Al-Qadisiyah Journal of Veterinary Medicine Sciences

Use of 820 nm Diode Laser Conjugating With Methylene Blue to Improve Open Wound Healing in Rats

Document Type : Research Paper

Authors

Al-Qadisiyah Journal of Veterinary Medicine Sciences

10.29079/qjvms.2022.179317

Abstract

Objective: The study was aimed to evaluate the use of 820 nm diode laser conjugating with methylene blue (MB) to improve open wound healing in rats. Methods: Sixty adult male Wistar rats weighing (240±20gm), aged between (7-12) weeks were divided into four equal groups (n 15); G1/Control group which not receive any treatment. G2/Laser treatment group, were treated with 8J/cm²/day for 7 successive days. G3/ MB treatment group, were treated with topical application of MB dye 0.01% for 5 min/day for 7 successive days. G4/ MB + Laser treatment group were treated with MB and laser for 7 successive days, the session include a topical application of the MB as in G3 followed by laser treatment as in G2. After general anesthesia, two open circular full-thickness (0.8 cm in diameter) skin incision on the back of the animals were made. In treated groups one incision was treated and the 2nd were left as control in the same animal. Assessment of the wound healing was done grossly and microscopically. Results: Macroscopically were seen significant P < 0.05 decreased of the size of wound in treated groups in compare with control at 7, and 14 days, while the readings become near each other at 21 days. The epithelization period in treated groups were 8-10 days and 8 days in control group. Microscopically G1 show no epithelialization, new collagen fiber, new blood vessels, numerous PMNCs, and large number of fibroblasts at 7th day PW. At 14th day exhibit thin epithelial layers connecting the two edges of wound, and more collagen fibers arranged horizontally in the dermis, while at 21st day, no inflammatory cells, more collagen fiber, and less cellular field appear. G2 at 7th day PW, show severe numbers of PMNCs, new collagen fibers, plentiful numbers of fibroblast, and severe numbers of new blood vessels. At 14th day PW, thick well differentiated epithelial layers were seen, upon new collagen fibers, less numbers of PMNCs, plentiful numbers of fibroblast and myofibroblasts. At 21st day, thick even epithelial layers were seen upon mature well-arranged collagen fibers, scanty blood vessels, and myofibroblasts. Buds of hair follicles were seen also. G3 at 7th day PW, show thin epithelial layers, plenty of PMNCs, and numerous new blood vessels with more new collagen fibers. At 14th day PW moderate thickening of epidermis were seen upon deep area of granulation tissue) more collagen and less cellular). At 21st day PW, thin epidermal layers were seen upon mature granulation tissue. G4 at 7th day PW show complete well re-epithelialization. Epidermal layers were seen riding on distinct basement membrane. The dermis seen have dense cellular pinkly granulation tissue, abundant PMNCs and fibroblast and a lot of number of new blood vessels. In deep dermis, scanty sebaceous glands, solitary hair follicles in different stages of development were seen. At 14th day PW normal conventional appearance of the epidermis were seen. The dermis consist of mature granulation tissue with a lot of numbers of fibroblast, myofibroblasts, and little PMNCs. At 21st day PW normal thickness of epidermis were seen resting on basement membrane has many inner invaginations toward the dermis. The dermis was full with mature granulation tissue with no inflammatory cells, no new blood vessels, the collagen fibers were dense thick and regular, more hair follicles and more sebaceous glands were seen.Conclusions: In conclusion, laser treatment with 820 nm were seen improve the healing process and accelerate the proliferation, wound contraction, maturation and remodeling phases of wound healing. Conjugating methylene blue with laser give better enhancement.

 

