Volume 29, Issue 4 (December 2025)
Physiol Pharmacol 2025, 29(4): 408-423 |
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Mousavinezhad S A, Harzandi N, Marjani A, Jafari P. Evaluation of the effect of biodegradable dressing prepared from chitosan and external microvesicles of Bifidobacterium bifidum on burn wound healing in male Wistar rats. Physiol Pharmacol 2025; 29 (4) :408-423
URL: http://ppj.phypha.ir/article-1-2417-en.html
URL: http://ppj.phypha.ir/article-1-2417-en.html
Abstract: (1907 Views)
Introduction: The burn wound healing process is multi-variable, and various factors are involved. Chitosan-based dressings are acceptable materials for wound healing. PProbiotics and postbiotics can in-crease the effectiveness of these dressings. This study aimed to investigate the effect of a biodegradable dressing prepared from chitosan and external microvesicles of Bifidobacterium bifidum on burn wound healing in male Wistar rats.
Methods: Bifidobacterium bifidum was cultured, the activated crude supernatant (ACS) was separated, and the microvesicles were isolated with the help of Ultra-centrifugation at 150,000×g at 4°C for 30 minutes. Chitosan films containing and without microvesicles were prepared using a heater stirrer at 29°C. The effectiveness of the membranes was evaluated. For this purpose, 60 male rats with second-degree burns were randomly divided into 5 groups (n=12) and were treated for 21 days. The process of wound healing was examined macroscopically and microscopically (wound histology and evaluation of gene expression of cytokines interleukin-8, interleukin-10, and VEGF) on the days 3, 7, 14 and 21. The obtained data were analyzed by IBM SPSS.21 software and using Kolmogorov-Smirnov, Kruskal-Wallis, and Mann-Whitney tests with p ≤ 0.05.
Results: The average diameter of microvesicles was 174 ± 52nm. The thickness of the layers was 2mm. The wounds of the group post-biotic and positive control were closed on the 14th day. Collagen production and epithelialization, as well as inflammation control in the treatment groups, were higher than negative control.
Conclusion: The use of chitosan membranes, particularly those enhanced with ACS and microvesicles, represents a significant advancement in wound care by inflammation control, offering a promising strategy for improving the healing of burn wounds.
Methods: Bifidobacterium bifidum was cultured, the activated crude supernatant (ACS) was separated, and the microvesicles were isolated with the help of Ultra-centrifugation at 150,000×g at 4°C for 30 minutes. Chitosan films containing and without microvesicles were prepared using a heater stirrer at 29°C. The effectiveness of the membranes was evaluated. For this purpose, 60 male rats with second-degree burns were randomly divided into 5 groups (n=12) and were treated for 21 days. The process of wound healing was examined macroscopically and microscopically (wound histology and evaluation of gene expression of cytokines interleukin-8, interleukin-10, and VEGF) on the days 3, 7, 14 and 21. The obtained data were analyzed by IBM SPSS.21 software and using Kolmogorov-Smirnov, Kruskal-Wallis, and Mann-Whitney tests with p ≤ 0.05.
Results: The average diameter of microvesicles was 174 ± 52nm. The thickness of the layers was 2mm. The wounds of the group post-biotic and positive control were closed on the 14th day. Collagen production and epithelialization, as well as inflammation control in the treatment groups, were higher than negative control.
Conclusion: The use of chitosan membranes, particularly those enhanced with ACS and microvesicles, represents a significant advancement in wound care by inflammation control, offering a promising strategy for improving the healing of burn wounds.
