[Comparison of different chemical methods for preparation of acellular nerve scaffold]

Compare the effect of different chemical methods for preparation of acellular nerve scaffold and to provide an effective nerve scaffold for tissue engineering.

Fifteen male SD rats of 2 months old, weighing 200-250 g were selected; the bilateral sciatic nerves were harvested and divided into 3 groups according to preparation methods: group A (normal nerve), group B (Sondell method) and group C (optimal method by Triton X-200, SB-10 and SB-16). The morphology was compared by HE, immunohistochemistry and SEM after dispose; the degrees of decellularization, degrees of demyelination and integrity of the nerve fiber tube were assessed by scoring system.

HE staining: In group A, the cross section of nerve was roundness, the cell nuclei was dark blue and the myelin sheath was reticular structure. In group B, the axon and cell nuclei disappeared and the structure of endoneurium was destroyed. In group C, the axon and cell nuclei disappeared and the endoneurium become animalistic round cavum. The immunohistochemistry staining of Laminin: In group A, the myelin sheath was surrounded by basement membrane with dark blue SC nuclei inside. In group B, the myelin sheath and SC nuclei disappeared and the structure of basement membrane destroyed. In group C, the myelin sheath and SC nuclei disappeared and basement membrane become animalistic round cavum. The immunohistochemistry staining of S-100: In group A, the myelin sheath and SC were brown. In groups B and C, there were no apparent stained myelin sheath. SEM: In group A, the myelin sheath and axon were clear. In group B, the axon and myelin sheath disappeared and basement membrane became animalistic. In group C, the basement membrane was more regular than that of group B. The degrees of acellularization and demyelination of groups B and C were superior to that of group A (P < 0.05), and the degrees of demyelination of group C were superior to that of group B (P < 0.05). The integrity of nerve fiber tube of group C was superior to that of group B (P < 0.05) and similar to that of group A (P > 0.05). The total score was the lowest in group C but the quality was the best.

The effect of decellularization of optimal method was similar to that of traditional Sondell method, but the effect of demyelination and integrity of nerve fiber tube were better than that of traditional Sondell method. And this acellular nerve can be used as a new kind of nerve scaffold material.