Macroscopic and Microscopic Survey of the Comparative Effects of Lactobacillus plantarum 299v, Its Supernatant, and Imipenem on Infectious Burn Wound Healing in Rats

Document Type : Original Research

Authors
S. Soleymanzadeh moghaddam 1
T. Mousavi Shabestari 2
F. Heidari 3
S. Rasouli koohi 2
M. Afshar 2
R. Omranian 2
1 Antimicrobial Resistance Research Center, Institute of Immunology and infection diseases, Iran University of Medical Sciences, Tehran, Iran
2 Antimicrobial Resistance Research Centre, Institute of Immunology and infection diseases, Iran University of Medical Sciences, Tehran, Iran
3 Cellular and Molecular research Center, Department of Anatomy, Qom University of Medical Science, Qom, Iran
10.29252/iem.6.2.85
Abstract
Aims: Burn infections are one of the most common causes of mortality in the world. On the other hand, microbial resistance to antibiotics has caused concern in the medical community. Therefore, controlling burn infections is very important, and using alternative therapies instead of antibiotics could be a good solution to this problem.

Materials & Methods: Lactobacillus plantarum 299v strains were used in the experiments. Fifty male Wistar rats were prepared, and burn was induced in animals. The burn wounds were inoculated with clinical strains of MDR Pseudomonas aeruginosa in all animals and then treated daily with an eucerin ointment containing different compositions, including NaCl, imipenem, probiotic cell pellet, probiotic supernatant, and probiotic cell pellet + probiotic supernatant. The wound healing process was evaluated in animals after 7 days of treatment. Comparisons between different groups were performed using One-way ANOVA and Turkey’s post hoc tests.

Findings: After 7 days of treatment, the mean wound size in the probiotic cell pellet group was significantly lower than in the control and imipenem groups. Also, the mean wound size in the probiotic supernatant group was significantly lower than in the imipenem group. Histological parameters related to skin repair in the probiotic cell pellet group was better than in the control and antibiotic groups. Also, inflammation in the probiotic cell pellet group was less than in the control and imipenem groups.

Conclusion: The macroscopic results of this study supported the microscopic results and showed that the mean size of the burn wounds in the probiotic cell pellet group was less than in the control and imipenem groups after 7 days of treatment.

Keywords

Subjects

1.Rybarczyk MM, Schafer JM, Elm CM, Sarvepalli S, Vaswani PA, Balhara KS, et al. A systematic review of burn injuries in low-and middle-income countries: epidemiology in the WHO-defined African Region. Afr J Emerg Med. 2017;7:30-7.
2. S.M. Tahir, Asadullah Makhdoom, Shazia Awan and Syed Asad Ali .Role of Probiotics in the Management of Burns Patients,World Journal of Medical Sciences . 2014; 11 (3): 417-421.
3. Amir Abbas Barzegari, Masood Hashemzaei, Raheleh Majdani, Ali-Reza Alihemmati. Effects of topical treatment of second-degree burn wounds with Lactobacillus acidophilus on the wound healing process in male rats. Pharm Biomed Res. 2017; 3(3):23.
4. The survey of the comparative effects of Lactobacillus plantarum; in the form of germ, supernatant and the combination of both and imipenem; on infected burn wound healing, Medical Journal of the Islamic Republic of Iran (in press).
5. Church D, Elsayed S, Reid O, Winston B, Lindsay R. Burn wound infections. Clinical microbiology reviews. 2006;19(2):403-34.
6. Japoni A, Farshad S, Alborzi A. Pseudomonas aeruginosa: burn infection, treatment and antibacterial resistance. Iranian Red Crescent Medical Journal. 2009; 3:244-53.
7. Somayeh Soleymanzadeh Moghadam, Zeinab Fagheei Aghmiyuni, Nazanin Mohammad, Ali Anissian, Maryam Azimi, Ali Majidpour,et al. Survey of the Effect of Granulocyte‑colony‑stimulating Factor (G CSF) on Bacterial Translocation and Wound Healing in Burned Mice. Archives of Trauma Research. 2019; 8:149-54.
8. Lee MC, Lin LH, Hung KL, Wu HY. Oral bacterial therapy promotes recovery from acute diarrhea in children. Acta Paediatr Taiwan. 2001;42:301-5.
9. Felgner S, Kocijancic D, Frahm M, Weiss S. Bacteria in cancer therapy: renaissance of an old concept. Int J Microbiol. 2016;2016.
10. Araya M, Morelli L, Reid G, Sanders M, Stanton C, Pineiro M. Joint FAO/WHO Working Group report on drafting guidelines for the evaluation of probiotics in food. London, Canada: World Health Organization. Food and Agriculture Organization of the United Nations 2002.
11. Sekhar MS, Unnikrishnan M, Vijayanarayana K, Rodrigues GS, Mukhopadhyay C. Topical ,
application/formulation of probiotics: Will it be a novel treatment approach for diabetic foot ulcer? Medical Hypotheses. 2014; 82(1): 86-88.
12. Masoud Salaran, Ahmad Oryan, Behrooz Nikahval, Amir Kamali, Mehran Ghaemi, Fatemeh Abbasi-Teshnizi, et al. Topical Application of Lactobacillus Plantarum on Burn Wound Healing in Diabetic Rats. Iranian Journal of Veterinary Surgery. 2019;14(1); Serial No: 30.
13. Yuying Liu , Jane J. Alookaran and J. Marc Rhoads. Probiotics in Autoimmune and Inflammatory Disorders. Nutrients. 2018; 10: 1537
14. Lavasani S, Dzhambazov B, Nouri M, Fåk F, Buske S, Molin G, et al., A novel probiotic mixture exerts a therapeutic effect on experimental autoimmune encephalomyelitis mediated by IL-10 producing regulatory T cells. PLoS One. 2010; 5(2): e9009.

