We used predegenerated acellular grafts to bridge proximal and distal stumps of transected nerves and studied how the duration of predegeneration might affect axonal regeneration. Predegenerated acellular grafts were prepared by transecting the tibial nerve of donor rats and, after a period of degeneration, freeze-thawing a 40-mm long segment of the distal stump. Five degeneration periods were used: 0 days (for fresh grafts), 3 days, 1 week, 4 weeks, and 8 weeks. Fresh cellular grafts not treated with freeze-thawing were also used for comparison. Each graft was then transplanted to an isogeneic recipient rat, in which it was used to bridge the proximal stump of the transected left tibial nerve and the distal stamp of the transected right tibial nerve. Six weeks were allowed for the regeneration of axons in all grafts. The regeneration was then assessed by studying transverse sections of the grafts, to determine the maximum length that the axons had regenerated, and the packing density of axons (percentage of sampled areas occupied by axons). The results show that axons had grown to the maximum length in the 4-week predegenerated grafts, and had the highest packing density in the 1-week predegenerated grafts. Regeneration in the fresh acellular (0-day predegenerated) and 8-week predegenerated grafts, especially the latter, was poor. We examine the results with reference to time-dependent events of Wallerian degeneration and propose that there are beneficial effects of multiple factors on the grafts during the first 4 weeks of predegeneration, causing a slow but significant improvement in their capability to support axonal growth. The subsequent rapid deterioration of such capability may be related to structural changes in the extracellular scaffold.