Evaluation of ghee based formulation for wound healing activity

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Abstract

Formulation containing neomycin and ghee was evaluated for wound-healing potential on different experimental models of wounds in rats. The rats were divided into six groups of group 1 as control, group 2 as treated with neomycin only, group 3 as treated only with ghee, group 4 treated with F-1 formulation containing ghee 40% and neomycin 0.5%, group 5 treated with F-2 formulation containing ghee 50% and neomycin 0.5% and group 6 treated with F-3 formulation containing ghee and ointment base in all two wound models, each group consisting of six rats. Wound contraction ability in excision wound model was measured at different time intervals and study was continued until wound is completely healed. Tensile strength was measured in 10-day-old incision wound and quantitative estimation of hydroxy proline content in the healed tissue was determined in 10-day-old excision wound. Histological studies were done on 10-day-old sections of regenerated tissue of incision wound. F-2 formulation containing ghee 50% and neomycin 0.5% showed statistically significant response, in terms of wound contracting ability, wound closure time, period of epithelization, tensile strength of the wound, regeneration of tissues at wound site when compared with the control group and these results were comparable to those of a reference neomycin ointment.

Introduction

Wound is defined as a loss or breaking of cellular and anatomic or functional continuity of living tissues (Ayello, 2005). Healing of wound is a biological process that is initiated by trauma and often terminated by scar formation (Rubin and Fabrex, 1996). The process of wound healing occurs in different phases such as coagulation, epithelization, granulation, collegenation and tissue remodeling (Fulzele et al., 2002). In India, there has been interest in the potential of natural products obtained from plants and animals for development of drugs with wound healing properties as taught in a popular form of Indian medicine known as Ayurveda (Biswas and Mukherjee, 2003). Natural products have been an important resource for the maintenance of life since ages; natural products are becoming increasingly important as alternative medicines. Wither directly as herbal drugs for the treatment of chronic diseases or as raw materials from which more or less complex chemical compounds with particular biological activity are isolated, the deeply held beliefs in the efficiency of natural products/herbals can and should be proven by scientific investigation bringing the tradition and experience in the ancient knowledge within the realm of science (Duke et al., 2002).

Side effects associated with allopathic drugs and synthetic compounds, has prompted research into herbal and natural products (Govindarajan et al., 2004). Panchgavya is a term used in Ayurveda to describe the five important bovine products viz. milk, curd, ghee, urine and dung. Several formulations based on each one of these components are reported in ayurvedic texts with medical claims. Individually as well, these components have been ascribed several therapeutic values as mentioned in Ashtangahriday Chikitsasthan, Kushta Chikista Shlok. 768. Several formulations based on panchgavya are described in ancient ayurvedic texts. Few scientific studies with regard to panchgavya are reported in modern literature/journals.

Ghee was selected, as it is widely used and easily available through out subcontinent and also cost effective. In the developing countries, ghee is used for cooking food items and it was used as the material for roasting the vegetables. It has also shown to be clinically effective in various clinical studies regarding its memory enhancing potential (Murray, 1996). Literature survey revealed that no scientific investigation has been made in regard to the wound healing activity of ghee and its combination with any antibiotic.

Ghee is an important panchgavya component. Ghee bears several medicinal claims and many of its combinations with herbs are reported in ancient texts. However, systematic chemical and clinical evaluation in this area is conspicuously lacking. One of the most widely circulated and well-known properties of ghee is its wound healing activity. The present work aims to systematically evaluate this claim with regard to ghee in combination with antibiotic. Ghee contains butyric acid, hexanoic and octanoic acids; these are removed as far as possible by distillation of mixed fatty acids. The volatile acids and mixture of glycerides other component acids of ghee are oleic acid, palmitic acid, myristic acid, stearic acid and characteristic proportion of butyric, caproic, caprylic and capric acids and minor amounts of lauric acid and arachidionic acid, lots of unsaturated and saturated fatty acids. Saturated fatty acids are butyric acid, caproic acid, caprylic acid, capric, lauric, myristic, palmitic, stearic and araeludic acids. The unsaturated fatty acids are deconic, dodeconic, tetradeconic, hexadeconic and octadedienoic acids (Murray and Watt, 1972).

Ghee is a product, which was received from Go-Vighyan Kendra, which is an organization works for products obtained from animals of bovine origin. The objective of this work is to verify the hypothesis that cows ghee by providing better tissue formation and neomycin (Katzung, 1995) by providing protection against invasion would prove a superior treatment for healing wounds if given concomitantly preferably in one single formulation and to examine whether such formulation satisfies the normal criteria for a formulation to be accepted as a medicinal agent.

Therefore the aim of the present study was to evaluate the wound healing property of the ghee and neomycin as active constituents in simple ointment base using various experimental wound-healing models (Mukerjee et al., 2000).

Section snippets

Materials

Neomycin sulphate was obtained as the gift sample from Unichem Laboratories, Mumbai. Ghee was obtained as the gift sample from Go-Vighyan Anusandhan Kendra, Nagpur. Sodiumbisulphite, glyceryl monostearate, cetylalchol, propyl paraben and methyl paraben were obtained from Sigma–Aldrich Chemie, Germany. All the solvents used for titrations for estimating free fatty acid level and peroxide level are of analytical grade obtained from Spetrochem Pvt. Ltd., Mumbai.

Preparation of formulation

Ointments were prepared by

Results

The process of wound healing has two components, first is formation of new tissue and other is protections from microbial invasion during the healing process. Cow's ghee promotes the former while neomycin offers the later. The logical manner of utilization of these two properties would be to formulate these two substances in one single formulation.

With this aim in mind a couple of formulations were attempted in which cow's ghee and neomycin was combined in single formulation. Neomycin was

Discussion

The aim of this present work is to verify the hypothesis that cows ghee by providing better tissue formation and neomycin (Katzung, 1995) by providing protection against microbial invasion would prove a superior treatment for healing wounds if given concomitantly preferably in one formulation. Somewhat consequent to this objective it became necessary to develop a formulation containing these two active medicaments namely ghee and neomycin. It became consequential to examine whether such a

Conclusion

It is possible to prepare a formulation containing ghee with neomycin, the formulation was physically and chemically stable for at least 45 days at 40 °C, the findings of panel evaluation of different formulations F-1, F-2 and F-3. F-2 was superior in texture and spreadability to remaining formulations, the formulations are also chemically stable and there was no significant change in the free fatty acid value and peroxide value. The findings meet with the standards of AOCS and Bureau of Indian

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