Anthocyanins-loaded eudragit® l100 nanoparticles: In vitro cytotoxic and genotoxic analysis

Abstract

Anthocyanins are flavonoids compounds that have a great therapeutic potential acting as antioxidants, anti-inflammatory and anti-carcinogenic agents. They are consumed through food, pharmaceutical and cosmetics industries as a natural alternative to synthetic dyes. However, anthocyanins are unstable depending on pH, temperature, light and oxygen variations, which led to the production of its encapsulation in nanoparticles as an alternative to increase its stability and protect them from chemical degradation for human consumption. Due to the fact that many of the beneficial properties of nanoparticles could also be harmful to human health, and the need of prior toxicological evaluation of any medicinal product, this research aims to evaluate the cytotoxic and genotoxic effects of anthocyanins-loaded Eudragit® L100 nanoparticles (AN). A range of AN doses were tested with a preliminary MTT assay, which allowed selecting five concentrations for comet and micronucleus assays: 0.2, 2, 20, 125 and 250 µg/mL, respectively, on human peripheral blood mononuclear cells (PBMC) (cells without liver metabolizing enzymes) and human hepatoma cell line HepG2 (with liver metabolizing enzymes). Results showed absence of cytotoxic effect in MTT test on both cell types after 24 h of exposure. However, the nuclear division index in PBMC cells assessed by micronucleus test indicates a decrease in the cell division, after 28 h of exposure. Genotoxicity analysis showed that AN did not produce significant genotoxic effects detected in comet assay, in either cell type. However, the micronucleus test on HepG2 cells showed that at concentration of 2 µg/mL and higher, AN produced clastogenic/aneugenic effects. Under our experimental conditions and limitations, the observed cytotoxic and genotoxic effects, not previously reported in the literature, may be due to anthocyanin metabolization and related to its concentrations and time of exposure.