Following peripheral nerve injury, rapid and spatially oriented axonal outgrowth from the proximal nerve stump is required for successful tissue regeneration. Regenerative strategies such as introducing fiber bundles into the nerve guidance conduits improve the directional growth of neurons and Schwann cells. Recently, it has been proposed that fiber profiling increases cell alignment and could accelerate neuronal growth. Here, we evaluate the impact of fiber profiling on the extent of neurite outgrowth in vitro as compared to non-profiled round fibers. We developed novel profiled trilobal poly(lactic acid) (PLA) fibers and systematically tested their potency to support nerve regeneration in vitro. The profiled fibers did not improve neurite outgrowth as compared to the round fibers. Instead, we show that growing neurites are merely guided by the type and quantity of proteins adsorbed on the polymer surface. Together this data has significant implications for in vivo experiments focusing on directional regrowth of severed axons across lesion sites during peripheral nerve regeneration.