Type of Manuscript: Experimental research article |
Subject:
Clinical Physiology/Pharmacology
References
1. Ahn C-B, Jung W-K, Park S-J, Kim Y-T, Kim W-S, Je J-Y. Gallic acid-g-chitosan modulates inflammatory responses in LPS-stimulated RAW264. 7 cells via NF-κB, AP-1, and MAPK pathways. Inflammation 2016; 39: 366-374. [DOI:10.1007/s10753-015-0258-2]
2. Al-Nedawi K, Mian M F, Hossain N, Karimi K, Mao Y K, Forsythe P, et al. Gut commensal microvesicles reproduce parent bacterial signals to host immune and enteric nervous systems. The FASEB Journal 2015; 29: 684-695. [DOI:10.1096/fj.14-259721]
3. Babbitt D M, Diaz K M, Feairheller D L, Sturgeon K M, Perkins A M, Veerabhadrappa P, et al. Endothelial activation microparticles and inflammation status improve with exercise training in African Americans. International Journal of Hypertension 2013; 2013: 538017. [DOI:10.1155/2013/538017]
4. Barrientos S, Stojadinovic O, Golinko M S, Brem H, Tomic-Canic M. Growth factors and cytokines in wound healing. Wound Repair and Regeneration 2008; 16: 585-601. [DOI:10.1111/j.1524-475X.2008.00410.x]
5. Bazjou A, Jafari P, Marjani A, Akbari N. Effect of cell-free supernatant of bifidobacterium bifidum combined with chitosan biodegradable film on full thickness wound healing in rats. Physiology and Pharmacology 2022; 26: 468-479.
6. Brown L, Kessler A, Cabezas-Sanchez P, Luque-Garcia J L, Casadevall A. Extracellular vesicles produced by the G ram-positive bacterium B acillus subtilis are disrupted by the lipopeptide surfactin. Molecular Microbiology 2014; 93: 183-198. [DOI:10.1111/mmi.12650]
7. Cai E Z, Ang C H, Raju A, Tan K B, Hing E C H, Loo Y, et al. Creation of consistent burn wounds: a rat model. Archives of Plastic Surgery 2014; 41: 317-324. [DOI:10.5999/aps.2014.41.4.317]
8. Ceylan H G, Atasoy A F. Lactobacillus rhamnosus gg ve bifidobacterium bifidum bb-12 içeren yenilebilir filmlerin karakterizasyonu. Adıyaman Üniversitesi Mühendislik Bilimleri Dergisi 2022; 9: 194-203. [DOI:10.54365/adyumbd.1073615]
9. Choi Y J, Shin H S. Antibacterial effect of eight probiotic strains of bifidobacterium against pathogenic Staphylococcus aureus and Pseudomonas aeruginosa. Journal of Bacteriology and Virology 2021; 51: 128-137. [DOI:10.4167/jbv.2021.51.3.128]
10. Diomede F, Marconi G D, Fonticoli L, Pizzicanella J, Merciaro I, Bramanti P, et al. Functional relationship between osteogenesis and angiogenesis in tissue regeneration. International Journal of Molecular Sciences 2020; 21: 3242. [DOI:10.3390/ijms21093242]
11. DiPietro L A. Angiogenesis and wound repair: when enough is enough. Journal of Leucocyte Biology 2016; 100: 979-984. [DOI:10.1189/jlb.4MR0316-102R]
12. Domínguez Rubio A P, Martínez J H, Martínez Casillas D C, Coluccio Leskow F, Piuri M, Pérez O E. Lactobacillus casei BL23 produces microvesicles carrying proteins that have been associated with its probiotic effect. Frontiers in Microbiology 2017; 8: 1783. [DOI:10.3389/fmicb.2017.01783]
13. Dong Y, Cui M, Qu J, Wang X, Kwon S H, Barrera J, et al. Conformable hyaluronic acid hydrogel delivers adipose-derived stem cells and promotes regeneration of burn injury. Acta Biomaterialia 2020; 108: 56-66. [DOI:10.1016/j.actbio.2020.03.040]
14. Falanga V. Wound healing and its impairment in the diabetic foot. The Lancet 2005; 366: 1736-1743. [DOI:10.1016/S0140-6736(05)67700-8]
15. Ghaneialvar H, Arjmand S, Sahebghadam Lotfi A, Soleimani M, Mashhadi Abbas F. Influence of adipose derived mesenchymal stem cells on the effective inflammatory factors of diabetic wound healing in animal models. Journal of Mazandaran University of Medical Sciences 2017; 27: 12-21.
