects of single phage and phage cocktail against multidrug-resistant Klebsiella pneumoniae isolated from diabetic foot ulcer. Virus Genes 2023; 59: 635-642.
34. Jamalludeen N, Shakir D. In vitro effect of newly bacteriophages isolates against Escherichia coli serogroups collected from the local hospital laboratory; their specific characterization and identification. J Microbiol Biotechnol Food Sci 2022; 11(6): e5478.
35. Gupta M, Naik AK, Singh SK. Bacteriological profile and antimicrobial resistance patterns of burn wound infections in a tertiary care hospital. Heliyon 2019; 5(12): e02956.
36. Litwin A, Rojek S, Gozdzik W, Duszynska W. Pseudomonas aeruginosa device associated–healthcare associated infections and its multidrug resistance at intensive care unit of University Hospital: polish, 8.5-year, prospective, single-centre study. BMC Infect Dis 2021; 21: 180.
37. Wyres KL, Lam MMC, Holt KE. Population genomics of Klebsiella pneumoniae. Nat Rev Microbiol 2020; 18: 344-359.
38. Azeredo J, Sutherland IW. The use of phages for the removal of infectious biofilms. Curr Pharm Biotechnol 2008; 9: 261-266.
39. Sharma S, Datta S, Chatterjee S, Dutta M, Samanta J, Vairale MG, et al. Isolation and characterization of a lytic bacteriophage against Pseudomonas aeruginosa. Sci Rep 2021; 11: 19393.
40. Li N, Zeng Y, Bao R, Zhu T, Tan D, Hu B. Isolation and characterization of novel phages targeting pathogenic Klebsiella pneumoniae. Front Cell Infect Microbiol 2021; 11: 792305.
41. Gibran NS, Boyce S, Greenhalgh DG. Cutaneous wound healing. J Burn Care Res 2007; 28: 577-579.
42. Weber-Dabrowska B, Mulczyk M, Górski A. Bacteriophage therapy for infections in cancer patients. Clin Appl Immun Rev 2001; 1: 131-134.
43. Kumari S, Harjai K, Chhibber S. Evidence to support the therapeutic potential of bacteriophage Kpn5 in burn wound infection caused by Klebsiella pneumoniae in BALB/c mice. J Microbiol Biotechnol 2010; 20: 935-941.
44. Finlay‐Jones JJ, Davies KV, Sturm LP, Kenny PA, Hart PH. Inflammatory processes in a murine model of intra‐abdominal abscess formation. J Leukoc Biol 1999; 66: 583-587.
45. Guerra MES, Destro G, Vieira B, Lima AS, Ferraz LFC, Hakansson AP, et al. Klebsiella pneumoniae biofilms and their role in disease pathogenesis. Front Cell Infect Microbiol 2022; 12: 877995.
46. Soothill J. Use of bacteriophages in the treatment of Pseudomonas aeruginosa infections. Expert Rev Anti Infect Ther 2013; 11: 909-915.
47. Kumari S, Harjai K, Chhibber S. Bacteriophage versus antimicrobial agents for the treatment of murine burn wound infection caused by Klebsiella pneumoniae B5055. J Med Microbiol 2011; 60: 205-210.
48. Punjataewakupt A, Napavichayanun S, Aramwit P. The downside of antimicrobial agents for wound healing. Eur J Clin Microbiol Infect Dis 2019; 38: 39-54.
49. Shevchenko OV, Kharina AV, Korniienko NO, Budzanivska IH, Andriichuk OM, Pozhylov IM, et al. Phage therapy in traumatology: A review on perspectives for treating acute wounds and Post-Surgical complications. Mikrobiol Zh 2024; 86: 117-136.
2. Maitz J, Merlino J, Rizzo S, McKew G, Maitz P. Burn wound infections microbiome and novel approaches using therapeutic microorganisms in burn wound infection control. Adv Drug Deliv Rev 2023; 196: 114769.
3. Özlü Ö, Basaran A. Epidemiology and outcome of 1442 pediatric burn patients: A single-center experience. Ulus Travma Acil Cerrahi Derg 2022; 28: 57-61.
4. Omar A, Wright JB, Schultz G, Burrell R, Nadworny P. Microbial biofilms and chronic wounds. Microorganisms 2017; 5: 9.
5. Ruegsegger L, Xiao J, Naziripour A, Kanumuambidi T, Brown D, Williams F, et al. Multidrug-resistant gram-negative bacteria in burn patients. Antimicrob Agents Chemother 2022; 66(9): e0068822.
6. Buch PJ, Chai Y, Goluch ED. Treating polymicrobial infections in chronic diabetic wounds. Clin Microbiol Rev 2019; 32(2): e00091-18.
7. Krezalek MA, Alverdy JC. The influence of intestinal microbiome on wound healing and infection. Semin Colon Rectal Surg 2018; 29: 17-20.
