Regeneration across cold preserved peripheral nerve allografts

Abstract

The feasibility of peripheral nerve allograft pretreatment utilizing cold storage (5 degrees C in the University of Wisconsin Cold Storage Solution) or freeze-thawing to prevent rejection was investigated. Regeneration across cold-stored (3 or 5 weeks) or freeze-thawed (FT), 3.0-cm sciatic nerve allografts were compared to fresh auto- and allografts in an inbred rat model. At 16-week post-engraftment, only FT allografts appeared similar to autografts on gross inspection; FT grafts were neither shrunken nor adherent to the surrounding tissue as seen in the other allograft groups. Qualitatively, the pattern of regeneration in the graft segments of the fresh allograft and to a lesser extent of pretreated allografts was inferior to that of autografts as evidenced by a disruption in the perineurium, more extrafascicular axons, smaller and fewer myelinated axons, increased intrafascicular collagen deposition, and the persistence of perineurial cell compartmentation and perivascular infiltrates. Distal to these grafts, the regeneration became more homogenous between groups, although areas of ongoing Wallerian degeneration, new regeneration as well as compartmentation, were more prevalent in fresh and pretreated allografts. Although the number of myelinated fibres was equivalent to autografts, the fibre diameters, the number of large diameter fibres, and the G-ratio were significantly decreased in the allograft groups, which, in part, accounted for the significant decrease in conduction velocity in the 3-week stored and fresh allograft, and the slight decrease in the 5-week stored and FT allograft groups. There was a small return in the Sciatic Function Index towards normal, but no consistent differences between groups were found. Prolonged cold storage and freeze-thawing of nerve allografts resulted in regeneration that was better than fresh allografts, but inferior to autografts. With the concomitant use of host immunosuppression or other immunotherapies, these storage techniques can provide a means of transporting nerve allografts between medical centres and for converting urgent into elective procedures.