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Transplantation of NT-3 delivery acellular optic nerve scaffold to improve motor function in transected rat spinal cord

Transplantation of NT-3 delivery acellular optic nerve scaff...

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中国神经科学学会第十二届全国学术会议

作者： Ge Li Jia-Hui Sun Yuan-Shan Zeng Key Laboratory for Stem Cells and Tissue Engineering (Sun Yat-sen University) Ministry of Education Department of Histology and Embryology Zhongshan School of Medicine Sun Yat-sen University

Objective To generate a bioactive scaffold with line-shaped tubes in vitro and investigate whether the scaffold transplanted into the injury site could promote nerve fiber regeneration and functional recovery in rat spinal cord transected completely. Methods By using aseptic decellular, DNAase hydrolysis and freeze-drying techniques, natural optic nerve was removed all cell component and coated with neurotrophin-3（NT-3）/fibroin complex at the surface to fabricate a NT-3 delivery acellular optic nerve scaffold. Then, dorsal root ganglion（DRG） was seeded on the surface of NT-3 delivery acellular optic nerve scaffold in vitro to observe the effect of the scaffold on DRG neurite growth, and the scaffold were transplanted into the injury site of spinal cord injury of rat. Results Fresh normal optic nerve of longitudinal section was stained by HE showing high-density tubes with optic nerve fibers. By using aseptic decellular technique and freeze-drying technique, the NT-3 delivery acellular optic nerve scaffold still maintained its natural 3 dimension（3 D） structure of extracellular matrix. Circle-shaped pipelines made of perineurium-like extracellular matrix in optic nerve of transverse section could be obseved and there were line-shaped tubes with interconnections in optic nerve of longitudinal section after all cell component was removed. These results were further conformed under a scanning electron microscope. The present study proved that cell component had been removed by the results below;*** nuclei were invisible by Hoechst33342 staining. 2. DNA electrophoresis showed that residual DNA fragments were less than 200 bp. 3. DNA quantification showed ＜ 50 ng per mg scaffold dry weight. Therefore, cells seeded in the scaffold could grow without nucleic acid residue effect. Then, after DRG was seeded on the scaffold in vitro, we found that DRG derived NF200 positive neurites could grow a long distance in the surface of the scaffold. Furthermore, two months later aft

关键词： Neutrophin-3 delivery Acellular optic nerve Neurite growth nerve fiber regeneration Spinal cord injury

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