8. Organization WH (2014). Antimicrobial resistance: global report on surveillance: World Health Organization. https://www.who.int/publications/i/item/9789241564748
9. Tang KWK, Millar BC, Moore JE. Antimicrobial resistance (AMR). Br J Biomed Sci 2023; 80: 11387.
10. Toprak E, Veres A, Michel J-B, Chait R, Hartl DL, Kishony R. Evolutionary paths to antibiotic resistance under dynamically sustained drug selection. Nat Genet 2011; 44: 101-105.
11. Kapoor G, Saigal S, Elongavan A. Action and resistance mechanisms of antibiotics: A guide for clinicians. J Anaesthesiol Clin Pharmacol 2017; 33: 300-305.
12. Simpkin VL, Renwick MJ, Kelly R, Mossialos E. Incentivising innovation in antibiotic drug discovery and development: progress, challenges and next steps. J Antibiot (Tokyo) 2017; 70: 1087-1096.
13. Golkar Z, Bagasra O, Pace DG. Bacteriophage therapy: a potential solution for the antibiotic resistance crisis. J Infect Dev Ctries 2014; 8: 129-136.
14. Kortright KE, Chan BK, Koff JL, Turner PE. Phage therapy: a renewed approach to combat antibiotic-resistant bacteria. Cell Host Microbe 2019; 25: 219-232.
15. Lorch A. Bacteriophages: An alternative to antibiotics? Aust Biotechnol 1999; 9: 265-269.
16. Pinto AM, Cerqueira MA, Bañobre-Lópes M, Pastrana LM, Sillankorva S. Bacteriophages for chronic wound treatment: From traditional to novel delivery systems. Viruses 2020; 12: 235.
17. Karn SL, Gangwar M, Kumar R, Bhartiya SK, Nath G. Phage therapy: a revolutionary shift in the management of bacterial infections, pioneering new horizons in clinical practice, and reimagining the arsenal against microbial pathogens. Front Med (Lausanne) 2023; 10: 1209782.
18. Rezaei Z, Elikaei A, Barzi SM, Shafiei M. Isolation, characterization, and antibacterial activity of lytic bacteriophage against methicillin-resistant Staphylococcus aureus causing bedsore and diabetic wounds. Iran J Microbiol 2022; 14: 712-720.
19. Lashtoo Aghaee B, Alikhani MY, van Leeuwen WB, Mojtahedi A, Kazemi S, Karami P. Conventional Treatment of burn wound infections versus phage therapy. Iran J Med Microbiol 2022; 16: 186-196.
20. Torabi LR, Naghavi NS, Doudi M, Monajemi R. Efficacious antibacterial potency of novel bacteriophages against ESBL-producing Klebsiella pneumoniae isolated from burn wound infections. Iran J Microbiol 2021; 13: 678-690.
21. Cerveny KE, DePaola A, Duckworth DH, Gulig PA. Phage therapy of local and systemic disease caused by Vibrio vulnificus in iron-dextran-treated mice. Infect Immun 2002; 70: 6251-6262.
22. Chang H-C, Chen C-R, Lin J-W, Shen G-H, Chang K-M, Tseng Y-H, et al. Isolation and characterization of novel giant Stenotrophomonas maltophilia phage phiSMA5. Appl Environ Microbiol 2005; 71: 1387-1393.
23. Jo SJ, Lee YM, Cho K, Park SY, Kwon H, Giri SS, et al. Standardization of the Agar plate method for bacteriophage production. Antibiotics 2025; 14: 2.
24. Peters DL, Harris G, Davis CM, Dennis JJ, Chen W. Bacteriophage isolation, purification, and characterization techniques against ubiquitous opportunistic pathogens. Curr Protoc 2022; 2(11): e594.
25. Skaradzińska A, Ochocka M, Śliwka P, Kuźmińska-Bajor M, Skaradziński G, Friese A, et al. Bacteriophage amplification–A comparison of selected methods. J Virol Methods 2020; 282: 113856.
26. Lapras B, Marchand C, Merienne C, Medina M, Kolenda C, Laurent F, et al. Rationalisation of the purification process for a phage active pharmaceutical ingredient. Eur J Pharm Biopharm 2024: 203: 114438.
27. Biswas B, Adhya S, Washart P, Paul B, Trostel AN, Powell B, et al. Bacteriophage therapy rescues mice bacteremic from a clinical isolate of vancomycin-resistant Enterococcus faecium. Infect Immun 2002; 70: 204-210.
28. Nikumbh KV, Sevankar SG, Patil MP. Formulation development, in vitro and in vivo evaluation of microemulsion-based gel loaded with ketoprofen. Drug Deliv 2015; 22: 509-515.
29. Jokar J, Abdulabbas HT, Javanmardi K, Mobasher MA, Jafari S, Ghasemian A, et al. Enhancement of bactericidal effects of bacteriophage and gentamicin combination regimen against Staphylococcus aureus and Pseudomonas aeruginosa strains in a mice diabetic wound model. Virus Genes 2024; 60: 80-96.
30. Björn C, Noppa L, Näslund Salomonsson E, Johansson A-L, Nilsson E, Mahlapuu M, et al. Efficacy and safety profile of the novel antimicrobial peptide PXL150 in a mouse model of infected burn wounds. Int J Antimicrob Agents 2015; 45: 519-524.
