Anti-inflammatory effect of two pomegranate seed oils obtained by green technologies in Caco-2 cells using the bioaccessible fraction from in vitro gastrointestinal digestion

Abstract

Pomegranate seeds contain up to 20% oil with a high content of punicic acid (85%), which is responsible for several biological activities. In this work, two pomegranate oils obtained by a two-step sequential extraction, first with an expeller and then via supercritical CO₂ technologies, have been studied in a static gastrointestinal in vitro digestion model to evaluate their bioaccessibility. The micellar phases obtained were evaluated by an in vitro model of intestinal inflammation and Caco-2 cells exposed to the inflammatory mediator lipopolysaccharide (LPS). Inflammatory response was assessed by measuring the production of interleukins IL-6 and IL-8, and tumor necrosis factor α (TNF-α), and by evaluating the monolayer integrity. The results obtained indicate that expeller pomegranate oil (EPO) provides the highest amount of micellar phase (ca. 93%) with free fatty acids and monoacylglycerols as major components. The micellar phase obtained with supercritical CO₂ pomegranate oil (SCPO) is ca. 82% with similar lipid composition. Micellar phases of EPO and SCPO showed high stability and adequate particle size. EPO shows an anti-inflammatory response, reducing the production of IL-6, IL-8 and TNF-α in LPS stimulated caco-2 cells and increasing the integrity of the cell monolayer as measured by transepithelial electrical resistance (TEER). In the case of SCPO, the anti-inflammatory effect was only evident for IL-8. The present work demonstrates good digestibility, bioaccessibility and anti-inflammatory response of both EPO and SCPO oils.

Keywords: Anti-inflammatory activity; Bioaccessible fraction; Caco-2; Conjugated linolenic acids; Gastrointestinal in vitro digestion; Green technologies; Micellar phase; Pomegranate oil.