The Effect of Topical Nitroglycerin on Wound Healing in Rabbits using Histopathological Investigation
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Abstract
Introduction: Nitroglycerin can increase the Cycle Guanosine Mono Phosphate level, enhance nitric oxide rate in tissues dilate vessels, and intensify perfusion within tissues. The aim of the present study was to conduct a microscopic investigation addressing the effect of topical 2% Nitroglycerin ointment on wound healing in rabbits.
Materials and methods: Six adult male New Zealand white rabbits, weighing approximately 2.25 kg were used. Two wounds were created on each side of the spinal column. The wounds reached the deep fascia and their dimensions were 15 15 mm. The left wound was used as the control and the right wound was used as the experimental one. Immediately after the creation of wounds, a layer of nitroglycerin 2% with 1 mm thickness was put on the experimental wound daily for seven days. On days 3, 5, 7, and 14 after cutting, both the experimental and control wounds with a margin of healthy tissues were taken for the histopathological examination.
Results: The distance of the two edges on the wound in treated wounds reached a significant difference, compared with control wounds on day 14. The number of inflammatory cells (with neutrophils format) in the treatment group was significantly less than those of the control group starting from day 5. In addition, the fibrin clot diameter in the treatment group was significantly less than the control group on days 5, 7, and 14. At the beginning of the experiment angiogenesis in the control group was more than in the treatment group, but it was the same in both groups on day 7, and angiogenesis in the treatment group was more than in the control group after day 7. The volume of granulation in the treatment group was more than control group and there was a significant difference on days 5, 7, and 14. Epithelial tissues diameter was higher in the treatment group and the difference became significant on day 14.
Conclusion: In conclusion, the findings indicated a promising function of topical NTG in wound healing of anal fissures, tendinopathies, CNH, diabetic foot, or skin flap necrosis.
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References
Brueckner CS, Becker MO, Kroencke T, Huscher D, Scherer HU, Worm M, Burmester G, and Riemekasten G. Effect of sildenafil on digital Ulcers in systemic sclerosis: analysis from a single centre pilot
study. Ann Rheum Dis. 2010; 60: 1475-1478. DOI: https://www.doi.org/10.1136/ard.2009.116475
Cross SE. An experimental model to investigate the dynamics of wound contraction. Br J Plastic surgery. 1995; 58: 189-197. DOI: https://www.doi.org/10.1016/0007-1226(95)90001-2
Salcido R. Wound Healing the NO Connection, advances in Skin and wound care. J Prevention and Healing. 2008; 21: 106-109. DOI: https://www.doi.org/10.1097/01.ASW.0000305426.55671.42
Majure DT, Hallaux M, Yeghiazarians Y, and Boyle AJ. Topical nitroglycerin and lidocaine locally vasodilate the radial artery without affecting systemic blood pressure: a dose-finding phase I study. J Crit Care. 2012; 27: P532. DOI: https://www.doi.org/10.1016/j.jcrc.2012.04.019
Pipelzadeh M H, and et al. A study on the effect of modulation of intracellular calcium on excisional wound healing in rabbit. Iranian Biomedical J. 2003; 7: 161-166. Available at: http:// ibj.pasteur.ac.ir/article-1-523-en.pdf
Ignarro LJ, Cirino G, Casini A, and Napoli C. Nitric oxide as a signaling molecule in the vascular system: an overview. J cardiovascular pharma. 1999; 34: 879-886. DOI: https://www.doi.org/10.1097/00005344-199912000-00016
Plumb Donald. Plum’s Veterinary Drug Handbook, Sixth Edition, Blackwell. 2008; 658– 659. Available at: https://www.amazon.com/
Donald-C-Plumb-Pharm-D-Veterinary/dp/B005H7TYM6
Brisinda M, Maria G, Bentivoglio AR, Cassetta E, Gui D, and Albanese A. A Comparison of Injections of Botulinum Toxin and Topical Nitroglycerin Ointment for the Treatment of Chronic Anal Fissure. N Engl J Med. 1999; 341: 65-69. DOI: https://www.doi.org/10.1056/NEJM199907083410201
Sanei B, Mahmoodieh M, and Masoudpour H. Comparison of topical glyceryl trinitrate with diltiazem ointment for the treatment of chronic anal fissure: a randomized clinical trial. Acta Chir Belg. 2009; 109: 727-730. DOI: https://www.doi.org/10.1080/00015458.2009.11680524
Ahmadinejad M, Khaksarhaddad M, Amirhasani A, Khazaeli P, AsadiShekari M, Ahmadipour M, and TahmasebyS. Compression of Efficacy of Topical Phenytoin, Honey and Combination of Honey with Triglyceride Tree Nitrate on Wound Healing in Rat. Indian J
Forensic Med and Toxicology. 2019; 13: 393-398. DOI: https://wwww.doi.org/10.5958/0973-9130.2019.00320.7
Heggers JP, Elzaim H, Garfield R, Goodheart R, Listengarten D, Zhao J, and Phillips LG. Effect of the combination of Aloe vera, nitroglycerin, and L-NAME on wound healing in the rat excisional model.
