丝胶蛋白对仿生丝素蛋白神经导管的改性研究  

The modification research of bionic silk fibroin nerve guidance conduits by silk sericin

作　　者：饶建伟[1] 叶舟[1] 占蓓蕾[1] 全大萍[2] 许扬滨[3] 

机构地区：[1]衢州市人民医院骨科,浙江省衢州市324000 [2]中山大学化学与化学工程学院医用高分子降解实验室 [3]中山大学附属第一医院整形修复外科

出　　处：《中华显微外科杂志》2016年第3期251-257,共7页Chinese Journal of Microsurgery

基　　金：广东省科技计划项目（2014A020212479）;广州市科技计划项目（201300000174）

摘　　要：目的探索丝胶（SS）蛋白对仿生多通道丝素（sF）蛋白神经导管（NCs）的改性作用。方法在丝素蛋白水溶液中按比例加入丝胶蛋白，通过循序冷冻工艺制备丝胶／丝素共混神经导管（SS／SF-NCs），扫描电镜（SEM）观测其通道结构，X线衍射及红外光谱检测其内部分子结构，测量其孔隙率及机械性能，MTY实验量化分析PCI2细胞与SS／SF-NCs共培养后的细胞活性，PCI2细胞被用于检测从SS／SF—NCs中释放的NGF的细胞活性。结果SEM结果显示与SF-NCs相比，SS／SF-NCs具有周围神经束仿生结构的线性化导向的多通道，通道分布均匀呈片状纵向均匀排列，且通道微观结构上发生很大变化，机械性能得到很大提升，通道的壁间距、孔隙率、机械强度随着循序冷冻温度、丝胶比例的不同而变化。MTF试验结果显示SS／SF—NCs的PCI2细胞的活性明显好于对照组（P〈0．05），从新型SS／SF-NCs中释放NGF释放时间长达4周并保持生物活性。结论通过丝胶蛋白的改性作用，新型SS／SF．NCs具有周围神经束的高度仿生结构，且具有更加优良的机械性能，可以作为人工神经导管的另一选择。Objective To explore the modification of bionic silk fibroin nerve conduits （SF-NCs） by silk sericin. Methods The innovative SS/SF blended-NCs was fabricated by a vertical sequential cooling thermal in- duced phase separation （TIPS） processing with SF solution added sericin in proportion, its morphology was observed by Scanning electron microscopy （SEM）, X-ray diffraction （XRD） and infrared spectroscopy （FT1R） were used to detect its internal molecular structure. MYr assay was used to quantitatively analyzed the PC12 cells viability co-cultured with the innovative SS/SF-NCs, SEM was used to observe the adhesion and morphology of PC12 cells seeded into the innovative SS/SF, PC12 cells were used to assess the NGF bioactivity released from the SS/SF. Results The SEM results showed that the new fabricated SS/SF-NCs had linearly oriented lamellar-like multiple-channel which distribut- ed evenly, got great changes on the channel microstructure and their mechanical properties had been greatly improved, compared to SF-NCs. The XRD and FTIR results showed that the SS/SF-NCs had the similar internal molecular struc- ture with natural silk. The spaces between parallel lamellar-like channels, porosities and compressive strengths of the SS/SF-NCs decreased with decreasing Sequential freezing temperature. MTY assay results showed that the viability of PC12 cell was better than the control group （P 〈 0.05）. The SEM observation indicated that PC12 cells showed good adhesion and differentiation with neuritis outgrowth during the period of co-culture with the SS/SF-NCs. NGF release from the innovative SS/SF-NCs was prolonged over 4 weeks, and remained bioactive. Conclusion The new fabri- cated SS/SF-NCs modified though silk sericin, which was highly bionic the structure of peripheral nerve fasciculus, had excellent mechanical properties and could be used as another alternative of artificial nerve conduits.

关 键 词：神经导管 丝胶/丝素共混 仿生 改性 周围神经 

分 类 号：R688[医药卫生—骨科学]