University of Applied Sciences Technikum Wien, Department of Biochemical Engineering, Höchstädtplatz 5, 1200 Vienna, Austria; The Austrian Cluster for Tissue Regeneration, Vienna, Austria.
Department of Pharmaceutical Technology and Biopharmaceutics, University of Vienna, Vienna 1090, Althanstraße 14, Austria.
Acta Biomater. 2014 Jun;10(6):2506-17. doi: 10.1016/j.actbio.2014.02.012. Epub 2014 Feb 12.
Various tissue engineering (TE) approaches are based on silk fibroin (SF) as scaffold material because of its superior mechanical and biological properties compared to other materials. The translation of one-step TE approaches to clinical application has generally failed so far due to the requirement of a prolonged cell seeding step before implantation. Here, we propose that the plant lectin WGA (wheat germ agglutinin), covalently bound to SF, will mediate cell adhesion in a time frame acceptable to be part of a one-step surgical intervention. After the establishment of a modification protocol utilizing carbodiimide chemistry, we examined the attachment of cells, with a special focus on adipose-derived stromal cells (ASC), on WGA-SF compared to pure native SF. After a limited time frame of 20min the attachment of ASCs to WGA-SF showed an increase of about 17-fold, as compared to pure native SF. The lectin-mediated cell adhesion further showed an enhanced resistance to trypsin (as a protease model) and to applied fluid shear stress (mechanical stability). Moreover, we could demonstrate that the adhesion of ASCs on the WGA-SF does not negatively influence proliferation or differentiation potential into the osteogenic lineage. To test for in vitro immune response, the proliferation of peripheral blood mononuclear cells in contact with the WGA-SF was determined, showing no alterations compared to plain SF. All these findings suggest that the WGA modification of SF offers important benefits for translation of SF scaffolds into clinical applications.
各种组织工程 (TE) 方法都基于丝素蛋白 (SF) 作为支架材料，因为它具有比其他材料更优异的机械和生物学性能。迄今为止，由于在植入前需要进行延长的细胞接种步骤，因此一步法 TE 方法的转化通常都失败了。在这里，我们提出，植物凝集素 WGA（麦胚凝集素）与 SF 共价结合，将介导细胞在可接受的时间范围内附着，成为一步手术干预的一部分。在用碳二亚胺化学建立了修饰方案后，我们研究了细胞的附着，特别关注脂肪来源的基质细胞 (ASC)，与纯天然 SF 相比，WGA-SF 的附着。在 20 分钟的有限时间内，与纯天然 SF 相比，ASC 对 WGA-SF 的附着增加了约 17 倍。凝集素介导的细胞附着进一步显示出对胰蛋白酶（作为蛋白酶模型）和施加的流体剪切力（机械稳定性）的增强抗性。此外，我们可以证明 ASC 在 WGA-SF 上的附着不会对成骨谱系的增殖或分化潜力产生负面影响。为了测试体外免疫反应，测定了与 WGA-SF 接触的外周血单核细胞的增殖，与 plain SF 相比没有变化。所有这些发现表明，SF 的 WGA 修饰为 SF 支架转化为临床应用提供了重要的益处。
