Folic acid embedded in argentinian chitosan: antioxidant capacity and cytotoxic effect in Caco-2

CORFIELD ROCÍO; SCHEBOR CAROLINA C.; DE MATTEO REGINA; PEREZ OSCAR E.

CABA

Simposio; Exploring the frontiers of Chemistry; 2019

Universidad de Buenos Aires

Chitosan (CS) is a derivative of chitin obtained by a deacetylation process. It is a polymer widely used as an encapsulating agent for the reason that it is non-toxic, biodegradable and biocompatible (Zhang et al., 2019). The cost of purified polymer is high and the viability in industrial application is low, so the possibility of using CS with high molecular weight produced in Argentina is interesting. Folic acid (FA) is an essential vitamin in women of reproductive age as it is associated with the decrease of neural tube defects in fetus. This vitamin is incorporated through diet and is sensitive to environmental factors associated with food processing (Wusigale et al., 2017), so the nano-encapsulation methodology is evaluated in order to protect it. The Caco-2 cell line is a continuous line of human epithelial colorectal adenocarcinoma cells, used for the evaluation of gastrointestinal permeability in vitro. The aim of this work is to evaluate the cytotoxic and cell viability effects in a intestinal epithelium model and the antioxidant activity of FA embedded in CS matrix. The development of the mixed matrices consisted on the addition of CS to FA solution at pH 7,4.The antioxidant capacity of FA-CS mixed matrixes were evaluated by FRAP assay conducted according to the method of (Martínez et al., 2019) and ABTS assay according to the method of (Re et al., 1999). The FA antioxidant capacity was highly increased when was complexed with CS by both methodologies. Argentinian CS showed antioxidant capacity by itself only when evaluated by FRAP assay. Survival and cell proliferation were evaluated by the MTT assay, which allows to determine the mitochondrial functionality of the treated cells. Caco-2 cells were seeded at a density 2.104/well in 96 well cell culture plates and pre-incubated for 24 hours (Kowapradit et al., 2010). The cells were then treated with high and low molecular (control) weight CS (50-1000 ug/ml), FA (0.05-1.5 ug/ml) and both types of matrices containing CS (250 ug/ml) with variable ratio of FA (0.05-1.5 ug / ml) in 2% FBS medium for 24 h. The results indicated that Caco-2 cells significantly increase their proliferation respect to the control when were incubated with the FA-CS treatments.These formulated matrices could be functional ingredients for FA, protecting the vitamin against light and oxygen, and favouring its bioavailability when ingested.