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GC-MS Analysis and Wound Healing Efficiency of Ethanolic Leaves Extract of Cassia occidentalis and Pithecellobium dulce in Wistar Albino Rats: Comparative Study

Aim: To identify and compare the bioactive compounds in the ethanolic leaves extracts of Cassia occidentalis and Pithecellobium dulce and to evaluate the wound healing efficiency in Wistar Albino rats. 
Study Design: The leaves ethanolic extracts was analysed by GC-MS and the extract was prepared in the form of a cream by ethanolic leaves extracts of C. occidentalis and P. dulce at 5% (w/v), 10% (w/v), and also in combination, a simple ointment base was developed with a composition of (1:1) Topical application of 5% (w/v) and 10% (w/v) was utilised in excision wound models. For excision wound models, the treatment duration was ten days. The day on which the wound was inflicted was designated as day '0'. Wound healing Activity: Excision wound Model: The animals were randomly separated into eight groups of six rats each: Group I: Control.; Group II: Standard group, treated with Framycetin sulfate cream (Soframycin, Aventis);. Group III: Treated with ethanolic extract of C. occidentalis (ELCO) (5% w/v); Group IV: Treated with ethanolic extract of C. occidentalis (ELCO) (10% w/v);Group V: Treated with ethanolic extract of P. dulce (ELPD)(5% w/v); Group VI: Treated with ethanolic extract of P. dulce (ELPD) (10 % w/v);Group VII: Treated with ethanolic extract of C. occidentalis and P. dulce (ELCO & ELPD 1:1) (5% w/v); Group VIII: Treated with ethanolic extract of C. occidentalis and P. dulce (ELCO & ELPD 1:1) (10 % w/v) till complete epithelization. Next dead space wound model and histology was studied. 
Place and Duration of Study: The GC-MS was carried out at Lab in Chennai. The extraction procedures were done at Department of Microbiology, Acharya Nagarjuna University, Guntur and treatment of wound healing activities were conducted at Ratnam Institute of Pharmacy in Nellore, Andhra Pradesh, India, and housed in the Department of Pharmacology between October to January 2016. 
Methodology: To study bioactive compounds, GC-MS was adopted, for wound healing activity: Excision wound Model, Dead space wound model and histology procedures was applied. 
Results:  In the current study, ethanol leaves extract (EL) of Cassia occidentalis and Pithecellobium dulce were compared using GC-MS and their wound healing efficacy in wistar rats was examined. The GC-MS analysis of EL from both plants revealed 14-16 distinct bioactive phytochemical components with varying molecular weights and retention duration (RT). Excision and dead space wound models were utilised to assess the wound healing activities of EL extracts on rats. Wound concentration, full epithelialzation time, granulation, tissue weight, and hydroxyproline content were used to measure healing. In the excision wound model, the standard group (Framycetin sulphate cream) and group-VII (10% w/v; 1:1) combination EL treatment exhibited 98.5 ± 0.54 % and 98.4 ± 0.46 % wound healing activity, respectively. When compared to the control, the granulation tissue weight and hydroxyproline content in the dead space wound rose considerably. Histological examination revealed fewer inflammatory cells and more collagen, indicating a role in accelerating wound healing activity. 
Conclusion: The results of our investigation indicate unequivocally that ethanolic leaf extracts of these plant species are effective at encouraging wound healing. The 10% (ELCO+ELPD) tropical treatment drastically reduced the wound as compared to standard and also increased granulation and hydroxyproline content. However, it requires more clinical examination before being considered for wound therapy.