From mouth to gut: microfluidic in vitro simulation of human gastro-intestinal digestion and intestinal permeability.

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 (Papp) of Lucifer Yellow closely approximating values reported ex vivo (3.7 × 10-6 ± 1.4 × 10-6vs. 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.

Potassium urinary excretion and dietary intake: a cross-sectional analysis in 8-10 year-old children.

BACKGROUND: Data from studies assessing the intake of potassium, and the concomitant sodium-to-potassium ratio are limited. The aim of this study was to evaluate potassium and sodium-to-potassium ratio intake in 8-10 year-old children. METHODS: A cross-sectional survey was carried out from January to June 2014 and data from 163 children (81 boys) were included. Potassium intake was estimated by 24-h urine collection and coefficient of creatinine was used to validate completeness of urine collections. Urinary sodium and sodium-to-potassium ratio were also analysed. A 24-h dietary recall was used to provide information on dietary sources of potassium. Height and weight were measured according to international standards. RESULTS: The mean urinary potassium excretion was 1701 ± 594 mg/day in boys, and 1682 ± 541 mg/day in girls (p = 0.835); 8.0% of children met the WHO recommendations for potassium intake. The mean sodium excretion was 2935 ± 1075 mg/day in boys and 2381 ± 1045 mg/day in girls (p <0.001) and urinary sodium-to-potassium ratio was 3.2 ± 1.4 in boys, and 2.5 ± 1.1 in girls (p = 0.002). The mean fruit and vegetable intake was 353.1 ± 232.5 g/day in boys, and 290.8 ± 213.1 g/day in girls (p = 0.101). CONCLUSIONS: This study reported a low compliance of potassium intake recommendations in 8-10 year-old children. Health promotion interventions are needed in order to broaden public awareness of potassium inadequacy and to increase potassium intake.

Resveratrol-loaded octenyl succinic anhydride modified starch emulsions and hydroxypropyl methylcellulose (HPMC) microparticles: Cytotoxicity and antioxidant bioactivity assessment after in vitro digestion.

Hydroxypropyl methylcellulose (HPMC)-based microparticles and modified starch emulsions (OSA-MS) were loaded with resveratrol and characterized regarding their physicochemical and thermal properties. Both delivery systems were subject to an in vitro gastrointestinal digestion to assess the bioaccessibility of resveratrol. In addition, cell-based studies were conducted after in vitro digestion and cytotoxicity and oxidative stress were assessed. HPMC-based microparticles displayed higher average sizes (d) and lower polydispersity index (PDI) (d = 948 nm, PDI < 0.2) when compared to OSA-MS-based emulsions (d = 217 nm, PDI < 0.3). Both proved to protect resveratrol under digestive conditions, leading to an increase in bioaccessibility. Resveratrol-loaded HPMC-microparticles showed a higher bioaccessibility (56.7 %) than resveratrol-loaded emulsions (19.7 %). Digested samples were tested in differentiated co-cultures of Caco-2 and HT29-MTX, aiming at assessing cytotoxicity and oxidative stress, and a lack of cytotoxicity was observed for all samples. Results displayed an increasing antioxidant activity, with 1.6-fold and 1.4-fold increases over the antioxidant activity of free resveratrol, for HPMC-microparticles and OSA-MS nanoemulsions, respectively. Our results offer insight into physiological relevancy due to assessment post-digestion and highlight the protection that the use of micro-nano delivery systems can confer to resveratrol and their potential to be used as functional food ingredients capable of providing antioxidant benefits upon consumption.

Recent Advances in Cell-Based In Vitro Models to Recreate Human Intestinal Inflammation.

