Elucidating the physicochemical characteristic and cytotoxic activity of chitosan based nanoparticles with gynura procumbens extract as a new modular treatment against canine mammary gland tumor

An abstract of the research paper presented to the Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, in partial requirement on the course DVT55204 – Research Project.
Abstract: Canine mammary gland tumours (CMGT) are common in intact bitches. Although surgical excision and chemotherapy are the main therapies for CMGT, but these approaches frequently result in high recurrence rates and adverse effects, including cardiotoxicity and nausea. Chitosan, a natural polysaccharide found in crustaceans, and (GP), a plant that has been traditionally used to treat a variety of ailments, have been shown to have potential as part of cancer treatment with fewer side effects. However, their used in veterinary medicine, particularly in treating CMGT is still unestablished. Therefore, this study aims to analyse the physicochemical characteristics of chitosan nanoparticle formulation, assess in vitro cytotoxic effects of chitosan nanoparticle and GP extract on mammary tumor cells, and identify phytochemical constituents in GP extract. Investigations encompassed physicochemical analysis of the chitosan formulation using dynamic light scattering (DLS) zetasizer, evaluation of in vitro cytotoxic effects using 4T1 cell lines, and identification of phytochemical constituents in GP extract qualitative and quantitatively. The cytotoxic activity of Chitosan, Chitosan-GP and GP extract was assessed using MTT assay against 4T1 cell line for 72 hours. The cell scratch migration assay was performed to investigate the migration inhibition potential of these substance. The result demonstrated Chitosan nanoparticles, produced using the ionic gelation technique, showed a size of 340 nm, a polydispersity index of 0.3, and zeta potential of 14 mV. The GP extract revealed the presence of alkaloids and tannins in GP extract, with significant total phenolic content (79.808 mg of GAE/g) and total flavonoid content (61.621 mg of QE/g), respectively. Cytotoxicity assessments demonstrated highly active cytotoxic activity of Chitosan and chitosan-GP extract against 4T1 cells. However, GP extract was shown weak cytotoxic against 4T1 cells. The Cell Migration Assay revealed inhibitory effects with treatment of Chitosan and Chitosan -GP extract shown the most significant inhibition followed by the GP extract. Results from this experiment indicate that GP extract, Chitosan, and Chitosan-GP exhibit cytotoxic activity against 4T1 cells, suggesting a potential modular treatment for CMGT. Overall these findings would create potential of new paradigm in the search of an effective therapeutic strategies against CMGT in veterinary oncology.
Keywords: Canine mammary gland tumour (CMGT), Chitosan nanoparticles, Physicochemical characteristics, Cytotoxic activity, Gynura procumbens extract