ACTIVITIES OF TRIPHALA TOWARDS PROMOTING COLLAGEN SYNTHESIS AT WOUND SITE AND INHIBITING METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS AND ITS ENZYMES

Problem Statement: Infection is a major problem in the management of wounds. Despite the use of synthetic antimicrobial agents, drug resistance and toxicity hinder the activity of these antimicrobial agents; thereby increase the chances of infection. The microbial enzymes produced by wound pathogens degrade the extra cellular matrix at wound site. The present study investigates the influence of Triphala on changes in collagen characteristics during the healing process of an infected cutaneous wound in Albino Wister rats and also how the antimicrobial activity and enzyme inhibition activity of triphala used in wound healing studies. Approach: A methanol extract of triphala was prepared and its antimicrobial activity was tested against twenty clinical MRSA strains. The activity of triphala extract against serine protease and metalloprotease was studied by Zymography. Enzymatic activities were detected as clear bands of casein/gelatin lysis against a dark background. To measure the relative enzyme levels, clear zones were scanned and the percentage of inhibition was analyzed by Gel Documentation systems. The inhibition of enzymes by Triphala has been expressed in percentage and expressed as mean ± SD of ten experiments. Male Wister albino rats of weights ranging between 150g and 200g were used for in vivo wound healing study. Granulated tissues were collected on the 4th, 8th, 12th and 16th days for the estimation of different types of collagen present in the granulated tissue and also for histological studies. Statistical Analysis: All results are expressed as a mean  S.D and the results were compared statistically by a student’s independent t- test using SPPS software. A statistically significant p value of less than 0.05 was considered. Result: The 182mm clear zone in disc diffusion assay and minimal inhibitory concentration (MIC) of 7.8125mg/ml against MRSA (as well as methicilin susceptible Staphylococcus aureus) control strains clearly showed the antibacterial activity of Triphala. Zymography analysis exhibited greater reduction in serine protease and metalloprotease activity at 1500g/ml. The wound tissues that were removed on the 4th, 8th, 12th and 16th day (post-wound), were used to analyze the biochemical and pathological changes on the injured tissue. Triphala increased cellular proliferation and collagen synthesis at the wound site, as evidenced by increase in type III collagen content of wound tissues. The Masson’s Trichrome staining of granulated tissue confirmed that the treated subset of tissues had well-formed epithilization with well stretched bundles of collagen when compared to that of an open wound group (untreated). Better maturation and cross linking of collagen was observed in those rats which were treated with Triphala. Conclusion: By virtue of the inhibitory effect of Triphalaon different MRSA strains and their enzymes such as serine protease and metalloprotease, it could be, potentially used as a new therapeutic agent for MRSA infected dermal wounds. The results hence highlighted the beneficial effects of the topical application of Triphala in the acceleration of wound healing and its effect on collagen synthesis.