J Altern Complement Med. 1997; 3: 149-153. DOI: https://www.doi.org/10.1089/acm.1997.3.149
Ellabban MA, Fattah IOA, Kader GA and et al. The Effects of Sildenafil and/or Nitroglycerin on Random pattern Skin Flaps After
Nicotine Application in Rats. 2020; 10: 3212. DOI: https://www.doi.org/10.1038/s41598-020-60128-w
Dunn CL, Brodland DG and et al. A single postoperative application of nitroglycerin ointment does not increase survival of cutaneous
flaps and grafts. Dermatol surg. 2000; 26: 425-427. DOI: https://www.doi.org/10.1046/j.1524-4725.2000.99307.x
Cihan AO, Bicakci U, Tander B, Rizalar R, Kandemir B, Ariturk E, and Bernay F. Effects of intra peritoneal nitroglycerin on the strength and healing attitude of anastomosis of rat intestines with ischemia-reperfusion injury. Afr J Paediatr Surg. 2011; 8: 206-210. DOI: https://www.doi.org/10.4103/0189-6725.86064
Hwang DY, Yoon SG, Kim HS, Lee JK, and Kim KY. Effect of 0.2 percent glyceryl trinitrate ointment on wound healing after a hemorrhoidectomy: Results of a randomized, prospective, double-blind, placebo-controlled trial. Dis Colon Rectum. 2003; 46: 950-954. DOI: https://www.doi.org/10.1007/s10350-004-6692-0
Kim SW, Kim H, and Hyun M. Nitroglycerin Ointment Enhances the Wound Healing Effect of Adipose-Derived Stem Cells. Int J Stem Cells
Res. 2020; 4: 1-5. DOI: https://www.doi.org/10.1155/2019/2745640
Karanik H, Akturk R, Camlica H, and Asoqlu O. The effect of glyceryl trinitrate ointment on post hemorrhoidectomy pain and wound healing: Results of a randomized, double-blind, placebo-controlled study. Dis Colon Rectum. 2009; 52: 280-285. DOI: https://www.doi.org/10.1007/DCR.0b013e31819c98a7
Ratnasingham K, Uzzaman M, Andreani SM, Light D, and Patel B. Meta-analysis of the use of glyceryl trinitrate ointment after haemor-rhoidectomy as an analgesic and in promoting wound healing. Int J Surg. 2010; 8: 606-611. DOI: https://www.doi.org/10.1016/j.ijsu.2010.04.012
Cihan AO, Bicakci U, Tander B, Rizalar R, Kandemir B, Ariturk E, and Bernay F. Effects of intra peritoneal nitroglycerin on the strength and healing attitude of anastomosis of rat intestines with ischemia-reperfusion injury. Afr J Paediatr Surg. 2011; 8: 206-210. DOI: https://www.doi.org/10.4103/0189-6725.86064
Gündüz M, Sekmenli T, Uğurluoğlu C, and Çiftçi İ. The effects of nitroglycerin in the zone of stasis in a rat burn model. Ulus
Travma Acil Cerrahi Derg. 2020; 26: 171-177. DOI: https://www.doi.org/10.14744/tjtes.2019.00005
Hotkar MS, Avachat AM, Bhosale SS, Oswal YM. Preliminary investigation of topical nitroglycerin formulations containing natural wound healing agent in diabetes‐induced foot ulcer. International wound J. 2015; 12: 210-217. DOI: https://www.doi.org/10.1111/iwj.12084
Liu JW, Lin CC, Kiu KT, Wang CY, and Tam KW. Effect of glyceryl trinitrate ointment on pain control after hemorrhoidectomy: a meta-analysis of randomized controlled trials. World J surgery. 2016 ; 40: 215-224. DOI: https://www.doi.org/10.1007/s00268-015-3344-6
Flynn V, Chisholm C, and Grimwood R. Topical nitroglycerin: a promising treatment option for chondro dermatitis nodularis helicis. J Acad Dermatol. 2011; 65: 531-536. DOI: https://www.doi.org/10.1016/j.jaad.2010.06.012
Wang P, Gu L, Qin Z, and et al. Efficacy and safety of topical nitroglycerin in the prevention of mastectomy flap necrosis: a systematic review and meta-analysis. 2020; 10: 6753. DOI: https://www.doi.org/10.1038/s41598-020-63721-1
Challoumas D, Kirwan PD, Borysov D, and et al. Topical glyceryl trinitrate for the treatment of tendinopathies: a systematic review British J Sports Med. 2019; 53: 251-262. DOI: https: //www.doi.org10.1136/bjsports-2018-099552
Gambito ED, Gonzalez-Suarez CB, Oquiñena TI, and Agbayani RB. Evidence on the effectiveness of topical nitroglycerin in the treatment of tendinopathies: a systematic review and meta-analysis. Arch Phys Med Rehabil. 2010; 91: 1291-1305. DOI: https://www.doi.org/10.1016/j.apmr.2010.02.008.