Stéphanie Blanquet-Diot

Relevance and challenges in modeling human gastric and small intestinal digestion

Gastric and small intestinal (GSI) models are increasingly used as an alternative to in vivo assays to answer many questions raised by industry and researchers. A broad range of in vitro systems is available, from static monocompartmental to dynamic multicompartmental models. However, these models require a compromise between technological complexity and biological significance. Further efforts and technological innovations are still needed to improve in vitro models and meet growing demands in the areas of nutrition and health. This review describes the models available to date for the human stomach and small intestine and highlights their relevance in nutritional, toxicological, pharmaceutical, and microbiological studies. Limitations and challenges facing artificial digestion technology are also discussed.

Digestive Stability of Xanthophylls Exceeds That of Carotenes As Studied in a Dynamic in Vitro Gastrointestinal System

Epidemiological studies have suggested that high consumption of tomato products is associated with a lower risk for chronic diseases. To exert their health effect, the phytochemicals of tomatoes have to be bioavailable and therefore it implies their stability through the digestion process. Here, we assessed the digestive stability of the red-pigmented lycopene and other carotenoids brought in nutritional quantity within different food matrixes, using the TNO gastrointestinal tract model (TIM). This multicompartmental dynamic system accurately reproduces the main parameters of gastric and small intestinal digestion in human. In vitro digestions of a standard meal containing red tomato (RT), yellow tomato (devoid of lycopene), or lycopene beadlets were performed. Zeaxanthin and lutein were stable throughout artificial digestions, whereas beta-carotene and all-trans lycopene were degraded (approximately 30 and 20% loss at the end of digestion, respectively) in the jejunal and ileal compartments. The recovery of beta-carotene in the digesta of the RT meal was significantly lower than that in the yellow one, showing a food matrix effect. In the same way, until 180 min of digestion, the recovery percentages of all-trans lycopene from RT were significantly lower than those issued from the supplement. Isomeric conformation also influenced the stability of carotenoids, 5-cis lycopene being the most stable isomer followed by all-trans and 9-cis. No trans-cis isomerization of lycopene occurred in the TIM. By using a relevant dynamic in vitro system, this study allowed us to gain further insight into the parameters influencing the digestive stability of carotenoids, and therefore their bioavailability, in humans.

Combining the dynamic TNO-gastrointestinal tract system with a caco-2 cell culture model: application to the assessment of lycopene and α-tocopherol bioavailability from a whole food.

To exert their health effect, phytochemicals such as carotenoids and vitamin E have to be bioavailable. We investigated the digestive stability and intestinal absorption of lycopene and alpha-tocopherol from a whole food containing red tomatoes and sunflower oil using, for the first time, the dynamic gastrointestinal system TNO gastrointestinal tract model (TIM) coupled with Caco-2 cells. Digestive samples were added to Caco-2 cells after appropriate ultracentrifugation, filtration, and dilution. alpha-Tocopherol was stable during digestion in the TIM, whereas a 25% loss was observed for lycopene. The absorption of both compounds was curvilinear, bidirectional, and concentration-dependent. The percentages of alpha-tocopherol absorbed, but not that of lycopene, were lower with digestas compared to those with pure compounds, suggesting competition for absorption with other components of the test meal. According to in vivo data, a lower bioavailability was found for lycopene compared to that for alpha-tocopherol. These results support the usefulness of this in vitro approach for estimating the bioavailability of active compounds from food.

Effect of a New Probiotic Saccharomyces cerevisiae Strain on Survival of Escherichia coli O157:H7 in a Dynamic Gastrointestinal Model

ABSTRACT Survival of Escherichia coli O157:H7 was investigated using a dynamic gastrointestinal model. A high bacterial mortality was observed in the stomach and duodenum. In contrast, bacteria grew in the distal parts of the small intestine. The coadministration of Saccharomyces cerevisiae CNCM I-3856 led to a significant reduction of bacterial resumption, maybe through ethanol production.

Interactions with M Cells and Macrophages as Key Steps in the Pathogenesis of Enterohemorragic Escherichia coli Infections

Enterohemorrhagic Escherichia coli (EHEC) are food-borne pathogens that can cause serious infections ranging from diarrhea to hemorrhagic colitis (HC) and hemolytic-uremic syndrome (HUS). Translocation of Shiga-toxins (Stx) from the gut lumen to underlying tissues is a decisive step in the development of the infection, but the mechanisms involved remain unclear. Many bacterial pathogens target the follicle-associated epithelium, which overlies Peyers patches (PPs), cross the intestinal barrier through M cells and are captured by mucosal macrophages. Here, translocation across M cells, as well as survival and proliferation of EHEC strains within THP-1 macrophages were investigated using EHEC O157:H7 reference strains, isogenic mutants, and 15 EHEC strains isolated from HC/HUS patients. We showed for the first time that E. coli O157:H7 strains are able to interact in vivo with murine PPs, to translocate ex vivo through murine ileal mucosa with PPs and across an in vitro human M cell model. EHEC strains are also able to survive and to produce Stx in macrophages, which induce cell apoptosis and Stx release. In conclusion, our results suggest that the uptake of EHEC by M cells and underlying macrophages in the PP may be a critical step in Stx translocation and release in vivo. A new model for EHEC infection in humans is proposed that could help in a fuller understanding of EHEC-associated diseases.

