The Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, FL 33136-0621, USA.
Biomaterials. 2012 Nov;33(33):8529-39. doi: 10.1016/j.biomaterials.2012.07.028. Epub 2012 Aug 17.
One of the most exciting new avenues of research to repair the injured spinal cord is to combine cells for implantation with scaffolds that protect the cells and release growth factors to improve their survival and promote host axonal regeneration. To realize this goal, we fabricated biodegradable, photocurable gelatin tubes and membranes for exploratory in vitro studies. Detailed methods are described for their fabrication with a high gelatin concentration. Gelatin membranes fabricated in the same way as tubes and photo-co-immobilized with rhBDNF or rhNT-3, with or without Schwann cells (SCs), showed an initial burst of neurotrophin release within 24 h, with release diminishing progressively for 21 days thereafter. SCs attained their typical bipolar conformation on membranes without neurotrophins but adhesion, alignment and proliferation were improved with neurotrophins, particularly rhBDNF. When dorsal root ganglion explants were cultured on membranes containing laminin and fibronectin plus both neurotrophins, neurite outgrowth was lengthier compared to combining one neurotrophin with laminin and fibronectin. Thus, these gelatin membranes allow SC survival and effectively release growth factors and harbor extracellular matrix components to improve cell survival and neurite growth. These scaffolds, based on the combination of cross-linked gelatin technology and incorporation of neurotrophins and extracellular matrix components, are promising candidates for spinal cord repair.
修复受损脊髓的最令人兴奋的新研究途径之一是将用于植入的细胞与保护细胞并释放生长因子以提高细胞存活率和促进宿主轴突再生的支架结合起来。为了实现这一目标,我们制造了可生物降解的、光固化的明胶管和膜,用于探索性的体外研究。详细描述了用高浓度明胶制造它们的方法。以与管相同的方式制造的明胶膜,并与 rhBDNF 或 rhNT-3 光共固定,有或没有施万细胞(SCs),在 24 小时内显示出神经营养因子释放的初始爆发,此后逐渐减少 21 天。SCs 在没有神经营养因子的情况下在膜上获得其典型的双极构象,但粘附、排列和增殖在神经营养因子(特别是 rhBDNF)的存在下得到改善。当背根神经节外植体在含有层粘连蛋白和纤维连接蛋白以及两种神经营养因子的膜上培养时,与将一种神经营养因子与层粘连蛋白和纤维连接蛋白结合相比,神经突生长更长。因此,这些明胶膜允许 SC 存活,并有效地释放生长因子并容纳细胞外基质成分以改善细胞存活和神经突生长。这些支架基于交联明胶技术的结合以及神经营养因子和细胞外基质成分的掺入,是脊髓修复的有前途的候选物。