Inflammatory bowel disease causes a major burden to patients and healthcare systems, raising the need to develop effective therapies. Technological advances in cell culture, allied with ethical issues, have propelled in vitro models as essential tools to study disease aetiology, its progression, and possible therapies. Several cell-based in vitro models of intestinal inflammation have been used, varying in their complexity and methodology to induce inflammation. Immortalized cell lines are extensively used due to their long-term survival, in contrast to primary cultures that are short-lived but patient-specific. Recently, organoids and organ-chips have demonstrated great potential by being physiologically more relevant. This review aims to shed light on the intricate nature of intestinal inflammation and cover recent works that report cell-based in vitro models of human intestinal inflammation, encompassing diverse approaches and outcomes.

Whey-pectin microcapsules improve the stability of grape marc phenolics during digestion.

Grape marc (GM) is an agri-food residue from the wine industry valuable for its high content of phenolic compounds. This study aimed to develop an encapsulation system for GM extract (GME) using food-grade biopolymers resistant to gastric conditions for its potential use as a nutraceutical. For this purpose, a hydroalcoholic GME was prepared with a total phenolics content of 219.62 ± 11.50 mg gallic acid equivalents (GAE)/g dry extract and 1389.71 ± 97.33 µmol Trolox equivalents/g dry extract antioxidant capacity, assessed through ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) assay. Moreover, the extract effectively neutralized reactive oxygen species in Caco-2 cells, demonstrating an intracellular antioxidant capacity comparable to Trolox. The GME was encapsulated using whey protein isolate and pectin through nano spray drying (73% yield), resulting in spherical microparticles with an average size of 1 ± 0.5 µm and a polydispersity of 0.717. The encapsulation system protected the microcapsules from simulated gastrointestinal digestion (GID), where at the end of the intestinal phase, 82% of the initial phenolics were bioaccessible compared to 54% in the free GME. Besides, the encapsulated GME displayed a higher antioxidant activity by the ferric reducing antioxidant power assay than the free extract after GID. These results show the potential of this encapsulation system for applying GME as a nutraceutical with a high antioxidant capacity and protective effect against cellular oxidation.

Anti-Inflammatory Protein Isolated from Tamarind Promotes Better Histological Aspects in the Intestine Regardless of the Improvement of Intestinal Permeability in a Preclinical Study of Diet-Induced Obesity.

Obesity is associated with metabolic and physiological effects in the gut. In this study, we evaluated the anti-inflammatory effect of trypsin inhibitor isolated from tamarind seeds (TTI) in vitro (interaction with lipopolysaccharide (LPS) and inhibitory activity against human neutrophil elastase (HNE)), and using intestinal co-cultures of Caco-2:HT29-MTX cell lines inflamed with TNF-α (50 ng/mL) and a Wistar rat model of diet-induced obesity (n = 15). TTI was administered to animals by gavage (10 days), and the treated group (25 mg/kg/day) was compared to animals without treatment or treated with a nutritionally adequate diet. In the in vitro study, it showed inhibitory activity against HNE (93%). In co-cultures, there was no protection or recovery of the integrity of inflamed cell monolayers treated with TTI (1.0 mg/mL). In animals, TTI led to lower plasma concentrations of TNF-α and IL-6, total leukocytes, fasting glucose, and LDL-c (p < 0.05). The intestines demonstrated a lower degree of chronic enteritis, greater preservation of the submucosa, and greater intestinal wall thickness than the other groups (p = 0.042). Therefore, the better appearance of the intestine not reflected in the intestinal permeability added to the in vitro activity against HNE point to possibilities for new studies and applications related to this activity.

Fabrication of Quasi-2D Shape-Tailored Microparticles using Wettability Contrast-Based Platforms.

The ability to fabricate materials with ultrathin architectures enables the breakthrough of low-dimensional structures with high surface area that showcase distinctive properties from their bulk counterparts. They are exploited in a wide range of fields, including energy harvesting, catalysis, and biomedicine. Despite such versatility, the fine tuning of the lateral dimensions and geometry of these structures remains challenging. Prepatterned platforms gain significant attention as enabling technologies to process materials with highly controlled shapes and dimensions. Herein, different nanometer-thick particles of various lateral sizes and geometries (e.g., squares, circles, triangles, hexagons) are processed with high precision and definition, taking advantage of the wettability contrast of oleophilic-oleophobic patterned surfaces. Quasi-2D polymeric microparticles with high shape- and size-fidelity can be retrieved as freestanding objects in a single step. These structures show cell-mediated pliability, and their integration in gravity-enforced human adipose-derived stem cell spheroids leads to an enhanced metabolic activity and a modulated secretion of proangiogenic factors.