15. Amir Abbas Barzegari, Masood Hashemzaei1, Ali-Reza Alihemmati, Soroush Soltani, Behrouz Naseri. Effects of Lactobacillus rhamnosus (ATCC 7469) ointment on second-degree burn wound in Wistar rat. J Bas Res Med Sci. 2018; 5(1):1-9.
16. Della Riccia D, Bizzini F, Perilli M, Polimeni A, Trinchieri V, Amicosante G, et al. Anti‐inflammatory effects of Lactobacillus brevis (CD2) on periodontal disease. Oral Dis. 2007;13:376-85.
17. Harith Jabbar Fahad Al-Mathkhury and Hind Jabbar Abdul-Rahman Al-Aubeidi. PROBIOTIC EFFECT OF LACTOBACILLI ON MICE INCISIONAL WOUND INFECTIONS. Journal of Al-Nahrain University. 2008; 11(3): 111-116.

18. Guarino A, Canani RB, Spagnuolo MI, Albano F, Di Benedetto L. Oral bacterial therapy reduces the duration of symptoms and of viral excretion in children with mild diarrhea. J Pediatr
Gastroenterol Nutr. 1997; 25:516-9.
19. J. Halper, L. Leshin, S. Lewis, and W. Li. “Wound healing and angiogenic properties of supernatants from Lactobacillus cultures” Exp. Biol. Med. 2003; 228:1329–1337.
20. W. Li, B. Brackett, and J. Halper. “Culture supernatant of Lactobacillus acidophilus stimulates proliferation of embryonic cells”. Exp. Biol. Med. 2005; 230: 494–501.
21. Valdez J, Peral M, Rachid M, Santana M, Perdigon G. Interference of Lactobacillus plantarum with Pseudomonas aeruginosa in vitro and in infected burns: the potential use of probiotics in wound treatment. Clinical Microbiology and Infection. 2005; 11(6): 472-479.

22. Heydari Nasrabadi, Tajabadi Ebrahimi, Dehghan Banadaki, Sh, Torabi Kajousangi, M and Zahedi, F. Study of cutaneous wound healing in rats treated with Lactobacillus plantarum on days 1, 3, 7, 14 and 21. African Journal of Pharmacy and Pharmacology. 2011; 5(21): 2395-2401.
23. Ebrahimi TM, Ouwehand AC, Hejazi MA, Jafari P. Traditional Iranian dairy products: A source of potential probiotic lactobacilli. Afr. J. Microbiol. 2011; 5: 20-27.
24. Goudarzvand M, Rasouli Koohi S, Khodaii Z, Soleymanzadeh Moghadam S. Probiotics Lactobacillus plantarum and bifidobacterium B94: cognitive function in demyelinated model. Med J Islam Repub Iran. 2016; 30:391.

