Guided cell adhesion and outgrowth in peptide-modified channels for neural tissue engineering
Guided cell adhesion and outgrowth in peptide-modified channels for neural tissue engineering
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DOI：
10.1016/j.biomaterials.2004.05.012
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发表时间：
2005-05-01
期刊：
影响因子：
14
通讯作者：
Shoichet, MS
中科院分区：
文献类型：
--
作者：
Yu, TT;Shoichet, MS
创建了具有明确几何形状的水凝胶支架，并用水溶液中的层粘连蛋白衍生肽进行修饰，从而保持支架的几何形状，同时引入增强细胞粘附和神经突生长的生物活性肽。通过将纤维模板技术与肽修饰相结合来创建纵向通道，我们能够合成引导原代神经元的细胞粘附和神经突生长的支架。支架被设计为具有许多纵向通道，平均通道直径为 196+/-6μm，以最终促进再生电缆的束状收缩，压缩模量为 192+/-8kPa，以匹配软神经组织的模量。甲基丙烯酸 2-羟乙酯 (HEMA) 与甲基丙烯酸 2-氨基乙酯 (AEMA) 支架的共聚提供了伯胺基团，使用磺基-(N-马来酰亚胺甲基)环己烷-1-羧酸酯 (磺基-SMCC) 交联剂将两个硫氢基封端的层粘连蛋白衍生寡肽 CDPGYIGSR 和 CQAASIKVAV 共价结合到伯胺基团上。使用茚三酮法测量支架上肽的浓度为106+/-4 mumol/cm(2)，使用BCA蛋白质测定法测量支架上肽的浓度为92+/-9 mumol/cm(2)。肽修饰的 P(HEMA-co-AEMA) 支架在水性条件下很容易制造，与非肽修饰的对照相比，具有高度可重复性、轮廓清晰、增强的神经细胞粘附和引导原代鸡背根神经节神经元的神经突生长。 AEMA 与 HEMA 的共聚可以扩展到其他自由基聚合单体，并且是有利的，因为它有利于水溶液中的支架修饰，从而避免使用可能具有细胞毒性并经常破坏支架几何形状的有机溶剂。本文描述的明确的化学和物理刺激的组合提供了用于体外和体内引导再生的手段。 (C) 2004 Elsevier Ltd. 保留所有权利。
A hydrogel scaffold of well-defined geometry was created and modified with laminin-derived peptides in an aqueous solution, thereby maintaining the geometry of the scaffold while introducing bioactive peptides that enhance cell adhesion and neurite outgrowth. By combining a fiber templating technique to create longitudinal channels with peptide modification, we were able to synthesize a scaffold that guided cell adhesion and neurite outgrowth of primary neurons. Scaffolds were designed to have numerous longitudinally oriented channels with an average channel diameter of 196+/-6 mum to ultimately promote fasciculation of regenerating cables and a compressive modulus of 192+/-8 kPa to match the modulus of the soft nerve tissue. Copolymerization of 2-hydroxylethyl methacrylate (HEMA) with 2-aminoethyl methacrylate (AEMA) scaffolds, provided primary amine groups to which two sulfhydryl terminated, laminin-derived oligopeptides, CDPGYIGSR and CQAASIKVAV, were covalently bound using the sulfo-(N-maleimidomethyl)cyclohexane-1-carboxylate (sulfo-SMCC) crosslinking agent. The concentration of peptides on the scaffolds was measured at 106+/-4 mumol/cm(2) using the ninhydrin method and 92+/-9 mumol/cm(2) using the BCA protein assay. The peptide modified P(HEMA-co-AEMA) scaffolds were easily fabricated in aqueous conditions, highly reproducible, well-defined, and enhanced neural cell adhesion and guided neurite outgrowth of primary chick dorsal root ganglia neurons relative to non-peptide-modified controls. The copolymerization of AEMA with HEMA can be extended to other radically polymerized monomers and is advantageous as it facilitates scaffold modification in aqueous solutions thereby obviating the use of organic solvents which can be cytotoxic and often disrupt scaffold geometry. The combination of well-defined chemical and physical stimuli described herein provides a means for guided regeneration both in vitro and in vivo. (C) 2004 Elsevier Ltd. All rights reserved.