Histopathology and Histomorphological Study of Wound Healing Using Clove Extract Nanofibers (Eugenol) Compared to Zinc Oxide Nanofibers on the Skin of Rats

Document Type : Original Articles

Authors

1 Department of Clinical Science, Faculty of Specialized Veterinary Science, Science and Research Branch, Islamic Azad University, Tehran, Iran

2 Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

3 Universal Scientific Education and Research Network, Tabriz, Iran

4 Department of Pathology, Science and Research Branch, Islamic Azad University, Tehran, Iran

Abstract

In order to evaluate the healing effect of eugenol and other nanofibers, 100 male Wistar rats (200±10 g) were used with 14-15 weeks of age in this study. All of the male rats were transferred in the standard cages under controlled exposure conditions in a 12:12 h light/dark cycle with a constant temperature about 22±2 oC. In addition, the male rats were fed with pellets. Firstly, anesthesia process was performed by 2% xylazine hydrochloride (10mg/Kg/IP) and 10% ketamine hydrochloride (100mg/Kg/IP), and then the rats were placed on the operating table. Then the dorsal surfaces of the rats’ skin to ileum were scrubbed and prepared as the next step. A circular wound (with a 7 mm diameter) was created by a 7 mm sterile biopsy punch. All 100 rats were divided into four groups (n=25) randomly named as control, nano zinc oxide (ZnO), eugenol nanofibers, and polycaprolactone groups. After that, they were divided into five groups regarding the wound closure rate in days 3, 5, 7, 14, and 21. Then, the wound dressings were placed on the wounds and renewed every 24 h. At the end of days 3, 5, 7, 14, and 21, the relevant tests, such as histopathology, were conducted by removing the tissue volume using a biopsy punch, and then decapitation process was performed on the rats. It was obvious that eugenol nanofiber showed the best granulation tissue by the production of collagen. Further studies are being performed on wound healing by eugenol nanofiber. 

Keywords

Main Subjects


Article Title [French]

Étude Histopathologique et Histomorphologique de la Guérison Des Plaies en Utilisant des Nanofibres d'Extrait de Clou de Girofle (Eugénol) Comparées aux Nanofibres d'Oxyde de Zinc sur la Peau de Rats

Abstract [French]

Dans cette étude, 100 rats Wistar mâles (200 ± 10 g) ont été utilisés à l'âge de 14-15 semaines, afin d'évaluer l'effet cicatrisant de l'eugénol et d'autres nanofibres, Tous les rats ont été transférés dans des cages standards dans des conditions d'exposition contrôlées comprenant un cycle lumière / obscurité de 12:12 et une température constante d'environ 22 ± 2 oC. De plus, les rats ont été nourris avec des pellets. En premier lieu et juste avant de placer les rats sur la table d'opération, une anesthésie a été effectuée utilisant 2% de chlorhydrate de xylazine (10 mg / kg / IP) et 10% de chlorhydrate de kétamine (100 mg / kg / IP). Ensuite, la surface dorsale de la peau jusqu’à l’iléon des rats a été nettoyée et préparée pour l’étape suivante. Une plaie circulaire (d'un diamètre de 7 mm) a été induite par le biais d’un perforateur de biopsie stérile de 7 mm. Les 100 rats ont été ensuite divisés en quatre groupes (n = 25), considérés de manière aléatoire comme les groupes témoins, nano-oxyde de zinc (ZnO), nanofibres d'eugénol et polycaprolactone. Les rats composant chaque groupe ont été ensuite également distribués dans cinq sous-groupes afin d’évaluer le taux de fermeture des plaies aux jours 3, 5, 7, 14 et 21. Des pansements ont été placés sur les plaies et renouvelés toutes les 24 heures. Au terme des jours 3, 5, 7, 14 et 21, les tests d'histopathologie ont été réalisés en retirant une portion du tissu à l'aide d'un poinçon pour biopsie avant de procéder à la décapitation des rats. Il était évident que la nanofibre d'eugénol présentait le meilleur tissu de granulation grâce à la production de collagène. D'autres études sont en cours sur la cicatrisation des plaies à l'aide de nanofibres d'eugénol.