25. Greenhalgh DG, Sprugel KH, Murray MJ, Ross R. PDGF and FGF stimulate wound healing in the genetically diabetic mouse. The American journal of pathology. 1990; 136:1235.
26. Eroglu E, Agalar F, Altuntas I, Eroglu F. Effects of granulocyte-colony stimulating factor on wound healing in a mouse model of burn trauma. The Tohoku journal of experimental medicine. 2004; 204:11-6.
27. Yoram Abramov,Barbara Golden, Megan Sullivan, Sylvia M. Botros, Jay-James R. Miller, Adeeb Alshahrour, et.al. Histologic characterization of vaginal vs. abdominal surgical wound healing in a rabbit model. Wound Rep Reg. 2007; 15: 80–86.
28. David G. Greenhalgh, Katherine H. Spruge, Mark J. Murray, and Russe Rosst. PDGF and FGF Stimulate Wound Healing in the Genetically Diabetic Mouse. American Journal of Pathology. 1990; 136(6)
29. Hansen JE, Jespersen LK. Wound or tissue dressing comprising lactic acid bacteria. US Patent 0143447 A1, 10 June 2010
30. Prince T, McBain AJ, O'Neill CA. Lactobacillus reuteri protects epidermal keratinocytes from Staphylococcus aureus-induced cell death by competitive exclusion. Appl Environ Microbiol. 2012;78:5119-26.
31. Peral MC, Huaman Martinez MA, Valdez JC. Bacteriotherapy with Lactobacillus plantarum in burns. Int Wound J. 2009; 6:73-81.
32. Carol Nhan, Aren Bezdjian, Shyamali Saha, Satya Prakash, Lily H. P. Nguyen and Sam J. Danie. Safety of transtympanic application of probiotics in a chinchilla animal model. Journal of Otolaryngology - Head and Neck Surgery. 2017; 46:63.
33. Simova E, Beshkova D, Angelov A, Hristozova T, Frengova G, Spasov Z. Lactic acid bacteria and yeasts in kefir grains and kefir made from them. Journal of Industrial Microbiology and Biotechnology. 2002; 28(1): 1-6.
34. Isolauri E, Kirjavainen P, Salminen S. Probiotics: a role in the treatment of intestinal infection and inflammation? Gut 2002;50:54-9.
35. Raffaella Campana, Saskia van Hemert, and Wally Baffone. Strain‑specific probiotic properties of lactic acid bacteria and their interference with human intestinal pathogens invasion.
Gut Pathog. 2017; 9:12
36. Sharma J, Chauhan D. Inhibition of Pseudomonas aeruginosa by antibiotics
and probiotics combinations-In vitro study. Europ J Exp Biol. 2014;4(6):10–4.
37. Guilfoile P, Alcamo IE. Antibiotic-resistant bacteria: Infobase Publishing. 2007.
38. Ammann AJ, Beck LS, DeGUZMAN L, Hirabayashi SE, Pun Lee W, McFATRIDGE L, et al. Transforming Growth Factor‐β Effect on Soft Tissue Repair. Annals of the New York Academy of Sciences. 1990; 593(1): 124-134.
39. Kekkonen RA, Lummela N, Karjalainen H, Latvala S, Tynkkynen S, Järvenpää S, et al. Probiotic intervention has strain-specific anti-inflammatory effects in healthy adults. World JGastroenterol. 2008;14:2029-36.

40. von Schillde MA, Hörmannsperger G, Weiher M, Alpert CA, Hahne H, Bäuerl C, et al. Lactocepin secreted by Lactobacillus exerts anti-inflammatory effects by selectively degrading
proinflammatory chemokines. Cell Host Microbe. 2012;11:387- 96.
Infection Epidemiology and Microbiology
Volume 6, Issue 2
June 2020
Pages 85-93