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Reproducible in vitro studies of bioaccessibility, intestinal absorption, and bioavailability are key to the successful development of novel food ingredients or drugs intended for oral administration. There is currently a lack of methods that offer the finesse required to study these parameters for valuable molecules typically found in small volumes - as is the case of nanomaterials, which are often used to carry and protect bioactives. Here, we describe a modular microfluidic-based platform for total simulation of the human gastro-intestinal tract. Digestion-chips and cell-based gut-chips were fabricated from PDMS by soft lithography. On-chip digestion was validated using a fluorescently labelled casein derivative, which followed typical Michaelis-Menten kinetics and showed temporal resolution and good agreement with well-established bench-top protocols. Irreversible inhibition of serine proteases using Pefabloc® SC and a 1 : 6 dilution was sufficient to mitigate the cytotoxicity of simulated digestion fluids. Caco-2/HT29-MTX co-cultures were grown on-chip under a continuous flow for 7 days to obtain a differentiated cell monolayer forming a 3D villi-like epithelium with clear tight junction formation, and with an apparent permeability ( P app ) of Lucifer Yellow closely approximating values reported ex vivo (3.7 × 10 -6 ± 1.4 × 10 -6 vs. 4.0 × 10 -6 ± 2.2 × 10 -6 ). Digesta from the digestion-chips were flowed through the gut-chip, demonstrating the capacity to study sample digestion and intestinal permeability in a single microfluidic platform holding great promise for use in pharmacokinetic studies.

Encapsulation structures for oral administration have been widely employed by the food, personal care, and pharmaceutical industries. Emulsion-filled microgels can be used to encapsulate bioactive compounds, allowing the entrapment of lipid droplets in biopolymer networks and promoting bioactive protection. The influence of pH and biopolymer concentration on the formation and structure of emulsions was evaluated, allowing the production of emulsion-filled hydrogels with potato starch as the main compound, a low alginate concentration, and gelatin in the continuous phase. Potato starch was used because it is generally recognized as safe (GRAS) and has phosphate groups, which allow electrostatic interactions with biopolymers and provide resistance to the network. Emulsion stability was achieved at pH 6, while complexation was verified under acidic conditions, which made the ionic gelation process unfeasible for the production of microgels. After defining the pH for emulsion production, microgels were formed by ionic gelation and coated microgels by electrostatic interactions, as evidenced by quartz crystal microbalance. The alginate and gelatin coating did not affect the morphology of the microparticles. An in vitro digestion assay showed that microgels composed mainly of potato starch were not degraded in the simulated mouth step. The coating layer provided extra microgel protection during digestion, demonstrating the ability of encapsulation systems to promote targeted delivery of bioactive compounds.

Grape pomace (GP) is a major byproduct worldwide, and it is well known for its bioactive compounds, such as fibers and phenolic compounds, that are popular for their impact upon human health, including gastrointestinal health. The objective of this work was to evaluate the chemical composition and biological activities of an enzymatic GP extract, as well as to investigate how gastrointestinal digestion (GID) modulates these properties. GP extract was previously produced using an enzymatic cocktail with xylanase activity and was then exposed to simulated conditions of GID, characterized for its chemical composition, and screened for antimicrobial, prebiotic, and antioxidant activities. The safety of this ingredient after GID was also assessed. GP extract presented high contents of dietary fiber and other carbohydrates, including xylooligosaccharides, in addition to minerals and phenolic compounds. In vitro simulated GID revealed that xylobiose was resistant to gastric conditions, unlike phenolic compounds. The use of 2% (w/v) of this ingredient proved to be a potential carbon source that could be fermented by Lactobacillus and Bifidobacterium spp, even after digestion. The extract also exhibited strong antioxidant and antimicrobial activities against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa; however, after GID, the antioxidant capacity decreased, and the antimicrobial capacity was strongly reduced or lost. Furthermore, the extract safety was also guaranteed on Caco-2 intestinal cells. This novel and green GP extract proved to be composed of relevant bioactive molecules, including xylooligosaccharides, polyphenols, organic acids, and minerals, which provided different biological properties; it has potential applications in the food industry such that it can be used as an ingredient in the development of new functional foods.