Conclusions The study of the in vivo functionality of

adhering bacterial communities in the human GIT and of the localized effect on the host is frequently hindered by the complexity of reaching particular areas see more of the GIT, and by the lack of suitable in vitro models simulating the actual GIT complexity. In order to overcome this limitation we proposed the HMI module as a simplified simulation of the processes occurring at the level of the gut wall (i.e. shear stress, O2 and metabolites permeation, bacterial adhesion and host response). Three unique advantages can be ascribed to this new device, as compared to other systems available for research purposes: i) the possibility to simulate at once the bacterial adhesion to the gut wall and the indirect effect on human cell lines; ii) the possibility of performing these studies

up to 48 h with a complex microbiota, representative of that inhabiting the human gut; iii) the possibility to couple the HMI module to a continuous simulator of the human gastrointestinal tract (i.e. SHIME). The selleck kinase inhibitor latter is of key importance when analyzing the effect of specific products, as for instance prebiotic fibers. In fact, the health-modulating effect of fibers is often related to the metabolites produced by microbial species by means of cross-feeding [48, 49]. For instance, primary users often degrade part of an ingredient to smaller fragments, sugar monomers, and SCFA such as acetate or lactate. The latter two are precursors for the production of ubiquitin-Proteasome pathway the anti-inflammatory SCFA butyrate by other species [50]. The efficiency Non-specific serine/threonine protein kinase of this mechanism is frequently related to the adaptation of the microbial metabolic functionalities to the fiber and, in order to exert this effect, repeated doses of the ingredient are needed [29]. This is exactly what the combination ‘SHIME-HMI module’ allows to study: repeated doses of a product are provided to the microbiota of the SHIME; the product modifies the composition and activity of the luminal and mucosal microbiota and, ultimately, this modulates the host’s response. Several opportunities lay in the future to improve the host compartment of the

HMI module. Among them, the most challenging would be the incorporation of co-cultures of enterocytes and immune cells or of three-dimensional organotypic model of human colonic epithelium [24]. Methods The HMI module The HMI module consists of 2 compartments (each measuring 10 × 6 cm) separated by a functional double-layer composed of an upper mucus layer and a lower semi-permeable membrane (Figure 1). The upper compartment represents the luminal side of the GIT, whereas the lower compartment contains enterocytes representing the host. The polyamide membrane has a pore size of 0.2 μm and a thickness of 115 μm (Sartorius Stedim, Vilvoorde, Belgium). The mucus layer was prepared by boiling autoclaved distilled H2O containing 5% porcine mucin type II (Sigma Aldrich, St. Louis, MO, USA) and 0.8% agar. The pH was adjusted to 6.8 with 10 M NaOH.