周围神经组织工程材料的预构

MATERIAL S FABRICATION OF TISSUE ENGINEERED PERIPHERAL NERVE IN VITRO

目的 研究具有雪旺细胞 (SC)活性的组织工程周围神经修复材料的构建方法。方法 1采用干 /湿相转化纺丝法 ,将分子量为 730 k D的聚乳酸 (PL A)制成外径为 2 .3m m、内径为 1.9m m、壁厚 0 .4mm、长4cm、孔隙率为 70 %、孔径为 2 0～ 40 μm的中空纤维管和孔径为 10 0～ 2 0 0 μm、孔隙率为 85 %的 PL A无纺纤维布。2将 SC置入细胞外基质 (ECM)凝胶中 ,观察 SC在 ECM凝胶中的生长。 3SC/ ECM悬液浸入 PL A无纺纤维布中 ,观察 SC在 PL A支架材料中的生长。 4将 SC、SC/ ECM和 SC/ ECM/ PL A置入 PL A中空纤维管中 ,桥接大鼠坐骨神经长 10 mm缺损 ,观察 SC移植后存活情况。结果 1SC在 ECM凝胶中 ,细胞生长良好 ,1天后开始有细胞伸出两极 ,形态开始展开 ,3天后几乎全部细胞恢复成双极梭形状细胞 ,并开始分裂、增殖 ,直到 14天后随着凝胶的溃解 ,SC开始死亡 ,降解成碎片 ;当 ECM凝胶溃解后 ,加入 DMEM培养基 ,SC游出 ,贴壁生长。2 SC/ ECM悬液浸入 PL A无纺纤维布后 ,大部分细胞随 ECM凝胶状态的形成 ,滞留于 PL A纤维网孔中 ,1天后开始伸出两极 ,3天后可见网孔中大量双极梭形细胞 ,部分细胞沿着纤维生长 ,仅少数细胞脱落于培养瓶底贴壁生长。 3SC移植术后2 1天 ,Brd U免疫组织化学染色发现大量

Objective To investigate the methods to fabricate repair materials of tissue engineered pe- ripheral nerve with bioactivity of Schwann cells(SC) .Methods 1The m aterials were m ade by dry- wet spinning process to fabricate PL A hollow fiber canal with external diam eter of2 .3mm,internal diameter of1.9mm,thick- ness of0 .4mm,pore size of2 0 to40 μm,pore ratio of70 % and non- spinning fiber net with pore size of10 0 to2 0 0 μm,pore ratio of85 % . 2 SC were implanted into excellular m atrix(ECM) gel to observe the growth of SC.3 SC/ ECM com plex were implanted into non- spinning PL A fiber net to observe the growth of SC. 4SC,SC/ ECM and SC/ ECM/ PL A were implanted into PL A hollow fiber canal to bridge10 m m defect of ratsciatic nerve.Results 1SC were recovered bipolar shape at1day after implantation,and could be survived14days in ECM gel.2 Af- ter SC/ ECM com plex was im planted into PL A net,m ostof SC were retained in the pore of PL A net with the for- mation of ECM gel.SC could be adhered and grown on PL A fiber. 3Most of SC in ECM gel could be survived to 2 1days after transplantation.Survival cell num bers of SC/ ECM and SC/ ECM/ PL A groups were obviously higher than SC suspension group.Conclusion Non- spinning PL A porous biodegradable m aterials with ECM is benefit for SC to be adhered and grown.