A Milled Microdevice to Advance Glia-Mediated Therapies in the Adult Nervous System

Abstract

Neurodegenerative disorders affect millions of adults worldwide. Neuroglia have become recent therapeutic targets due to their reparative abilities in the recycling of exogenous neurotoxins and production of endogenous growth factors for proper functioning of the adult nervous system (NS). Since neuroglia respond effectively to stimuli within in vivo environments on the micron scale, adult glial physiology has remarkable synergy with microscale systems. While clinical studies have begun to explore the reparative action of Müller glia (MG) of the visual system and Schwann Cells (ShC) of the peripheral NS after neural insult, few platforms enable the study of intrinsic neuroglia responses to changes in the local microenvironment. This project developed a low-cost, benchtop-friendly microfluidic system called the glia line system, or gLL, to advance the cellular study needed for emerging glial-based therapies. The gLL was fabricated using elastomeric kits coupled with a metal mold milled via conventional computer numerical controlled (CNC) machines. Experiments used the gLL to measure the viability, adhesion, proliferation, and migration of MG and ShC within scales similar to their respective in vivo microenvironments. Results illustrate differences in neuroglia adhesion patterns and chemotactic behavior significant to advances in regenerative medicine using implants and biomaterials, as well as cell transplantation. Data showed highest survival and proliferation of MG and ShC upon laminin and illustrated a four-fold and two-fold increase of MG migration to dosage-dependent signaling from vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF), respectively, as well as a 20-fold increase of ShC migration toward exogenous brain-derived neurotrophic factor (BDNF), compared to media control. The ability to quantify these biological parameters within the gLL offers an effective and reliable alternative to photolithography study neuroglia and their local ranges on the tens to hundreds of microns, using a low-cost and easily fabricated system.