Acellular allograft (ACA) improves the repair and reconstruction of long peripheral nerve defects. ω-3 Polyunsaturated fatty acids (PUFAs) carry a neuroprotective potential, and their effects on ACA bridging were elucidated. Thirty rats with long gap sciatic nerve defects (15 mm long) were randomly divided into three groups (
n
=
10
): ACA, ACA + PUFAs, and autograft (AU). Limb condition, wet weight of tibialis anterior muscle (TAM), nerve electrophysiology, S-100, horseradish peroxidase (HRP), and percentage of splenic CD4+ and CD8 + T-lymphocytes were evaluated for 12 weeks after the operation. Rats in the AU and ACA + PUFA groups showed superior condition in affected limbs compared to the ACA group. At 12 wk after surgery, the wet weight of TAM in the ACA + PUFA group was higher than that in the ACA group (
0.4519
±
0.1185
vs.
0.3049
±
0.1272
;
P
<
0.01
) but lower than that in the AU group (
0.4519
±
0.1185
,
0.5628
±
0.0092
;
P
<
0.05
). In all the three groups, sole irritation elicited withdrawal reflex, and S-100 staining was detected in plantar skin. Moreover, horseradish peroxidase staining was overt in both the ventral horn and dorsal root ganglion of the spinal cord. Nerve conduction velocity (m/s), amplitude of action potential (mV), or somatosensory evoked potentials in ACA + PUFAs (
28.81
±
1.04
,
2.20
±
0.27
,
6.98
±
0.29
) were significantly different from that in the AU (
35.71
±
1.28
,
1.81
±
0.19
,
8.15
±
0.52
;
P
<
0.05
) and ACA (
20.03
±
1.94
,
2.95
±
0.36
,
5.22
±
0.53
;
P
<
0.01
) groups. The percentages of splenic CD4+ and CD8+ cells were similar among the three groups. Omega-3 PUFAs improve the bridging effect of ACA on long gap peripheral nerve defects by promoting neuroprotection without arousing an immune response.