Wound healing effects of Artemisia sieberi extract on the second degree burn in mice skin

Kaboutari Katadj J et al Journal of HerbMed Pharmacology, Volume 5, Number 2, April 2016 http://www.herbmedpharmacol.com 68 in the traditional and modern medicine as anti-malaria, anti-pyretic, analgesic, anti-oxidant, anti-inflammatory, anti-hepatitis, anti-ulcerogenic, anti-helminthic, antiamoebic and anti-hemorrhagic (9-13). Anti-ulcerogenic action of some sesquiterpene lactones of A. annua and ethanolic extract of its aerial parts had been shown (14,15). Histopathological and biochemical analysis of A. Anuua extract showed that it can hasten healing of experimental dermal injuries (16). Later accelerating effects of ethanolic extract of A. aucheri on wound healing and reducing the healing period was reported (17). Also the ability of A. sieberi extract to increase thickness of skin layers and collagen fibers have been reported (8). So due to anti-microbial, anti-inflammatory, anti-oxidant and antiulcerogenic activity of Artemisia species and its suitable effects on the histopathological, histomorphometric and biochemical parameters of the skin and wound healing, this study was conducted to evaluate the effects of A. sieberi extract on the healing of second degree burn. Materials and methods Plant extraction Aerial parts of A. sieberi were collected from Yazd province in the center of Iran, dried at room temperature, grounded and sieved and then Artemisinin extraction was done by using petroleum ether (8,11). Animals Ninety adult male mice (20-25 g) in three experiments were used. Animals were housed under standard laboratory conditions in accordance with the European community guidelines for laboratory animals (12 hours light/dark cycle, ambient temperature of 22 ± 1°C٫ 60% humidity) and under supervision of the Ethical Committee of the University, while chow pellet and water were freely available. After 1 week of acclimatization, they were divided accidentally into three groups (n = 30). Mice were anesthetized with an intra-peritoneal (i.p) injection of 40 mg/kg sodium thiopental (UVB Pharma, Czech Republic). The skin of dorsum was shaved and thoroughly depilated and thereinafter washed with povidone iodine. Burn was made by contact of a round 1 cm diameter metal plate (105°C, 5 seconds, without pressure) and then for prevention of shock, 1 ml of normal saline was injected i.p and each animal was kept in a separate cage. Group one was treated with SSD 1% ointment (King Pharmaceuticals, USA), group two was treated with A. sieberi extract but group three was selected as negative control and received no medication. Treatments were done every 12 hours for 21 days. Histopathological evaluation The burns were monitored for sign of infection and digital photographs were taken daily and after analysis by using ImageJ software, they were used for measurement of wound area and calculating healing percentage (18): [(area of the first dayarea of the second day)/ (area of the first day)]×100. For histopathological study, 5 mice of each group on the days 1, 3, 7, 14 and 21 post-treatments were chosen randomly and after euthanasia, a full thickness skin flap of the burn was taken and fixed. Tissue processing was done (Autotechnicon tissue processor Citadel 1000, Thermo Shandon, UK), specimens were stained with H&E and examined for tissue granulation, inflammation, re-epithelialization and collagen sediment (19,20). Hydroxyproline content of burn as the index of tissue collagen was measured (21). Statistical analysis Data were presented as mean±SD and analyzed by using SPSS version 16.0 (Chicago, IL, USA). Kruskal-Wallis and Dunn post hoc tests were used for data analysis (P ≤ 0.05). Results Healing percentage Healing percentage of A. sieberi and SSD was significantly higher than control on days 7&14 (P ≤ 0.05), while on day 7, the difference between A. sieberi and SSD was not significant (P > 0.05; Figure 1). Inflammation From day 1 to 21 inflammation score was decreasing, but only on days 3 & 7 in the A. sieberi and SSD groups it was significantly lower than control (P ≤ 0.05), while on the rest of days such a significant difference was not seen (P > 0.05; Figure 2). Granulation tissue On day 3 granulation tissue of A. sieberi and SSD was significantly higher than control (P ≤ 0.05), while there was no significant difference neither between A. sieberi and SSD on day 3, and nor between A. sieberi and SSD with control on other days (P > 0.05; Figure 3). Hydroxyproline Hydroxyproline content of burn as the index of collagen synthesis was measured on day 14. It was significantly