Comparison of a static and a dynamic in vitro model to estimate the bioaccessibility of As, Cd, Pb and Hg from food reference materials Fucus sp. (IAEA-140/TM) and Lobster hepatopancreas (TORT-2)

Bioaccessibility, the fraction of an element solubilized during gastrointestinal digestion and available for absorption, is a factor that should be considered when evaluating the health risk of contaminants from food. Static and dynamic models that mimic human physiological conditions have been used to evaluate bioaccessibility. This preliminary study compares the bioaccessibility of arsenic (As), cadmium (Cd), lead (Pb) and mercury (Hg) in two food certified reference materials (CRMs) (seaweed: Fucus sp., IAEA-140/TM; Lobster hepatopancreas: TORT-2), using two in vitro gastrointestinal digestion methods: a static method (SM) and a dynamic multicompartment method (TIM-1). There are significant differences (p<0.05) between the bioaccessible values of As, Cd, Pb and Hg obtained by SM and TIM-1 in the two CRMs. The specific form in which the elements studied are present in the CRM may help to explain the bioaccessibility values obtained.

Use of Artificial Digestive Systems to Investigate the Biopharmaceutical Factors Influencing the Survival of Probiotic Yeast During Gastrointestinal Transit in Humans

PurposeTo evaluate the influence of the main biopharmaceutical factors on the viability of a new probiotic yeast strain, using dynamic in vitro systems simulating human gastric/small intestinal (TIM) and large intestinal (ARCOL) environments.MethodsThe viability of Saccharomyces cerevisiae CNCM I-3856 throughout the artificial digestive tract was determined by microbial counting. We investigated the effects of galenic formulation, food intake, dose, mode and frequency of administration on yeast survival rate.ResultsIn both fasted and fed states, yeast viability in the upper digestive tract was significantly higher when the probiotic was administered in hydroxypropylmethylcellulose (HPMC) capsules compared to tablets. Food intake led to a delay in yeast release and a two-fold increase in strain survival. Whatever the dose, yeasts were particularly sensitive to the large intestinal environment. High concentrations of probiotic could only be maintained in the colon when it was inoculated twice a day over a 5-h-period.ConclusionsTIM and ARCOL are complementary in vitro tools relevant for screening purposes, supplying valuable information on the effects of galenic form, food intake and dose regimen on the viability of probiotics throughout the human digestive tract.

Gut Microbiota Dysbiosis in Postweaning Piglets: Understanding the Keys to Health

Weaning is a critical event in the pigs life cycle, frequently associated with severe enteric infections and overuse of antibiotics; this raises serious economic and public health concerns. In this review, we explain why gut microbiota dysbiosis, induced by abrupt changes in the diet and environment of piglets, emerges as a leading cause of post-weaning diarrhea, even if the exact underlying mechanisms remain unclear. Then, we focus on nonantimicrobial alternatives, such as zinc oxide, essential oils, and prebiotics or probiotics, which are currently evaluated to restore intestinal balance and allow a better management of the crucial weaning transition. Finally, we discuss how in vitro models of the piglet gut could be advantageously used as a complement to ex vivo and in vivo studies for the development and testing of new feed additives.

Enterohemorrhagic Escherichia coli O157:H7 Survival in an In Vitro Model of the Human Large Intestine and Interactions with Probiotic Yeasts and Resident Microbiota

ABSTRACT This is the first report on the fate of enterohemorrhagic Escherichia coli O157:H7 in simulated human colonic conditions. The pathogen was progressively eliminated from the bioreactor and did not modify the major populations of resident microbiota. The coadministration of the Saccharomyces cerevisiae CNCM I-3856 probiotic strain led to a significant increase in acetate production but did not reduce pathogen viability.

Survival of Escherichia coli O26:H11 exceeds that of Escherichia coli O157:H7 as assessed by simulated human digestion of contaminated raw milk cheeses.

Shiga toxin producing Escherichia coli (STEC) are an important cause of human foodborne outbreaks. The consumption of raw milk dairy products may be an important route of STEC infection. For successful foodborne transmission, STEC strains must survive stress conditions met during gastrointestinal transit in humans. The aim of this study was to evaluate the survival of two STEC strains of serotypes O157:H7 and O26:H11 during simulated human digestion in the TNO gastro-Intestinal tract Model (TIM) of contaminated uncooked pressed cheeses. The survival of cheese microflora during in vitro gastrointestinal transit was also determined for the first time. The level of STEC increased from 2 log₁₀ CFU/ml to 4 log₁₀ CFU/g during the first 24h of cheese making and remained stable at around 4 log₁₀ CFU/g during cheese ripening and conservation. During transit through the artificial stomach and duodenum, levels of STEC decreased: 0.2% of E. coli O157:H7 and 1.8% of E. coli O26:H11 were recovered at 150 min in the gastric compartment, compared with 14.3% for the transit marker. Bacterial resumption was observed in the jejunum and ileum: 35.8% of E. coli O157:H7 and 663.2% of E. coli O26:H11 were recovered at 360 min in the ileal compartment, compared with 12.6% for the transit marker. The fate of STEC was strain-dependent, the survival of E. coli O26:H11 being 13 times greater than that of E. coli O157:H7 at the end of digestion in the cumulative ileal deliveries. These data provide a better understanding of STEC behavior during gastrointestinal transit in humans after ingestion of contaminated cheese.