A multicompartmental bioreactor was conceived and designed to mimic cross talk between cells in different culture chambers connected only by flow, such that cell-cell interaction is mediated by soluble ligands as occurs in the body. The system was tested with a connected culture of murine hepatocytes and human umbilical vein endothelial cells. Metabolites such as albumin, urea, lactate and viability were monitored during the course of the experiments and compared with monoculture conditions in the bioreactor. When the two cell types are placed in connected culture, there is an increase in endothelial cell viability and hepatic glucose synthesis as well as albumin and urea production, while overall lactate production in the system is downregulated. The results show that the multicompartmental bioreactor enhances cell function, effectively combining both heterotypic interactions with increased nutrient availability.

The liver and other organs are connected to each other through the bloodstream. Therefore, the connection between tissues is generally mediated by soluble molecules able to cross the endothelial wall of capillaries. We developed a multicompartmental device, multicompartmental bioreactor (MCB), designed to mimic the connection between different tissues in which crosstalk is mediated by soluble molecules transported through the blood. A comparative study of the crosstalk between hepatocytes (HepG2) and endothelial cells (human umbilical vein endothelial cells) in connected culture in the MCB and in a traditional static coculture system was performed by analyzing glucose consumption and secretion of albumin, urea, and nitric oxide. When hepatocytes and endothelial cells were cultured together, the production of albumin and urea increased, and the increase was higher in the MCB than in traditional static coculture. In spite of this enhanced metabolic activity, the crosstalk between hepatocytes and endothelial cell leads to decreased glucose consumption with respect to hepatocytes alone, both in static and in dynamic conditions. However, the dynamic connected culture has a higher rate of metabolite synthesis and secretion with respect to cocultures. This means a more efficient use of energetic substrates and enhanced hepatocyte function in the MCB.

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Blends between chitosan (CS) and gelatin (G) with various compositions (CS/G 0/100 20/80, 40/60, 60/40, 100/0 w/w) were produced as candidate materials for biomedical applications. Dehydro-thermal crosslinking was adopted to promote the formation of amide and ester bonds between the macromolecules ((CS/G)-t). The effect of composition and crosslinking on the physico-chemical properties of the samples was evaluated by scanning electron microscopy, thermogravimetry, contact angle measurements, dissolution and swelling tests. Mechanical properties of (CS/G)-t samples were also determined through stress-strain and creep-recovery tests. The elastic moduli of dry blend samples showed a positive deviation from the additive law of the in-series model, because of interactions and/or chemical bonds between components. The comparison between the elastic moduli of wet samples and those of different human tissues showed that (CS/G)-t substrates can be suitable for soft-tissue reconstruction. (CS/G)-t two-dimensional scaffolds were fabricated by micro-molding, based on the use of a polydimethylsiloxane mould to create patterns with micro-scale resolution on cast films. Biocompatibility of (CS/G)-t samples was studied by means of cell tests using NIH-3T3 fibroblasts. Finally, the evaluation of the affinity of (CS/G)-t samples towards neuroblastoma cells adhesion and proliferation was performed, showing promising results for the blend containing 80 wt % gelatin.

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To develop in vitro models of cells, tissues and organs we have designed and realized a series of cell culture chambers. Each chamber is purpose designed to simulate a particular feature of the in vivo environment. The bioreactor system is user friendly, and the chambers are easy to produce, sterilize and assemble. In addition they can be connected together to simulate inter-organ or tissue cross-talk. Here we discuss the design philosophy of the bioreactor system and then describe its construction. Preliminary results of validation tests obtained with hepatocytes and endothelial cells are also reported. The results show that endothelial cells are extremely sensitive to small levels of shear stress and that the presence of heterotypic signals from endothelial cells enhances the endogenous metabolic function of hepatocytes.

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Abundant experimental evidence demonstrates that endothelial cells are sensitive to flow; however, the effect of fluid pressure or pressure gradients that are used to drive viscous flow is not well understood. There are two principal physical forces exerted on the blood vessel wall by the passage of intra-luminal blood: pressure and shear. To analyze the effects of pressure and shear independently, these two stresses were applied to cultured cells in two different types of bioreactors: a pressure-controlled bioreactor and a laminar flow bioreactor, in which controlled levels of pressure or shear stress, respectively, can be generated. Using these bioreactor systems, endothelin-1 (ET-1) and nitric oxide (NO) release from human umbilical vein endothelial cells were measured under various shear stress and pressure conditions. Compared to the controls, a decrease of ET-1 production by the cells cultured in both bioreactors was observed, whereas NO synthesis was up-regulated in cells under shear stress, but was not modulated by hydrostatic pressure. These results show that the two hemodynamic forces acting on blood vessels affect endothelial cell function in different ways, and that both should be considered when planning in vitro experiments in the presence of flow. Understanding the individual and synergic effects of the two forces could provide important insights into physiological and pathological processes involved in vascular remodeling and adaptation.