Microparticles in Contact with Cells: From Carriers to Multifunctional Tissue Modulators.

For several decades microparticles have been exclusively and extensively explored as spherical drug delivery vehicles and large-scale cell expansion carriers. More recently, microparticulate structures gained interest in broader bioengineering fields, integrating myriad strategies that include bottom-up tissue engineering, 3D bioprinting, and the development of tissue/disease models. The concept of bulk spherical micrometric particles as adequate supports for cell cultivation has been challenged, and systems with finely tuned geometric designs and (bio)chemical/physical features are current key players in impacting technologies. Herein, we critically review the state of the art and future trends of biomaterial microparticles in contact with cells and tissues, excluding internalization studies, and with emphasis on innovative particle design and applications.

Urinary hydration biomarkers and dietary intake in children.

INTRODUCTION: The importance of hydration is undoubtable but reliable data on hydration status and its relation with diet is lacking. OBJECTIVES: We aimed to evaluate the hydration status and its relation to beverages and food intake in children. METHODS: A sample of 172 (50% male), 7-11 year-old children was included in this survey. Participants completed a 24 h urine collection; a 24 hours food recall corresponding to the day of urine collection was applied, weight and height were measured and parents/caregivers fi lled a lifestyle and socio-demographic questionnaire. The free water reserve was used to assess the hydration status. The intakes of food and beverages were compared according to hydration status using the t-test, Mann-Whitney test or unconditional regression models as appropriate. RESULTS: More than half of the participants were classifi ed as hypohydrated or at risk of hypohydration (57% in girls and 58% in boys). Compared to hypohydrated children, a signifi cant higher consumption of water (276.2 ± 208.4 vs 188.2 ± 187.4 g/day) and fruit juices (77.6 ± 139.4 vs 14.4 ± 57.2 g/day) was reported by euhydrated boys and girls, respectively. Lower consumers of water and fruit juices showed a higher risk of hypohydration (OR = 2.16, 95% CI: 1.02-4.58, p = 0.045), adjusting for confounders. CONCLUSIONS: Most of the children included in this analysis were classifi ed as at risk of hypohydration and those with higher water and fruit juices consumption showed a better hydration status.

Urinary hydration biomarkers and dietary intake in children

Introduction: The importance of hydration is undoubtable but reliable data on hydration status and its relation with diet is lacking. Objectives: We aimed to evaluate the hydration status and its relation to beverages and food intake in children. Methods: A sample of 172 (50% male), 7-11 year-old children was included in this survey. Participants completed a 24 h urine collection; a 24 hours food recall corresponding to the day of urine collection was applied, weight and height were measured and parents/caregivers filled a lifestyle and socio-demographic questionnaire. The free water reserve was used to assess the hydration status. The intakes of food and beverages were compared according to hydration status using the t-test, Mann-Whitney test or unconditional regression models as appropriate. Results: More than half of the participants were classified as hypohydrated or at risk of hypohydration (57% in girls and 58% in boys). Compared to hypohydrated children, a significant higher consumption of water (276.2 ± 208.4 vs 188.2 ± 187.4 g/day) and fruit juices (77.6 ± 139.4 vs 14.4 ± 57.2 g/day) was reported by euhydrated boys and girls, respectively. Lower consumers of water and fruit juices showed a higher risk of hypohydration (OR = 2.16, 95% CI: 1.02-4.58, p = 0.045), adjusting for confounders. Conclusions: Most of the children included in this analysis were classified as at risk of hypohydration and those with higher water and fruit juices consumption showed a better hydration status (AU)

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