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新型超薄 PCL 和 PCL/PLA 共混膜作为周围神经导管的体外和体内试验。

In vitro and in vivo testing of novel ultrathin PCL and PCL/PLA blend films as peripheral nerve conduit.

机构信息

Materials Science Centre, Department of Engineering and Physical Sciences, The University of Manchester, Grosvenor Street, Manchester M1 7HS, United Kingdom.

出版信息

J Biomed Mater Res A. 2010 Jun 15;93(4):1470-81. doi: 10.1002/jbm.a.32681.

Abstract

In an attempt to obviate the drawbacks of nerve autograft, ultrathin microporous biodegradable PCL and PCL/PLA films were tested for their compatibility with motor neuron-like NG108-15 cells and primary Schwann cells. Data obtained from MTS colorimetric and DNA fluorimetric assays showed that both cell lines readily attached and proliferated on these materials. Images taken using scanning electron microscope and fluorescence microscope confirmed these observations. Enhanced cell-surface interaction was achieved by pretreating the films in NaOH solution. Importantly, NG108-15 cells could be induced into differentiated phenotype with long, un-branched neurites growing across the surface of the materials. The bipolar spindle-shaped phenotype of Schwann cells was also retained on these scaffolds. Positive immunochemical staining using antibodies against neurofilament for NG108-15 cells and S100 for Schwann cells indicated the expression of these marker proteins. In a small-scaled pilot testing, the performance of PCL conduits in bridging up a 10 mm gap in rat sciatic nerve model was assessed. Immunohistochemical staining showed that regenerated nerve tissue and penetrated Schwann cells have the potential to span the whole length of the conduit in 2 weeks.

摘要

为了避免神经自体移植的缺点,我们测试了超薄微孔可生物降解的聚己内酯(PCL)和聚己内酯/聚乳酸(PCL/PLA)薄膜与类似于运动神经元的 NG108-15 细胞和原代雪旺细胞的相容性。MTS 比色法和 DNA 荧光法获得的数据表明,这两种细胞系都很容易附着并在这些材料上增殖。扫描电子显微镜和荧光显微镜拍摄的图像证实了这些观察结果。通过将薄膜在 NaOH 溶液中预处理,可以实现增强的细胞表面相互作用。重要的是,NG108-15 细胞可以被诱导分化为具有长而无分支突起的形态,这些突起可以穿过材料表面生长。雪旺细胞的双极纺锤形形态也保留在这些支架上。使用针对 NG108-15 细胞的神经丝抗体和针对 Schwann 细胞的 S100 抗体进行的阳性免疫化学染色表明这些标记蛋白的表达。在小规模的初步测试中,评估了 PCL 导管在大鼠坐骨神经模型中桥接 10mm 间隙的性能。免疫组织化学染色显示,再生的神经组织和穿透的雪旺细胞有潜力在 2 周内穿过导管的整个长度。

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