Brain capillary endothelial cells, which constitute the blood-brain barrier (BBB), are enveloped by the extracellular matrix (ECM) produced by endothelial cells, pericytes and astrocytes. The contribution of matrix components secreted by the various cell types at the neurovascular unit, however, remains unclear with respect to their effect on endothelial barrier function. In this study, a new in vitro model was established by growing endothelial cells on an ECM produced by pericytes, astrocytes or a serial combination of both. The last-mentioned was found to be more in vivo-like. We investigated the role of the composition and morphology of ECM supra-structures in maintaining BBB function. The composition was analysed by protein analysis (enzyme-linked immunosorbent assay) and the ultrastructure of generated matrices was analysed by transmission electron microscopy including immunogold labelling. We could show by electric cell-substrate impedance sensing measurements that pericytes and combined matrices significantly improved the barrier tightness of porcine brain capillary endothelial cells (PBCEC). The increase of the resistance was verified by enhanced expression of tight junction proteins. Thus, for the first time, we have shown that barrier integrity is strictly controlled by the ECM, which is a product of all cells involved in the secretion of ECM components and their modification by corresponding cells. Moreover, we have demonstrated that complex matrices by the various cells of the BBB induce barrier marker enzymes in PBCEC, such as alkaline phosphatase.

Neural adhesion, maturation, and the correct wiring of the brain to establish each neuron's intended connectivity are controlled by complex interactions of bioactive molecules such as ligands, growth factors, or enzymes. The correct pairing of adjacent neurons is thought to be highly regulated by ligand-mediated cell-cell adhesion proteins, which are known to induce signaling activities. We developed a new platform consisting of supported lipid bilayers incorporated with Fc-chimera synaptic proteins like ephrinA5 or N-cadherin. We extensively characterized their function employing a quartz crystal microbalance with dissipation (QCM-D), calcium imaging, and immunofluorescence analysis. Our biomimetic platform has been shown to promote neural cell adhesion and to improve neural maturation at day in vitro 7 (DIV7) as indicated by an elevated expression of synaptophysin.

Microwire arrays are a powerful tool for the exertion of localized thermal stress on cellular networks. Combining microwire arrays with a set of orthogonal axon-guiding microchannels on-chip allows for the positioning of neurites, as well as control over their polarity. In this paper, we present a new fabrication approach, based on standard clean room fabrication and sacrificial layer etching, for the integration of microwire arrays into neurite guiding structures. The system permits the application of strong temperature gradients, enabling localized thermal stimulation inside microchannels.