Hybrid polymer microfluidic platform to mimic varying vascular compliance and topology.

Several cardiovascular pathologies and aging have been associated with alterations in the mechanical and structural properties of the vascular wall, leading to a reduction in arterial compliance and the development of constriction. In the past, rare efforts have been directed to understand the endothelial cell response to combined mechanical stimuli from fluid flow and substrate rigidity. Recent approaches using microfluidic platforms have limitations in precisely mimicking healthy and diseased vasculature conditions from altered topological and substrate compliance perspectives. To address this, we demonstrated an effective fabrication process to realize a hybrid polymer platform to test these mechanistic features of blood vessels. The salient features of the platform include circular microchannels of varying diameters, variation in substrate rigidity along the channel length, and the coexistence of microchannels with different cross sections on a single platform. The platform demonstrates the combined effects of flow-induced shear forces and substrate rigidity on the endothelial cell layer inside the circular microchannels. The experimental results indicate a pronounced cell response to flow induced shear stress via its interplay with the underlying substrate mechanics.

Reprogramming axonal behavior by axon-specific viral transduction.

The treatment of axonal disorders, such as diseases associated with axonal injury and degeneration, is limited by the inability to directly target therapeutic protein expression to injured axons. Current gene therapy approaches rely on infection and transcription of viral genes in the cell body. Here, we describe an approach to target gene expression selectively to axons. Using a genetically engineered mouse containing epitope-labeled ribosomes, we find that neurons in adult animals contain ribosomes in distal axons. To use axonal ribosomes to alter local protein expression, we utilized a Sindbis virus containing an RNA genome that has been modified so that it can be directly used as a template for translation. Selective application of this virus to axons leads to local translation of heterologous proteins. Furthermore, we demonstrate that selective axonal protein expression can be used to modify axonal signaling in cultured neurons, enabling axons to grow over inhibitory substrates typically encountered following axonal injury. We also show that this viral approach also can be used to achieve heterologous expression in axons of living animals, indicating that this approach can be used to alter the axonal proteome in vivo. Together, these data identify a novel strategy to manipulate protein expression in axons, and provides a novel approach for using gene therapies for disorders of axonal function.

Patterned deposition of cells and proteins onto surfaces by using three-dimensional microfluidic systems.

Three-dimensional microfluidic systems were fabricated and used to pattern proteins and mammalian cells on a planar substrate. The three-dimensional topology of the microfluidic network in the stamp makes this technique a versatile one with which to pattern multiple types of proteins and cells in complex, discontinuous structures on a surface. The channel structure, formed by the stamp when it is in contact with the surface of the substrate, limits migration and growth of cells in the channels. With the channel structure in contact with the surface, the cells stop dividing once they form a confluent layer. Removal of the stamp permits the cells to spread and divide.