Keywords [French]

  • Nanofibres
  • Eugénol
  • Oxyde de zinc
  • Peau
  • rat
  • Cicatrisation des plaies
Alam, P., Ansari, M.J., Anwer, M.K., Raish, M., Kamal, Y.K.T., Shakeel, F., 2016. Wound healing effects of nanoemulsion containing clove essential oil. Artif Cells Nanomed Biotechnol 45, 591-597.
Briozzo, J., Nunez, L., Chirife, J., Herszage, L., D'Aquino, M., 1989. Antimicrobial activity of clove oil dispersed in a concentrated sugar solution. J Appl Bacteriol 66, 69-75.
Chaieb, K., Zmantar, T., Ksouri, R., Hajlaoui, H., Mahdouani, K., Abdelly, C., et al., 2007. Antioxidant properties of the essential oil of Eugenia caryophyllata and its antifungal activity against a large number of clinical Candida species. Mycoses 50, 403-406.
Charernsriwilaiwat, N., Opanasopit, P., Rojanarata, T., Ngawhirunpat, T., 2012. Lysozyme-loaded, electrospun chitosan-based nanofiber mats for wound healing. Int J Pharm 427, 379-384.
Cushnie, T.P., Lamb, A.J., 2011. Recent advances in understanding the antibacterial properties of flavonoids. Int J Antimicrob Agents 38, 99-107.
Diegelmann, R.F., Evans, M.C., 2004. Wound healing: an overview of acute, fibrotic and delayed healing. Front Biosci 9, 283-289.
Flecknell, P.A., 1993. Anaesthesia of animals for biomedical research. Br J Anaesth 71, 885-894.
Griga, T., Tromm, A., Spranger, J., May, B., 1998. Increased serum levels of vascular endothelial growth factor in patients with inflammatory bowel disease. Scand J Gastroenterol 33, 504-508.
Hemaiswarya, S., Doble, M., 2009. Synergistic interaction of eugenol with antibiotics against Gram negative bacteria. Phytomedicine 16, 997-1005.
Kong, X., Liu, X., Li, J., Yang, Y., 2014. Advances in pharmacological research of eugenol. Curr Opin Complement Alternat Med 1, 8-11.
Kurian, R., Arulmozhi, D., Veeranjaneyulu, A., Bodhankar, S., 2006. Effect of eugenol on animal models of nociception. Indian J Pharmacol 38, 341.
Nangle, M.R., Gibson, T.M., Cotter, M.A., Cameron, N.E., 2006. Effects of eugenol on nerve and vascular dysfunction in streptozotocin-diabetic rats. Planta Med 72, 494-500.
Nayak, B., Pinto Pereira L.M., 2006. Catharanthus roseus flower extract has wound-healing activity in Sprague Dawley rats. BMC Complem Altern M 6, 41.
Nissen, N.N., Polverini, P.J., Koch, A.E., Volin, M.V., Gamelli, R.L., DiPietro, L.A., 1998. Vascular endothelial growth factor mediates angiogenic activity during the proliferative phase of wound healing. Am J Pathol 152, 1445-1452.
Núñez, L., D'Aquino, M., Chirife, J., 2001. Antifungal properties of clove oil (Eugenia caryophylata) in sugar solution. Braz J Microbiol 32, 123-126.
Phillips, C., 2008. The welfare of animals: the silent majority. Springer Science & Business Media.
Prabhakaran, M.P., Venugopal, J.R., Ramakrishna, S., 2009. Mesenchymal stem cell differentiation to neuronal cells on electrospun nanofibrous substrates for nerve tissue engineering. Biomaterials 30, 4996-5003.
Singer, A.J., Clark, R.A., 1999. Cutaneous wound healing. N Engl J Med 341, 738-746.
Suckow, M.A., Stevens, K.A., Wilson, R.P., 2012. The laboratory rabbit, guinea pig, hamster, and other rodents. First ed. Academic Press, London.
Suganya, S., Senthil Ram, T., Lakshmi, B., Giridev, V., 2011. Herbal drug incorporated antibacterial nanofibrous mat fabricated by electrospinning: an excellent matrix for wound dressings. J Appl Polym Sci 121, 2893-2899.
Taher, Y.A., Samud, A.M., El-Taher, F.E., ben-Hussin, G., Elmezogi, J.S., Al-Mehdawi, B.F., et al., 2015. Experimental evaluation of anti-inflammatory, antinociceptive and antipyretic activities of clove oil in mice. Libyan J Med 10, 28685.
Wellington, D., Mikaelian, I., Singer, L., 2013. Comparison of ketamine–xylazine and ketamine–dexmedetomidine anesthesia and intraperitoneal tolerance in rats. J Am Assoc Lab Anim 52, 481-487.
Yan, S., Xiaoqiang, L., Lianjiang, T., Chen, H., Xiumei, M., 2009. Poly (l-lactide-co-ɛ-caprolactone) electrospun nanofibers for encapsulating and sustained releasing proteins. Polymer 50, 4212-4219.
Yazdanpanah, L., Mohamadi, N., 2014. Antifungal activity of clove essential oil from Syzygium aromaticum on Paecilomyces variotii agent of pistachio dieback. J Biodivers Environ Sci 4, 42-45.
Young, S.R., Dyson, M., 1990. Effect of therapeutic ultrasound on the healing of full-thickness excised skin lesions. Ultrasonics 28, 175-180.