Oxidative stress is considered to be the leading cause of many chronic and degenerative diseases leading to death and disability. Antioxidant therapy could be efficient in preventing oxidative stress-induced damages in cells. In the present study the efficacy of mangiferin-chitosan nanoparticles (MCNs) as a therapeutic agent against the oxidative stress, since mangiferin (MGF), a polyphenol protects and attenuates oxidative stress against various diseases. However, the bioactivity of MGF does not result in vivo biological effect owing to its low bioavailability and poor solubility. Mangiferin nanoparticles (NPs) were developed to improve the solubility and bioavailability of mangiferin. Nanoparticles had significant free radical scavenging activity in the cell-free system and had a synergistic action with phase II antioxidant enzymes such as catalases and peroxidases. A nephropathic system was developed to investigate the attenuative role of mangiferin-chitosan nanoparticles against induced oxidative stress, on normal kidney epithelial (NKE) cells. Pre-treatment with the nanoparticles exclusively prevented the induction of cytotoxicity induced by NaF and maintained the level of intracellular antioxidant enzyme in the cells. Nanoparticles had significant lipid peroxidation and protein oxidation inhibition activities. These nanoparticles can be used in food and pharmaceutical industries as a therapeutic agent to prevent the oxidative stress-induced health disorders.