Keywords

References
  1. Kolarsick PAJ, Kolarsick MA, Goodwin C. Anatomy and Physiology of the Skin. In: Muehlbauer P, McGowan C, editors. Site-Specific Cancer Series: Skin Cancer. Oncology Nursing Society; 2011. p. 1-11.
  2. Broughton G, Janis JE, Attinger CE. Wound Healing: An Overview. Plast Reconstr Surg. 2006;117(1e-S):1e-S-32eS.https://doi.org/10.1097/01.prs.0000222562.60260.f9
  3. Monaco JL, Lawrence WT. Acute wound healing: an overview. Clin Plast Surg. 2003;30:1-12.https://doi.org/10.1016/S0094-1298(02)00070-6
  4. Velnar T, Bailey T, Smrkolj V. The Wound Healing Process: An Overview of the Cellular and Molecular Mechanisms. J Int Med Res. 2009;37(5):1528-42.https://doi.org/10.1177/147323000903700531
  5. Li J, Chen J, Kirsner R. Pathophysiology of acute wound healing. Clin Dermatol. 2007;25:9-18.https://doi.org/10.1016/j.clindermatol.2006.09.007
  6. Atiyeh BS, Ioannovich J, Al-Amm CA, El-Musa KA. Management of Acute and Chronic Open Wounds: The Importance of Moist Environment in Optimal Wound Healing. Curr Pharm Biotechnol. 2002;3:179-95.https://doi.org/10.2174/1389201023378283
  7. Norman G, Dumville JC, Mohapatra DP, Owens GL, Crosbie EJ. Antibiotics and antiseptics for surgical wounds healing by secondary intention. Cochrane Database Syst Rev. 2016;(3):CD011712.https://doi.org/10.1002/14651858.CD011712
  8. Avci P, Gupta A, Sadasivam M, Vecchio D, Pam Z, Pam N, et al. Low-level laser (light) therapy (LLLT) in skin: stimulating, healing, restoring. Semin Cutan Med Surg. 2013 Mar;32(1):41-52.
  9. Diwan MF, Abid TA. The effect of different doses of 660 nm laser therapy on open skin wound healing in Wistar rats. Kufa J Vet Med Sci. 2016;7:130-7.https://doi.org/10.36326/kjvs/2016/v7i24340
  10. Peplow PV, Chung TR, Baxter GD. Photodynamic modulation of wound healing. A review of human and animal studies. Photomed Laser Surg. 2012;30(3):118-48.https://doi.org/10.1089/pho.2011.3142
  11. Hamblin MR, Sabino CP. Systemic Effects. In: Sellera FP, Nascimento CL, Ribeiro MS, editors. Photodynamic Therapy in Veterinary Medicine: From Basics to Clinical Practice. Springer International Publishing Switzerland; 2016. p. 73.https://doi.org/10.1007/978-3-319-45007-0_6
  12. de Mello Pinto VM, et al. Study of Effects of Photodynamic Therapy (PDT), in Scar-Induced Skin Wounds in Rats Wistar: The New Clinical Perspective for Ulcers. Mod Res Inflamm. 2017;6:1-8.https://doi.org/10.4236/mri.2017.61001
  13. Nesi-Reis V, Lera-Nonose DSS, Oyama J, Silva-Lalucci MPP, Demarchi IG, Aristides SMA, et al. Contribution of photodynamic therapy in wound healing: A systematic review. Photodiagn Photodyn Ther. 2018;21:294-305.https://doi.org/10.1016/j.pdpdt.2017.12.015
  14. Kwiatkowski S, Knap B, Przystupski D, Saczko J, Kędzierska E, Knap-Czop K, et al. Photodynamic therapy mechanisms, photosensitizers and combinations. Biomed Pharmacother. 2018;106:1098-107.https://doi.org/10.1016/j.biopha.2018.07.049
  15. Allison RR, Downie GH, Cuenca R, Hu XH, Childs CJH, Sibata CH. Photosensitizers in clinical PDT. Photodiagn Photodyn Ther. 2004;1:27-42.https://doi.org/10.1016/S1572-1000(04)00007-9
  16. Ormand AB, Freeman HS. Dye sensitizers for photodynamic therapy. Materials. 2013;6:817-40.https://doi.org/10.3390/ma6030817
  17. Trevisana E, Menegazzia R, Zabucchia G, Troianb B, Stefano Pratob S, Vitaa F, et al. Effect of methylene blue photodynamic therapy on human neutrophil functional responses. J Photochem Photobiol B Biol. 2019;199:111605.https://doi.org/10.1016/j.jphotobiol.2019.111605
  18. Tardivoa J, Giglio A, Oliveirab C, Gabrielli D, Junqueirab H, Tadab D, et al. Methylene blue in photodynamic therapy: From basic mechanisms to clinical applications. Photodiagn Photodyn Ther. 2005;2:175-91.https://doi.org/10.1016/S1572-1000(05)00097-9
  19. Tedesco A, Jesus P. Low level energy photodynamic therapy for skin processes and regeneration. In: Tanaka Y, editor. Photomedicine - Advances in Clinical Practice. IntechOpen; 2017. p. 75-94.https://doi.org/10.5772/65344
  20. Garcia JGM. The role of photodynamic therapy in wound healing and scarring in human skin. [PhD Thesis]. University of Manchester; 2015.
  21. Garcia VG, de Lima MA, Okamoto T, Milanezi LA, Júnior ECG, Fernandes LA, et al. Effect of photodynamic therapy on the healing of cutaneous third-degree-burn: histological study in rats. Lasers Med Sci. 2010;25(2):221-8.https://doi.org/10.1007/s10103-009-0694-z
