Objective To study the feasibility of microsurgical technique to denervate sympathetic of femoral artery in rabbit, providing a reliable animal experimental model for further study of the mechanism of neuralization in bone tissue engineering. Methods From July, 2014 to July, 2015, 21 New Zealand white rabbits were divided into 4 groups randomly: the control group (n = 3), the 4 weeks group (n = 6), the 8 weeks group (n = 6) and the 12 weeks group (n = 6). Bilateral femoral arteries of the 21 rabbits were exposed. Adventitia of femoral arteries in 3 test groups were removed for about 2cm by microsurgical technique, whereas adventitia of the control group remained intact without any treatment. The arteries samples were collected at 4 weeks, 8 weeks and 12 weeks after treatment. The structure of vascular were indicated by hematoxylin-eosin (HE) staining, and the distribution and volume of the sympathetic fibers were evidenced by glyoxylic acid staining and the expression of tyrosine hydroxylase (TH), the marked protein of sympathetic. Results The adventitia of 3 test groups were invisible or lost most of it while the control group remained intact shown by HE staining. For glyoxylic acid staining, the fluorescence intensity value of the control group, 4 weeks group, 8 weeks and 12 weeks were 0.08124±0.00260, 0.02920±0.00206, 0.02661±0.00233, 0.03094±0.00211, respectively (n = 6). The distribution and fluorescence intensity of sympathetic nerve were both significantly reduced in test groups compared to the control group (P 0.05). Semiquantitative analysis of the expression of TH was 0.8626±0.03519, 0.3631±0.03019, 0.3964±0.02239, 0.3487±0.02356 respectively, which showed the same tendency as glyoxylic acid staining test. Conclusion Microsurgical technique is promising as an ideal method for the local denervation of sympathetic nerve from artery system as it can significantly reduce sympathetic fibers on adventitia without regeneration during the experimental period. Key words: Tissue engineering bone; Neuralization; Adventitia; Sympathetic nerve; Microsurgical technigue