This study investigates the gut anti-inflammatory activity of a plant sterol (PS) food supplement (PS-FS), alongside PS-enriched milk-based fruit beverage and PS-enriched rye bread. A co-culture model based on a dual-chamber system with differentiated intestinal-like Caco-2 cells (apical) and RAW264.7 macrophages (basolateral) was used. The bioaccessible fractions (BF) of the samples were obtained after INFOGEST 2.0 simulated gastrointestinal digestion. The BF were added to the apical part (diluted 1/20 v/v with culture medium to avoid cytotoxicity) for 90 min, followed by stimulation with lipopolysaccharide (LPS) (1 µg mL^{– 1}, 24 h) on the basolateral side. The pharmacological interaction between samples and budesonide (1 µM/90 min) was evaluated. Results indicate that PS-FS significantly attenuated LPS-induced secretion of IL-8 (28%) by Caco-2 cells, and TNF-α (9%) and IL-6 (54%) by RAW264.7 macrophages, whereas PS-enriched beverage and bread did not exhibit protective effects. Additionally, PS-FS demonstrated an improvement in oxidative status in Caco-2 cells, evidenced by reduced levels of reactive oxygen species (47%), iNOS protein expression (27%), and nitrite/nitrate secretion (27%). Mechanistically, PS-FS inhibited the NF-B-COX-2-PGE2 signaling pathway in macrophages, resulting in decreased NF-κB p65 nuclear translocation (39%), COX-2 protein expression (32%), and PGE2 production (27%). Co-treatment with budesonide and PS-FS displayed an antagonistic effect (combination index 0.38-0.63). This study demonstrates the potent intestinal anti-inflammatory activity of a PS-FS, positioning it as a promising nutraceutical product for the management of inflammatory bowel diseases. However, the food matrix of the milk-based fruit beverage and rye bread appear to interfere with the anti-inflammatory activity of PS.

Bioaccessibility of plant sterols (PS) in an enriched wholemeal rye bread was evaluated, for the first time, using the INFOGEST protocol without gastric lipase (GL) and cholesterol esterase (CE), with GL or GL + CE. Moreover, human chewing and an in vitro oral phase (simulated salivary fluid and α-amylase) were evaluated for this purpose. The addition of GL decreased the bioaccessibility of total PS (from 23.8 to 18.5%), whereas the use of GL + CE does not significantly affect PS bioaccessibility. The in vitro oral phase resulted in an ineffective homogenization of the fresh vs partially dried and milled bread, reducing the bioaccessibility of total (from 20.2 to 12.8%) and individual PS. The INFOGEST digestion including the use of GL and CE, as well as an oral phase with human chewing, is proposed for the assessment of PS bioaccessibility in a solid matrix such as wholemeal rye bread since it more closely approximates the in vivo situation.

Bread is one of the staple foods of many countries, and its enrichment with bioactive compounds is trending. This phenomenon is focused on breads with a good nutritional profile, such as wholemeal rye bread (WRB), in which enrichment with plant sterols (PSs) is allowed in accordance with European regulations. The objective of the present study was to optimize the production of a WRB enriched with PS (PS-WRB) and to evaluate the proximate composition and starch digestibility as an indicator of nutritional quality. The rheological analysis showed that the bread dough presents satisfactory farinographic properties (dough development time 6 min; stability 4 min; degree of softening 100 Brabender units) but high water absorption (67%). The PS-WRB is high in dietary fiber and low in protein (20.4 and 7.7% w/w, dry basis, respectively) compared with other cereals reported in the scientific literature. In turn, a low starch proportion was hydrolyzed during the simulated digestion (59.9% of total starch), being also slowly hydrolyzed, as deduced from the rapidly digestible starch value (56.5% of total starch). In conclusion, WRB is a suitable matrix for PS enrichment, which allows for obtaining a product with a good nutritional profile and potential health benefits.

This in vitro study reveals that ethylcoprostanol, a β-sitosterol metabolite produced by gut microbiota, reduces colon cancer cell proliferation and protects non-tumor cells from cholesterol metabolite-induced toxicity.

The prevention of cardiovascular disease using foods fortified with plant sterols (PS), with a hypocholesterolemic effect, is important for the elderly population.