Microfluidic systems for axonal growth and regeneration research.

Abstract

Damage to the adult mammalian central nervous system (CNS) often results in persistent neurological deficits with limited recovery of functions. The past decade has seen increasing research efforts in neural regeneration research with the ultimate goal of achieving functional recovery. Many studies have focused on prevention of further neural damage and restoration of functional connections that are compromised after injury or pathological damage. Compared to the peripheral nervous system, the failure of the adult CNS to regenerate is largely attributed to two basic aspects: inhibitory environmental influences and decreased growth capabilities of adult CNS neurons. Since early demonstration of successful growth of injured CNS axons into grafted peripheral nerve (David and Aguayo, 1981), multiple CNS axonal growth inhibitory factors have been identified and are mainly associated with degenerating CNS myelin (such as Nogo, MAG, OMgp) and with glial scar (such as chondroitin sulfate proteoglycans, CSPGs) (Yiu and He, 2006). However, blockade of these extracellular inhibitory signals alone is often insufficient for the majority of injured axons to achieve long-distance regeneration, as intrinsic regenerative capacity of mature CNS neurons is also a critical determinant for axon re-growth(Sun et al., 2011). Combinatory strategies that enhance neuronal growth and in the meantime overcome environmental inhibitory cues appear to confer better axonal regeneration and neural repair (Wang et al., 2012). While animal models are instrumental and indispensable to our understanding of CNS responses to injury and investigation of intervention strategies and functional regeneration, in vitro models have been designed to address specific and unique questions due to their accessibilities to experimental manipulations and relatively low cost. For instance, early findings from in vitro culture experiments formed the basis for the concept of CNS myelin-associated inhibitory molecules such as Nogo (Schwab and Thoenen, 1985; Chen et al., 2000; GrandPre et al., 2000). Attempts to identify compounds that overcome the inhibition of CNS myelin and CSPGs on neurite outgrowth in culture have revealed key neuronal signaling components that mediate the inhibitory effects of myelin and CSPGs (Sivasankaran et al., 2004). However, conventional culture system often has to use neuronal cultures at low cell density and for only a short period of time due to technical difficulties in monitoring and quantifying axonal growth. To develop effective CNS regenerative strategies, fast and reliable assessment of axonal growth would be critical not only to identify and select potentially interesting candidate molecules that promote axon extension over inhibitory molecules, but also to rule out poor ones. Equally important are considerations that neurons are highly polarized cells and that damaged axon could be extending far away from the cell body and encountering drastically different microenvironment than that of the soma. As such, signaling events elicited by extrinsic factors are most likely spatially regulated and may have different functional outputs. Over the years, compartmentalized neuronal culture systems have been developed. Now the emergence of microfluidics technology offers many advantages and versatilities for axonal growth and regeneration studies (Figure 1). Figure 1 Schematic illustrations of compartmentalized neuron culture platforms for axon isolation.