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Peripheral nerve injury (PNI) has become one of the common clinical diseases. How to promote the regeneration and function recovery of the damaged peripheral nerve has been the focus of attention in the medical field. Evidence suggests that the longitudinal filling of oriented fibers in nerve guide conduit (NGC) is especially beneficial to the repair of long gap PNI. In this study, polypyrrole (PPy) nanospheres (PNSs) were prepared by the soft-templating method, and mixed with poly (lactic acid) (PLA) to prepare conductive PNSs/PLA NGC, and the optimal ratio of PNSs was 4.5%. PLA and vascular endothelial growth factor (VEGF) as shell, chitosan (CS) and paeoniflorin (PF) as core, oriented coaxial nanofibers were obtained and then filled into PNSs/PLA NGC. The composite NGC has excellent mechanical properties, electrical conductivity, hydrophilic properties, and degradation properties. Besides, the successive release of VEGF and PF can play a synergistic role in promoting nerve regeneration. In vitro experiments showed that the composite NGC was nontoxic and suitable for the adhesion and proliferation of nerve cells. In addition, PNSs combined with electrical stimulation (ES) can significantly promote the differentiation and proliferation of nerve cells, which is conducive to nerve regeneration. These positive results indicate that the composite NGC is a promising candidate in the repair of long gap PNI. [ABSTRACT FROM AUTHOR]