Study In Immunologic Rejection Of The Bridging Complex Of Variant Nerve In Repairing The Defect Of Rabitt's Sciatic Nerve

Objective:.In this experiment the compound nerve was made by fetus rabbit Schwann cells into acellular nerve allograft.It was applied to repair defects of rabbit sciatic nerve to observe immunological rejection and confirm the antigenicity of the nerve complex.Methods: 1 The building of compound nerve maded by fetus rabbit Schwann cells into acellular nerve allograft.1.1 Preparation of the Schwann cell cultures: Execute the adopted pregnancy New Zealand white rabbit and taken out the fetus rabbits of 28 days conceptus age. Under operation microscope, both sides of sciatic nerves were achieved and the external connective tissue of the nerve segments was stripped.We applied enzyme digestion (trypsin / collagenase) to culture Scs. In order to remove and hold-back fibroblast, the double 30min differential adhesion culture were used in the experiment. We even added NGF (40ng/l) to FBS to promote SCs growing. Fetus rabbit Schwann cells cultured in vitro was identified by S-100 immune histochemistry. 1.2 Preparation of Extracted Nerve Grafts (eNG): Two sides of the adult New Zealand white rabbit sciatic nerves were used to prepare the eNG. Before the extraction, shear the adipose tissue and part of the epineurium with the operation microscope. Then they were dipped in 4oC distilled water immediately, Soaked for 6hs. After that, immerge them into 3% Triton X-100 solution. 1.3 Schwann cells and acellular nerve allograft were cultured together in vitro: Cell suspension containing 90% of SCs were micro-injected under abinocular microscope in the middle of the extracted nerve graft by a stainless needle, glass-cylinder 100ul microsyringe. 2 Let the acellular nerve allograft transplanted with cultured fetus rabbit Schwann cells bridge the sciatic nerve. We taked 10 major healthy New Zealand rabbits and hocused them by su mian xin(0.1ml/kg) through the vein at auris edge. Sciatic nerve was unfeld through the arc cut on the nearside hip and thigh,and then made the 20mm lack. Take the allograft above to bridge the 20mm defect with 9/0 line of no trauma,and then sewed up the muscle and skin.3 Take out the vein blood and examine the T lymphocyte 5 days befor operation.4 Observe their psychosis,bite and sup,and cicatrisation after operation.5 Tectology espial:observe the growth of Schwann cells 5 days after operation.6 Take out the vein blood and examine the T lymphocyte 5 days after operation.7 Statistical analysis :Use analysis of variance.Results: All rabbits′s mental state and meatand drink weregood.They were lame and there was no infection in cuts.Schwann cells interconnected and aligned in microtubulein reactivate nerve fiber and they formed cell-chain like Büngner's band. Befor and after transplant CD4+ and CD8+T lymph-cell′s ratio in peripheral blood had no change,and CD4+/CD8+ also had no change.Conclusions: The compound nerve maded by fetus rabbit Schwann into acellular nerve allograft has all-right histocompatibility and has no rejection in host and didn't cause general immune reaction. So the compound nerve maded by fetus rabbit Schwann cells into acellular nerve allograft is possible to become an ideal tissue engineered nerve.