31. Shrum B, Anantha RV, Xu SX, Donnelly M, Haeryfar SM, McCormick JK, et al. A robust scoring system to evaluate sepsis severity in an animal model. BMC Res Notes 2014; 7: 233.
32. Brans TA, Dutrieux RP, Hoekstra MJ, Kreis RW, du Pont JS. Histopathological evaluation of scalds and contact burns in the pig model. Burns 1994; 20 Suppl 1: S48-S51.
33. Jokar J, Saleh RO, Rahimian N, Ghasemian A, Ghaznavi G, Radfar A, et al. Antibacterial effects of single phage and phage cocktail against multidrug-resistant Klebsiella pneumoniae isolated from diabetic foot ulcer. Virus Genes 2023; 59: 635-642.
34. Jamalludeen N, Shakir D. In vitro effect of newly bacteriophages isolates against Escherichia coli serogroups collected from the local hospital laboratory; their specific characterization and identification. J Microbiol Biotechnol Food Sci 2022; 11(6): e5478.
35. Gupta M, Naik AK, Singh SK. Bacteriological profile and antimicrobial resistance patterns of burn wound infections in a tertiary care hospital. Heliyon 2019; 5(12): e02956.
36. Litwin A, Rojek S, Gozdzik W, Duszynska W. Pseudomonas aeruginosa device associated–healthcare associated infections and its multidrug resistance at intensive care unit of University Hospital: polish, 8.5-year, prospective, single-centre study. BMC Infect Dis 2021; 21: 180.
37. Wyres KL, Lam MMC, Holt KE. Population genomics of Klebsiella pneumoniae. Nat Rev Microbiol 2020; 18: 344-359.
38. Azeredo J, Sutherland IW. The use of phages for the removal of infectious biofilms. Curr Pharm Biotechnol 2008; 9: 261-266.
39. Sharma S, Datta S, Chatterjee S, Dutta M, Samanta J, Vairale MG, et al. Isolation and characterization of a lytic bacteriophage against Pseudomonas aeruginosa. Sci Rep 2021; 11: 19393.
40. Li N, Zeng Y, Bao R, Zhu T, Tan D, Hu B. Isolation and characterization of novel phages targeting pathogenic Klebsiella pneumoniae. Front Cell Infect Microbiol 2021; 11: 792305.
41. Gibran NS, Boyce S, Greenhalgh DG. Cutaneous wound healing. J Burn Care Res 2007; 28: 577-579.
42. Weber-Dabrowska B, Mulczyk M, Górski A. Bacteriophage therapy for infections in cancer patients. Clin Appl Immun Rev 2001; 1: 131-134.
43. Kumari S, Harjai K, Chhibber S. Evidence to support the therapeutic potential of bacteriophage Kpn5 in burn wound infection caused by Klebsiella pneumoniae in BALB/c mice. J Microbiol Biotechnol 2010; 20: 935-941.
44. Finlay‐Jones JJ, Davies KV, Sturm LP, Kenny PA, Hart PH. Inflammatory processes in a murine model of intra‐abdominal abscess formation. J Leukoc Biol 1999; 66: 583-587.
45. Guerra MES, Destro G, Vieira B, Lima AS, Ferraz LFC, Hakansson AP, et al. Klebsiella pneumoniae biofilms and their role in disease pathogenesis. Front Cell Infect Microbiol 2022; 12: 877995.
46. Soothill J. Use of bacteriophages in the treatment of Pseudomonas aeruginosa infections. Expert Rev Anti Infect Ther 2013; 11: 909-915.
47. Kumari S, Harjai K, Chhibber S. Bacteriophage versus antimicrobial agents for the treatment of murine burn wound infection caused by Klebsiella pneumoniae B5055. J Med Microbiol 2011; 60: 205-210.
48. Punjataewakupt A, Napavichayanun S, Aramwit P. The downside of antimicrobial agents for wound healing. Eur J Clin Microbiol Infect Dis 2019; 38: 39-54.
49. Shevchenko OV, Kharina AV, Korniienko NO, Budzanivska IH, Andriichuk OM, Pozhylov IM, et al. Phage therapy in traumatology: A review on perspectives for treating acute wounds and Post-Surgical complications. Mikrobiol Zh 2024; 86: 117-136.
| Files | ||
| Issue | Vol 17 No 1 (2025) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v17i1.17803 | |
| Keywords | ||
| Bacteriophages; Antibiotic resistance; Hydroxy-propyl-methyl cellulose | ||
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How to Cite
1.
Adibi M, Javanmardi K, Ali A.H. Saeed NA-H, Mobasher MA, Jokar J, Ghasemian A, Rahimian N, Soltani Hekmat A. Infected burn wound healing using Hydroxy-propyl-methyl cellulose gel containing bacteriophages against Pseudomonas aeruginosa and Klebsiella pneumoniae. Iran J Microbiol. 2025;17(1):69-79.