16. Grazul-Bilska A T, Johnson M L, Bilski J J, Redmer D A, Reynolds L P, Abdullah A, et al. Wound healing: the role of growth factors. Drugs Today (Barc) 2003; 39: 787-800. [DOI:10.1358/dot.2003.39.10.799472]
17. Guo S a, DiPietro L A. Factors affecting wound healing. Journal of Dental Research 2010; 89: 219-229. [DOI:10.1177/0022034509359125]
18. Ilyas S. Immunohistochemistry examination to reveal the pathogenesis of Oral Squamous Cell Carcinoma. International Journal of Ecophysiology 2022; 4: 5-25. [DOI:10.32734/ijoep.v4i1.11150]
19. Jamaran S, Jafari P, Marjani A, Akbari N, Feizabad M M. Novel wound dressing based on postbiotic/chitosan film accelerates cutaneous wound healing. Jundishapur Journal of Microbiology 2021; 14. [DOI:10.5812/jjm.120806]
20. Jara C P, Carraro R S, Zanesco A, Andrade B, Moreira K, Nogueira G, et al. A smartphone app for individual xylazine/ketamine calculation decreased anesthesia-related mortality in mice. Frontiers in Veterinary Science 2021; 8: 651202. [DOI:10.3389/fvets.2021.651202]
21. Kambouris A R, Brammer J A, Roussey H, Chen C, Cross A S. A combination of burn wound injury and Pseudomonas infection elicits unique gene expression that enhances bacterial pathogenicity. mBio2023; 14: e0245423. [DOI:10.1128/mbio.02454-23]
22. Kojima K, Okamoto Y, Kojima K, Miyatake K, Fujise H, Shigemasa Y, et al. Effects of chitin and chitosan on collagen synthesis in wound healing. Journal of Veterinary Medical Science 2004; 66: 1595-1598. [DOI:10.1292/jvms.66.1595]
23. Kouitcheu Mabeku L B, Ngue S, Bonsou Nguemo I, Leundji H. Potential of selected lactic acid bacteria from Theobroma cacao fermented fruit juice and cell-free supernatants from cultures as inhibitors of Helicobacter pylori and as good probiotic. BMC Research Notes 2020; 13: 1-7. [DOI:10.1186/s13104-020-4923-7]
24. Li D, Zhang C, Song F, Lubenec I, Tian Y, Song Q. VEGF regulates FGF-2 and TGF-β1 expression in injury endothelial cells and mediates smooth muscle cells proliferation and migration. Microvascular Research 2009; 77: 134-142. [DOI:10.1016/j.mvr.2008.09.007]
25. McFadyen J D, Kaplan Z S. Platelets are not just for clots. Transfusion Medicine Reviews 2015; 29: 110-119. [DOI:10.1016/j.tmrv.2014.11.006]
26. Mercandetti M, Cohen A. Wound healing and repair. Emedicine 2017; 14: 12-20.
27. Mirzaian F, Amirizadeh N, Nikougoftar Zarif M, Atarodi K, Nakhlestani M. Isolation and characterization of Placenta-MSC derived microvesicles. Scientific Journal of Iran Blood Transfus Organ 2015; 12: 135-142.
28. Miyagi M, Uchida K, Takano S, Fujimaki H, Aikawa J, Sekiguchi H, et al. Macrophage-derived inflammatory cytokines regulate growth factors and pain-related molecules in mice with intervertebral disc injury. Journal of Orthopaedic Research® 2018; 36: 2274-2279. [DOI:10.1002/jor.23888]
29. Mohite P, Rahayu P, Munde S, Ade N, Chidrawar V R, Singh S, et al. Chitosan-based hydrogel in the management of dermal infections: a review. Gels 2023; 9: 594. [DOI:10.3390/gels9070594]
30. Mousavi Nezhad S A, Tajabadi Ebrahimi M, Zilabi R. The Effect of Iran probiotic fermented milk beverage on cholesterol and liver function biomarkers of rat. Journal of Police Medicine 2015; 4: 49-56.
31. Mousavinezhad S A, Harzandi N, Marjani A, Jafari P. Effect of culture supernatant of three probiotic bacterial strains of Bifidobacterium on Pseudomonas aeruginosa. EBNESINA 2023; 25: 25-32.
32. Mousavinezhad S A, Harzandi N, Marjani A, Jafari P. The effectiveness of chitosan wound dressing in the form of a film compared to its hydrogel on burn wound healing in the male wistar rat model. Journal of Police Medicine 2022; 11: 1-17.
