In Vivo Evaluation of Antirrhinum majus’ Wound-Healing Activity

Scientia Pharmaceutica Article In Vivo Evaluation of Antirrhinum majus’ Wound-Healing Activity Fadi G. Saqallah 1 , Wafaa M. Hamed 1 and Wamidh H. Talib 2, * 1 2 * Department of Pharmaceutical Chemistry and Pharmacognosy, Applied Science Private University, 11931 Amman, Jordan; (F.G.S.); (W.M.H.) Department of Clinical Pharmacy and Therapeutics, Applied Science Private University, 11931 Amman, Jordan Correspondence: ; Tel.: +962-799-840987 Received: 6 August 2018; Accepted: 25 September 2018; Published: 7 October 2018



 Abstract: Mediterranean-native perennial plant Antirrhinum majus was scrutinized in this study for its antioxidant activity and its total phenolic content in order to test for the plant’s wound-healing capability. The traditional uses of this plant to treat gum scurvy, various tumors, ulcers, and hemorrhoids were the main idea behind this study. Leaves and flowers of the A. majus were extracted by maceration. Pilot qualitative phytochemical tests were made to check the presence of various secondary metabolites. Quantitatively, the flowers’ macerate indicated superlative results regarding antioxidant activity and total phenolic content. However, the in vivo wound-healing capability study was made using 30 Wistar strain albino rats. This innovative part of the study revealed that the healing power of the flowers’ extract ointment (5% w/w) was superior compared to the leaves’ extract (5% w/w) and the positive-control ointments (MEBO) (1.5% w/w) (p ≤ 0.001). This activity was assessed by visual examination, wound-length measurement, and estimation of hydroxyproline content. Antirrhinum majus is a promising plant to be considered for wound healing. However, further testing (including histological examination and high-performance liquid chromatography (HPLC) analysis) is necessary to understand more about its mechanisms of action. Keywords: Antirrhinum majus; maceration extraction; antioxidant activity; total phenolic content; wound healing 1. Introduction According to traditional European medicine, based on the Hippocratic theory, the human body consists of a set of elements—humors—that share nature’s principal properties with the human body. They play a major role in physical and mental health, and can be influenced by individual diet, lifestyle, geography, and seasons. They neither are true fluids nor can be isolated. These humors are Sanguis, which stands for the red blood that emerges from lesions and wounds, Phlegm for watery secretions, Choler for the yellow bile in vomit and pus, and Melancholer, perhaps for the black bile that is clotted blood in veins and stool [1]. Humanity sought balance between these humors in various ways, which is where herbal medicine played a major role in keeping that balance as ancient populations and traditions started the gathering of natural products and plants and testing their healing effects [1,2]. Generally, wounds are classified according to their healing time into acute and chronic wounds. Acute wounds resolve within less than six weeks and are a result of a physical trauma or surgery, whereas chronic wounds do not resolve within six weeks, and their causes vary according to genesis, depth, care received, and, most importantly, the blood supply to the wounded tissue [3]. Normal tissue response to injury involves three basic phases: inflammation, new tissue formation, and remodeling. Inflammation starts with the coagulation cascade in order to prevent any excess loss of blood, get rid Sci. Pharm. 2018, 86, 45; doi:10.3390/scipharm86040045 www.mdpi.com/journal/scipharm Sci. Pharm. 2018, 86, 45 2 of 16 of dead tissues, and neutralize any possible invading pathogen. A platelet plug is formed. Fibrin and neutrophils are also involved. The second phase, new tissue formation, starts shortly after the completion of the first one, where migration of keratinocytes and angiogenesis take place and replace the granulation tissue and the fibrin matrix. The remodeling phase takes place throughout the downregulation of the inflammatory cascade. Remodeling includes apoptosis of the cells involved in the repair process from the previous phase, and the replacement of dead cells by collagen and other extracellular proteins [4]. The use of plants for the stimulation of wound healing has existed since ancient empires. Aloe vera, for example, is one of the most popular plants and was used as early as 1500 BC by the ancient Egyptians for the treatment of wounds. The leaves’ gel is thought to assist in the wound-healing process by keeping the tissue moist, promoting epithelialisation and tissue oxygenation, encouraging collagen and hyaluronic acid synthesis and deposition, and ceasing inflammation [4]. Various bioactive plants’ metabolites are responsible for wound-healing activity. Polyphenols are thought of being the key components in wound healing. Polyphenols generally demonstrate antimicrobial activity, antioxidant properties that enhance cell proliferation, and collagen production [5]. Other components like alkaloids, saponins, sulphur-containing compounds, fatty oils, phytosterols, proteins, resins, and many others were also evaluated for their contribution in the wound-healing process [4]. The Antirrhinum genus was originally enlisted under the Scrophulariaceae family tree until 1995 when a study [6] came to reclassify this genus under the Plantaginaceae family [7]. Antirrhinum majus is a Mediterranean-native perennial plant, which was used historically for inheritance and mutation studies by Charles Darwin thanks to its ease of cultivation, diploid inheritance, and variation in morphology and flowers’ colors [8,9]. A. majus is used only as a decorative plant for mass display in Middle Eastern countries [10]. In some traditions, the whole-plant decoction was used for the treatment of liver ailments and to resolve watery eyes [11,12]. It was also used as a treatment for gum scurvy and various tumors, ulcers, and hemorrhoids [9]. Analyzing A. majus throughout multiple studies showed the presence of multiple active metabolites. For example, aurones, minor flavonoids, were found existing in the flowers’ petals only, giving a bright yellow color and are likely produced from chalcones [13]. Aurones play a major role acting as antioxidant and antiproliferative agents against reactive oxygen species and reactive nitrogen species that contribute to aging, mutagenesis, carcinogenesis, and deoxyribonucleic acid damages by inhibiting oxidase enzymes [14]. On the other hand, multiple aucubin-like iridoids were found to be produced by A. majus as a defence mechanism against herbivores. These iridoids were defined as antirrhinoside, antirrhide, 5-glucosyl-antirrhinoside, linarioside, and chaenorrhinoside [15,16]. Polyunsaturated fatty acids and monounsaturated fatty acids were found to contribute in high percentages in the seed oil of A. majus. Both can be considered of great nutritional applications regarding card (...truncated)