In-vitro and In-vivo Evaluation of Cytotoxic Activity of Rutin Berberine Loaded Polycaprolactone Nanoparticles against EAC Cell Line

Abstract

Background and Purpose: Rutin, a flavonoid found in various species of plants like buck wheat, citrus fruits, asparagus, apples, berries and tea was found to possess various biological properties like antioxidant, anti-inflammatory, vasodilation, enzyme regulation, anticancer and neuroprotective effects. On the other hand, berberine is an alkaloid found in berberis species, Hydrastis canadensis and Coptic chinensis with wide range of properties like anti inflammatory, antioxidant and anti-microbial. Therefore, in this study our goal was to formulate rutin and berberine in combination loaded polycaprolactone nanoparticles and test its invitro and in vivo anticancer potential. Materials and methods: The nanoparticles were formulated using single emulsification method. Their physicochemical properties were gauged using SEM, FTIR, DLS, Drug content and invitro drug release. The antineoplastic activity of the drug loaded nanoparticles in vitro was assessed by MTT assay. The in vivo anti neoplastic activity was studied in female swiss albino mice using the EAC cell lines. Results: in vitro neotoxicity studies revealed that the Rutin berberine loaded PCL nanoparticles were cytotoxic in dose dependent manner as calculated by its percentage cell viability and also showed cellular damage and cell death features of apoptosis. The in vivo neotoxicity study reports as studied by the histopathological evaluation of liver revealed the reservation of hepatocytes with reduction in the necrotic cells and dysplastic hepatocytes showing pattern of recovery with the presence of Kupffer cells and in kidney showing no changes in glomerular dilation and tubular necrosis. Nuclei and nephron cells remained normal with no necrosis infiltration showing the drug loaded nanoparticles possess good anti tumor property. The haematological parameters as measured by Hb level, RBC, WBC, ESR And CRP and biochemical parameters as measured by total protein, creatinine, bilirubin, SGPT, SGOT, ALP, albumin, urea, uric acid, SOD, catalase, GSH, GPx,  GST and lipid peroxidase showed that the drug loaded PCL nanoparticles possessed good antineoplastic activity comparable with the standard drug Cisplatin. Conclusion: Rutin berberine loaded PCL nanoparticles are identified as an effective nanoparticulate system in treating the Ehrlich ascites carcinoma paving the way for further scientific research in humans.