Original Article BIOMECHANICAL, BIOCHEMICAL AND HISTOLOGICAL EVIDENCES FOR WOUND HEALING PROPERTIES OF INDIAN TRADITIONAL MEDICINES SUSHMA R. KOTIAN 1 , K. SREEDHARA R. PAI 2 , J. K. NAYAK 3 , HEMALATHA BANGERA 1 , KEERTHANA PRASAD 4 , KUMAR M. R. BHAT 1 1 Department of Anatomy, Kasturba Medical College, 2 Department of Pharmacology, Manipal College of Pharmaceutical Sciences, Manipal University, Manipal, 3 Sri Dharmasthala Manjunatheshwara Ayurvedic College, Udyavara, Udupi, 4 Department of Information Sciences, Manipal School of Information Sciences, Manipal University, Manipal, Karnataka, India Email: kumar.mr@manipal.edu Received: 19 Jul 2015 Revised and Accepted: 22 Sep 2015 ABSTRACT Objective: Ayurveda, India's traditional medicinal system is a rich source of natural remedies, frequently used as home and folk medicine in wound healing due to easy availability and affordability. Honey, Ghee and roots of Glycyrrhiza glabra are effectively used in Ayurveda for treating wounds of various types. Nerium indicum (a folk medicine) is also a known healing agent. Even though the known end result of these medications is faster wound healing, the mechanism of actions at tissue level, changes in the micro-environment of the wound and quantification of the rate of healing is not explored and documented using modern scientific methods. Methods: Healthy Wistar rats were used for incision wound model. Wounds were inflicted and the treatment plan was followed with regular topical application of test materials. The nature of healing was observed regularly and photographed. At different interval of the treatment plan-biomechanical, biochemical and histological studies were carried out. An attempt was also made to quantify the microscopic changes at the wound site. Results: Faster healing was observed in all the animals treated with test materials. This was indicated by alterations in the nature of epithelisation, inflammatory changes, fibroblast recruitment and activity, fibrous composition and arrangement at the wound site in comparison with untreated group. Conclusion: The present study is useful in exploring the mechanism of action of these traditional Indian medicinal systems–Ayurveda and folk medicine and thereby provides scientific evidences for the same. Keywords: Wound healing, Honey, Ghee, Glycyrrhiza glabra, Nerium indicum. INTRODUCTION Wound is defined as a loss or breaking of cellular and anatomic or functional continuity of living tissues which may not only be influenced by the external factors, but also as a complication of some disease processes [1] and other internal factors. Healing of the wound is a biological process that is initiated by trauma and often terminated by scar formation. Wound healing is classically divided into homeostasis, inflammation, proliferation, and remodelling [2]. The process of wound healing occurs in different phases such as coagulation, epithelisation, granulation, collagenation and tissue remodelling [3]. In spite of advancement in the modern medicine in wound management, active wound healing measures are still in the formative stage. In developing countries, there are additional disadvantages in wound management due to lack of adequate health care facilities, and inaccessibility to health centers. Therefore, the alternative medicine such as Ayurveda and folk medicinal systems are preferred as home remedy for treatment and management of the wound. Few such alternative treatment strategies are the use of Honey, Ghee and Yastimadhu (Glycyrrhiza glabra-GG) (Sushrutha samhitha) and a folk medicine Nerium indicum (NI), which is effectively practiced to treat various types of wounds since many years in India. Several studies show the role of individual application of Ghee [4] or Honey [5, 6] in wound management. GG is well known for its wound healing properties in Ayurveda. Several studies have proved the wound healing, anti-ulcer, anti-inflammatory and skin regeneration activity of GG [7]. NI is another herb used to treat wounds in the folk medicine whose efficacy however is not documented by clinical research [8]. Most of the studies addressed only the end results such as faster healing, closure of wound. The rate of re-epithelisation, nature of remodelling, and the quantification of healing at the wound site is seldom discussed. Further, there are no studies to show the combination effect of honey, Ghee, GG and NI on wound healing with emphasis on changes in the microenvironment of the wound site. The present study is therefore designed to scientifically evaluate the micro-mechanisms of wound healing activities of topical application of Honey, Ghee, Extracts of GG and NI singly or in combination which may give an insight into wound healing and may lead to the development of new strategies to manage the wound more effectively. MATERIALS AND METHODS Procurement and preparation of test material Cow’s ghee, Glycyrrhiza roots were obtained from Sri Dharmasthala Manjunatheshwara (SDM) Ayurvedic pharmacy, Udyavara, Udupi. Unprocessed honey in its raw form was procured from SDM Ayurvedic pharmacy. NI was procured in the month of December from its natural habitat in Udupi, and was identified by Dr. K. Gopal krishna Bhat, Professor of Botany (Rtd), Poornaprajna College, Udupi, Karnataka, India. A voucher specimen has been preserved in the Dept. of Pharmacognosy, Manipal College of Pharmaceutical Sciences, Manipal (PP no. 603). Preparation of aqueous extract OF GG and NI The roots of GG and leaves of NI after procurement were shade dried for seven days and then powdered. Aqueous extraction was carried out by hot maceration method [9]. The powdered plant materials (200g each) were dissolved in 1500 ml of distilled water and decoction was prepared at 75-80 degrees Celsius. Decoction was then cooled and filtered. Finally the filtrate was evaporated to dryness using lyophilizer. Phytochemical screening The aqueous extracts of GG and NI were subjected to preliminary phytochemical screening by using the standard protocol to detect the presence or absence of active constituents [10]. International Journal of Pharmacy and Pharmaceutical Sciences ISSN- 0975-1491 Vol 7, Issue 11, 2015 Innovare Academic Sciences