Spatial gradients of chemotropic factors from immobilized patterns to guide axonal growth and regeneration
Spatial gradients of chemotropic factors from immobilized patterns to guide axonal growth and regeneration
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DOI：
10.1016/j.biomaterials.2013.08.019
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发表时间：
2013-12-01
期刊：
影响因子：
14
通讯作者：
Ito, Yoshihiro
中科院分区：
文献类型：
--
作者：
Joddar, Binata;Guy, Adam T.;Ito, Yoshihiro
在体外实验中，我们研究了NGF和Sema3A不同的局部浓度梯度对鸡胚DRG外植体和原代神经元轴突生长的影响。利用光刻技术在组织培养覆盖片上制备固定化的二维NGF或Sema3A微图案。两个不同的区域被确定：缓慢，很少或没有变化的趋化因子的浓度；而且很陡，有从低到高的过渡。轴突生长方向定义为近端或远端，近端向固定化NGF/Sema3A浓度较高的方向生长，远端反之。当外植体在陡峭的NGF上播种时，轴突优先向近端生长，而在陡峭的Sema3A下，轴突优先向远端生长。在缓慢的NGF和缓慢的Sema3A上，轴突生长的方向特异性没有差异。植于陡峭NGF上的DRG原代神经元向近端迁移，而植于缓慢NGF上的神经元向各方向迁移。相反，植入陡峭或缓慢Sema3A的神经元没有延长任何轴突。我们的二维固定化因子微模式显示出进一步发展体外神经组织工程研究的前景。(C) 2013 Elsevier Ltd.版权所有。
In this study, we investigated the effect of varying localized concentration gradients of NGF and Sema3A on the axonal outgrowth of embryonic chick DRG explants and primary neurons in vitro. Immobilized 2D NGF or Sema3A micropatterns were produced using photolithography on tissue culture cover slips. Two distinct regions were identified: slow, with little or no change in concentration of chemotropic factor; and steep, with a transition from low to high. The direction of axonal outgrowth was defined as proximal or distal, with proximal growing towards the higher concentration of immobilized NGF/Sema3A and vice versa for distal. Axons grew preferentially in the proximal direction when explants were seeded onto steep NGF, and distally in response to steep Sema3A. On slow NGF, or on slow Sema3A there was no difference in the directional specificity of axonal outgrowth. DRG primary neurons seeded onto steep NGF migrated proximally, whereas neurons seeded onto slow NGF migrated in all directions. Conversely, neurons seeded onto steep or slow Sema3A did not extend any axons. Our 2D immobilized micropatterns of chemotropic factors show promise for further development of in vitro nerve tissue engineering studies. (C) 2013 Elsevier Ltd. All rights reserved.