33. Naz I, Fatima A, Alhewairini S S, Rehman A. Assessment of antibacterial activity of Syzygium aromaticum extracts, antibiotics and silver sulphadiazine ointment against pathogenic bacteria isolated from the burned and unburned skin. Malaysian Journal of Microbiology 2021; 17. [DOI:10.21161/mjm.200895]
34. Oertelt-Prigione S. The influence of sex and gender on the immune response. Autoimmunity Reviews 2012; 11: A479-A485. [DOI:10.1016/j.autrev.2011.11.022]
35. Rodrigues KL, Caputo LRG, Carvalho JCT, Evangelista J, Schneedorf J M. Antimicrobial and healing activity of kefir and kefiran extract. International Journal of Antimicrobial Agents 2005; 25: 404-408. [DOI:10.1016/j.ijantimicag.2004.09.020]
36. Sabahi S, Homayouni Rad A, Aghebati-Maleki L, Sangtarash N, Ozma M A, Karimi A, et al. Postbiotics as the new frontier in food and pharmaceutical research. Critical Reviews in Food Science and Nutrition 2023; 63: 8375-8402. [DOI:10.1080/10408398.2022.2056727]
37. Sanders ME. How do we know when something called “probiotic” is really a probiotic? A guideline for consumers and health care professionals. Functional Food Reviews 2009; 1: 3-12.
38. Savadogo A, Ouattara AC, Bassole HI, Traore SA. Bacteriocins and lactic acid bacteria-a minireview. African Journal of Biotechnology 2006; 5.
39. Sinha M, Sen CK, Singh K, Das A, Ghatak S, Rhea B, et al. Direct conversion of injury-site myeloid cells to fibroblast-like cells of granulation tissue. Nature Communications 2018; 9: 936. [DOI:10.1038/s41467-018-03208-w]
40. Sorg H, Tilkorn D J, Hager S, Hauser J, Mirastschijski U. Skin wound healing: an update on the current knowledge and concepts. European Surgical Research 2017; 58: 81-94. [DOI:10.1159/000454919]
41. Touré R, Kheadr E, Lacroix C, Moroni O, Fliss I. Production of antibacterial substances by bifidobacterial isolates from infant stool active against Listeria monocytogenes. Journal of Applied Microbiology 2003; 95: 1058-1069. [DOI:10.1046/j.1365-2672.2003.02085.x]
42. Volk SW, Bohling MW. Comparative wound healing-are the small animal veterinarian’s clinical patients an improved translational model for human wound healing research? Wound Repair and Regeneration 2013; 21: 372-381. [DOI:10.1111/wrr.12049]
43. Wang D, Zhang N, Meng G, He J, Wu F. The effect of form of carboxymethyl-chitosan dressings on biological properties in wound healing. Colloids and Surfaces B: Biointerfaces 2020; 194: 111191. [DOI:10.1016/j.colsurfb.2020.111191]
44. Wang F, Cerione RA, Antonyak MA. Isolation and characterization of extracellular vesicles produced by cell lines. STAR Protocols 2021; 2: 100295. [DOI:10.1016/j.xpro.2021.100295]
45. Xu X, Wang R, Sun Z, Wu R, Yan W, Jiang Q, et al. Trehalose enhances bone fracture healing in a rat sleep deprivation model. Annals of Translational Medicine 2019; 7. [DOI:10.21037/atm.2019.05.73]
46. Yin M, Wan S, Ren X, Chu C-C. Development of inherently antibacterial, biodegradable, and biologically active chitosan/pseudo-protein hybrid hydrogels as biofunctional wound dressings. ACS Applied Materials & Interfaces 2021; 13: 14688-14699. [DOI:10.1021/acsami.0c21680]
47. Yoshikawa Y, Morita M, Hosomi H, Tsuneyama K, Fukami T, Nakajima M, et al. Knockdown of superoxide dismutase 2 enhances acetaminophen-induced hepatotoxicity in rat. Toxicology 2009; 264: 89-95. [DOI:10.1016/j.tox.2009.07.017]
48. Zhao J, Li J, Xu S, Feng P. Emerging roles of protein deamidation in innate immune signaling. J Virol 2016; 90: 4262-4268. [DOI:10.1128/JVI.01980-15]
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