@Article{, title={Use of 820 nm Diode Laser Conjugating With Methylene Blue to Improve Open Wound Healing in Rats}, author={Thaier Alwan Abid ثاير علوان عبد and Roaa Imad Hameed رؤى عماد حميد}, journal={Al-Qadisiyah Journal of Veterinary Medicine Sciences مجلة القادسية لعلوم الطب البيطري}, volume={21}, number={2}, pages={31-47}, year={2022}, 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.

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