Objective: Evaluate the biocompatibility of olfactory ensheathing cells (OECs) on the electrospun low-diameter silk fibroin scaffold (LD-SFS). Methods: 400 nm silk fibroin nanofibers were prepared by electrospinning technique and were observed by scanning electron microscope (SEM). The OECs were isolated and purified by the modified differential adherent velocity method. Then, the purified OECs were seed on the poly-L-lysine and electrospun silk fibroin scaffold. The nerve growth factor receptor (NGFR) p75 and glial fibrillary acidic protein (GFAP) were used to identify OECs by immunofluorescence staining. The MTT and flow cytometric assay were used to detect the proliferation and apoptosis effect of OECs on the different scaffolds. Results: The SEM showed that the average diameter of the fibers was about 400 nm and the nanofibers constituted a three-dimensional structure with porous network and smooth surface. The morphology of OECs on the LD-SFS group was similar to that on the poly-L-lysine (PLL) group. In addition, MTT and flow cytometric assay also showed that there was no significant difference between the two scaffolds in the proliferation and apoptosis activity. Conclusion: LD-SFS may serve as an ideal tissue engineering scaffold for the olfactory ensheathing cells.