Non-viral gene delivery transfection profiles influence neuronal architecture in an in vitro co-culture model.

Gene delivery from tissue engineering scaffolds can induce expression of tissue inductive factors to stimulate the cellular processes required for regeneration. Transfected cells secrete diffusible proteins that can create local concentration gradients, depending on the number, distribution, and expression level of transfected cells. These gradients are linked to cellular organization and tissue architecture during embryogenesis.

Spatially patterned gene expression for guided neurite extension.

Axon pathfinding by localized expression of guidance molecules is critical for the proper development of the nervous system. In this report, we present a well-defined spatially patterned gene expression system to investigate neurite guidance in vitro. Nonviral gene delivery was patterned by combining substrate-mediated gene delivery with soft lithography techniques, and the amount of protein produced at the region of localized expression was varied by altering the vector concentration and the width of the pattern, highlighting the flexibility of the system.

Non-viral vector delivery from PEG-hyaluronic acid hydrogels.

Hydrogels have been widely used in tissue engineering as a support for tissue formation or to deliver non-viral gene therapy vectors locally. Hydrogels that combine these functionalities can provide a fundamental tool to promote specific cellular processes leading to tissue formation. This report investigates controlled release of gene therapy vectors from hydrogels as a function of the physical properties for both the hydrogel and the vector.

Patterned PLG substrates for localized DNA delivery and directed neurite extension.

Tissue engineering strategies that enable nerve regeneration will require methods that can promote and direct neurite extension across the lesion. In this report, we investigate an in vitro combinatorial approach to directed neurite outgrowth using gene delivery from topographically patterned substrates, which can induce expression of neurotrophic factors to promote neurite extension and direct the extending neurites. Poly(lactide-co-glycolide) (PLG), which has been used to fabricate conduits or bridges for regeneration, was compression molded to create channels with 100, 150, and 250 microm widths.

Spatially patterned gene delivery for localized neuron survival and neurite extension.

Natural tissues can have complex architectures, which arise in part from spatial patterns in gene expression. Regenerative strategies for damaged tissue must recreate these architectures to restore function. In this article, we demonstrate spatially controlled gene delivery from a substrate for directing cellular processes.