Hoesli Corinne A, Tremblay Catherine, Juneau Pierre-Marc, Boulanger Mariève D, Beland Ariane V, Ling Si Da, Gaillet Bruno, Duchesne Carl, Ruel Jean, Laroche Gaétan, Garnier Alain
Department of Chemical Engineering, Faculty of Engineering, McGill University, Wong Building, 3610 University Street, Montréal, Québec H3A 0C5, Canada.
PROTEO Research Center, Québec, Canada.
ACS Biomater Sci Eng. 2018 Nov 12;4(11):3779-3791. doi: 10.1021/acsbiomaterials.8b00774. Epub 2018 Oct 26.
Surface endothelialization could improve the long-term performance of vascular grafts and stents. We previously demonstrated that aerosol-generated fibronectin-derived peptide micropatterns consisting of GRGDS spots over a WQPPRARI background increase endothelial cell yields in static cultures. We developed a novel fluorophore-tagged RGD peptide (RGD-TAMRA) to visualize cell-surface interactions in real-time. Here, we studied the dynamics of endothelial cell response to laminar flow on these peptide-functionalized surfaces. Endothelial cells were exposed to 22 dyn/cm wall shear stress while acquiring time-lapse images. Cell surface coverage and cell alignment were quantified by undecimated wavelet transform multivariate image analysis. Similar to gelatin-coated surfaces, surfaces with uniform RGD-TAMRA distribution led to cell retention and rapid cell alignment (∼63% of the final cell alignment was reached within 1.5 h), contrary to the micropatterned surfaces. The RGD-TAMRA peptide is a promising candidate for endothelial cell retention under flow, and the spray-based micropatterned surfaces are more promising for static cultures.
表面内皮化可改善血管移植物和支架的长期性能。我们之前证明,在WQPPRARI背景上由GRGDS斑点组成的气溶胶生成的纤连蛋白衍生肽微图案可提高静态培养中的内皮细胞产量。我们开发了一种新型的荧光团标记的RGD肽(RGD-TAMRA)以实时可视化细胞-表面相互作用。在此,我们研究了内皮细胞对这些肽功能化表面上的层流的反应动力学。在内皮细胞暴露于22达因/平方厘米的壁面剪应力的同时采集延时图像。通过未抽取小波变换多变量图像分析对细胞表面覆盖率和细胞排列进行量化。与明胶包被的表面类似,具有均匀RGD-TAMRA分布的表面导致细胞滞留和快速细胞排列(在1.5小时内达到最终细胞排列的约63%),这与微图案化表面相反。RGD-TAMRA肽是流动条件下内皮细胞滞留的一个有前景的候选物,而基于喷雾的微图案化表面在静态培养中更具前景。