02627nas a2200181   4500008003900000245010200039210006900141260003600210300000800246520196600254653001902220100001402239700001402253700001902267700001802286700001802304856012302322       2014                          d00aMulti-organ-on-plate system for in-vitro studies of intestinal drug absorption and hepatotoxicity0 aMultiorganonplate system for invitro studies of intestinal drug   aEgmond aan Zee, The Netherlands  a1073 a<p>More meaningful in-vitro models which simulate the physiological conditions of native tissue are becoming essential in the pharmaceutical field, for early and rapid screening of drug candidates. Here, we describe a multi-organ-on-plate system based on single and double flow mini bioreactor modules for dynamic in-vitro studies of intestinal drug absorption, drug metabolism and more relevant toxicity studies. The double flow module for membrane culture was firstly characterized using computational fluid dynamic models and measurements of pressure gradients, in order to indentify the optimal flow rates for maximizing the passage of solutes through the membrane. Then, cell culture experiments were performed with fully differentiated Caco-2 cells seeded on the semi-permeable membrane as a dynamic model of the intestinal epithelium, connected to a single flow chamber with metabolically competent human upcyteÂ® hepatocytes (Medicyte GmbH, Germany) seeded on a 3D collagen cryogel. First we assessed the role of flow in modulating the passage of compounds across the epithelial barrier. Then toxicity tests were performed by administering different concentrations of hepatotoxic compounds (i.e. Diclofenac, Nimesulide, industrial nanoparticles) in the apical compartment of the MB, compared the data with cell cultures in transwells. Our results show: i) the presence of flow significantly increases translocation of all molecules tested across the membrane, ii) flow conditioned Caco-2 cells are more permeable to small hydrophilic compounds, despite having high TEER values iii) although they display higher levels of phenotypic markers (tight junctions, albumin expression etc), cells in the system are more susceptible to drug induced toxicity. In conclusion, the multi-organ-on-plate system predicts drug adsorption and toxicity better than traditional cell cultures and could be used to reduce, refine and eventually replace animal tests.</p>
10aBioengineering1 aGiusti, S1 aSbrana, T1 aGiacopelli, D.1 aDi Patria, V.1 aAhluwalia, A.  uhttp://95.110.227.16/publications/multi-organ-plate-system-vitro-studies-intestinal-drug-absorption-and-hepatotoxicity02045nas a2200205   4500008004100000022001400041245010800055210006900163520140900232653001901641100001401660700001401674700001601688700001801704700001901722700001501741700001701756700001801773856004801791       2014                        eng d  a1860-731400aA novel dual-flow bioreactor simulates increased fluorescein permeability in epithelial tissue barriers0 anovel dualflow bioreactor simulates increased fluorescein permea3 a<p>Permeability studies across epithelial barriers are of primary importance in drug delivery as well as in toxicology. However, traditional in vitro models do not adequately mimic the dynamic environment of physiological barriers. Here, we describe a novel two-chamber modular bioreactor for dynamic in vitro studies of epithelial cells. The fluid dynamic environment of the bioreactor was characterized using computational fluid dynamic models and measurements of pressure gradients for different combinations of flow rates in the apical and basal chambers. Cell culture experiments were then performed with fully differentiated Caco-2 cells as a model of the intestinal epithelium, comparing the effect of media flow applied in the bioreactor with traditional static transwells. The flow increases barrier integrity and tight junction expression of Caco-2 cells with respect to the static controls. Fluorescein permeability increased threefold in the dynamic system, indicating that the stimulus induced by flow increases transport across the barrier, closely mimicking the in vivo situation. The results are of interest for studying the influence of mechanical stimuli on cells, and underline the importance of developing more physiologically relevant in vitro tissue models. The bioreactor can be used to study drug delivery, chemical, or nanomaterial toxicity and to engineer barrier tissues.</p>
10aBioengineering1 aGiusti, S1 aSbrana, T1 aLa Marca, M1 aDi Patria, V.1 aMartinucci, V.1 aTirella, A1 aDomenici, C.1 aAhluwalia, A.  uhttp://www.ncbi.nlm.nih.gov/pubmed/24756869