  22. Sperandio FF, Simoes A, Aranha ACC, Correa L, Machado de Sousa SCO. Photodynamic Therapy Mediated by Methylene Blue Dye in Wound Healing. Photomed Laser Surg. 2010;28(2):581-7.https://doi.org/10.1089/pho.2009.2601
  23. Oyama J, Fernandes HRMÁ, Lopes Lera-Nonose D, Nesi-Reis V, Galhardo DI, Alessi Aristides S, et al. Photodynamic therapy in wound healing in vivo: a systematic review. Photodiagn Photodyn Ther. 2020;30:101682.https://doi.org/10.1016/j.pdpdt.2020.101682
  24. Gaertner DJ, Hallman TM, Hankenson FC, Batchelder MA. Anesthesia and Analgesia for Laboratory Rodents. In: Fish RE, Brown MJ, Danneman PJ, Karas AZ, editors. Anesthesia and Analgesia in Laboratory Animals. 2nd ed. Elsevier Inc.; 2008. p. 239-98.https://doi.org/10.1016/B978-012373898-1.50014-0
  25. Pastar I, Liang L, Sawaya AP, Wikramanayake TC, Glinos GD, Drakulich S, et al. Preclinical models for wound-healing studies. In: Marques AP, Pirraco RP, Reis RL, editors. Skin Tissue Models. 1st ed. Elsevier; 2018. p. 223-53.https://doi.org/10.1016/B978-0-12-810545-0.00010-3
  26. Estevao LD, Vieira P, Leite AGB, Bulhoes AV, Barcelos LS, Neto J. Morphological Evaluation of Wound Healing Events in the Excisional Wound Healing Model in Rats. Bio-protocol. 2019;9(13):e3285.https://doi.org/10.21769/BioProtoc.3285
  27. Sardari K, Dehgan MM, Mohri M, Emami MR, Mirshahi A, Maleki M, et al. Macroscopic aspects of wound healing (contraction and epithelialization) after topical administration of allicin in dogs. Comp Clin Pathol. 2006;15:231-5.https://doi.org/10.1007/s00580-006-0634-2
  28. Patil MVK, Kandhare AD, Bhise SD. Pharmacological evaluation of ethanolic extract of Daucus carota Linn root formulated cream on wound healing using excision and incision wound model. Asian Pac J Trop Biomed. 2012;S646-55.https://doi.org/10.1016/S2221-1691(12)60290-1
  29. Nagar HK, Srivastava AK, Srivastava R, Kurmi ML, Chandel HS, Ranawat MS. Pharmacological Investigation of the Wound Healing Activity of Cestrum nocturnum (L.) Ointment in Wistar Albino Rats. J Pharmaceutics. 2016;2016:9249040.https://doi.org/10.1155/2016/9249040
  30. Belachew TF, Asrade S, Geta M, Fentahun E. In Vivo Evaluation of Wound Healing and Anti-Inflammatory Activity of 80% Methanol Crude Flower Extract of Hagenia abyssinica (Bruce) J.F. Gmel in Mice. Evid Based Complement Alternat Med. 2020;2020:9645792.https://doi.org/10.1155/2020/9645792
  31. Luna IG. Manual of histology Staining methods of the armed force institute & pathology. 3rd ed. New York: McGraw-Hill; 1968. p. 34-215.
  32. Tabakoglu HO, Sani MM, Uba AI, Abdullahi UA. Assessment of circular wound healing in rats after exposure to 808-nm laser pulses during specific healing phases. Lasers Surg Med. 2016;48(4):409-15.https://doi.org/10.1002/lsm.22462
  33. Petersen H, Tavakoli F, Kruber S, Münscher A, Gliese A, Hansen NO, et al. Comparative study of wound healing in rat skin following incision with a novel picosecond infrared laser (PIRL) and different surgical modalities. Lasers Surg Med. 2016;48(4):385-91.https://doi.org/10.1002/lsm.22498
  34. Silva JCE, Lacava ZGM, Kuckelhaus S, Silva LP, Neto LFM, Sauro EE, et al. Evaluation of the use of low level laser and photosensitizer drugs in healing. Lasers Surg Med. 2004;34(5):451-7.https://doi.org/10.1002/lsm.20062
  35. Woo KY, Heil J. A prospective evaluation of methylene blue and gentian violet dressing for management of chronic wounds with local infection. Int Wound J. 2017;14(6):1029-35.https://doi.org/10.1111/iwj.12753
  36. Moreira SH, Pazzini JM, Álvarez JLG, et al. Evaluation of angiogenesis, inflammation, and healing on irradiated skin graft with low-level laser therapy in rats (Rattus norvegicus albinus wistar). Lasers Med Sci. 2020;35:1103-9.https://doi.org/10.1007/s10103-019-02917-y
  37. Giuliani A, Lorenzini L, Gallamini M, Massella A, Giardino L, Calzà L. Low infrared laser light irradiation on cultured neural cells: effects on mitochondria and cell viability after oxidative stress. BMC Complement Altern Med. 2009;9(1).https://doi.org/10.1186/1472-6882-9-8
  38. Huang YY, Chen ACH, Carroll JD, Hamblin MR. Biphasic Dose Response in Low Level Light Therapy. Dose-Response. 2009;7(4).https://doi.org/10.2203/dose-response.09-027.Hamblin
Al-Qadisiyah Journal of Veterinary Medicine Sciences
Volume 21, Issue 2
June 2022
Pages 31-47
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