Cryopreservation of peripheral nerve grafts

The utilization of viable biological nerve graft substitutes and nerve allografts raises the problem of nerve storage. To clarify this, rat sciatic nerve segments were harvested and stored in Dulbecco's modified eagle medium. The segments were divided into three groups. In the first group, no cryoprotectant was added, whereas the second had 10% dimethyl sulfoxide (DMSO) added as cryoprotectant. These two groups of nerve segments were subjected to controlled freezing. In a third group, segments were frozen uncontrolled in liquid nitrogen (-196 degrees C). All nerves were replanted orthotopically. Fresh conventional autografts (fourth group) served as control group. Histologically, freezing did not affect the structural elements such as basal lamina tubes and perineurial tissue. Morphometrically, all cryopreserved grafts had significantly reduced axon counts and less myelinization than did controls. Cryoprotected nerves (group 2) showed no different morphometric parameters compared with the group without DMSO (group 1). Controlled freezing was superior to uncontrolled freezing (group 3). Impaired regeneration was attributed mainly to delayed Wallerian degeneration and slower revascularization. Moreover, decreased survival of resident Schwann cells in the graft may impair regeneration due to the lack of neurotrophic, neurotropic, and attachment factors in early regeneration. Grafts subjected to controlled freezing support axonal regeneration to a certain extent, but further studies are required to assess various cooling patterns, cryoprotectants, and graft revascularization.