Engineered in vitro/in silico models to examine neurite target preference

Research on spinal cord injury (SCI) repair focuses on developing mechanisms to allow neurites to grow past an injury site. In this article, we observe that numerous divergent paths (i.e., spinal roots) are present along the spinal column, and hence guidance strategies must be devised to ensure that regrowing neurites reach viable targets. Therefore, we have engineered an in vitro micropatterned model in which cultured E7 dorsal root ganglia (DRG) explants may enter alternate pathways (?roots?) along a branching micropattern. Alongside this in vitro model, we have developed an in silico simulation that we validate by comparison with independent experiments. We find in both in silico and in vitro models that the probability of a neurite entering a given root decreases exponentially with respect to the number of roots away from the DRG; consequently, the likelihood of neurites reaching a distant root can be vanishingly small. This result represents a starting point for future strategies to optimize the likelihood that neurites will reach appropriate targets in the regenerating nervous system, and provides a new computational tool to evaluate the feasibility and expected success of neurite guidance in complex geometries.