Wacker Max, Riedel Jan, Walles Heike, Scherner Maximilian, Awad George, Varghese Sam, Schürlein Sebastian, Garke Bernd, Veluswamy Priya, Wippermann Jens, Hülsmann Jörn
Department of Cardiothoracic Surgery, University Hospital Magdeburg, 39112 Magdeburg, Germany.
Core Facility Tissue Engineering, Otto-Von-Guericke University Magdeburg, 39106 Magdeburg, Germany.
Nanomaterials (Basel). 2021 Jul 29;11(8):1952. doi: 10.3390/nano11081952.
In this study, we contrast the impacts of surface coating bacterial nanocellulose small-diameter vascular grafts (BNC-SDVGs) with human albumin, fibronectin, or heparin-chitosan upon endothelialization with human saphenous vein endothelial cells (VEC) or endothelial progenitor cells (EPC) in vitro. In one scenario, coated grafts were cut into 2D circular patches for static colonization of a defined inner surface area; in another scenario, they were mounted on a customized bioreactor and subsequently perfused for cell seeding. We evaluated the colonization by emerging metabolic activity and the preservation of endothelial functionality by water soluble tetrazolium salts (WST-1), acetylated low-density lipoprotein (AcLDL) uptake assays, and immune fluorescence staining. Uncoated BNC scaffolds served as controls. The fibronectin coating significantly promoted adhesion and growth of VECs and EPCs, while albumin only promoted adhesion of VECs, but here, the cells were functionally impaired as indicated by missing AcLDL uptake. The heparin-chitosan coating led to significantly improved adhesion of EPCs, but not VECs. In summary, both fibronectin and heparin-chitosan coatings could beneficially impact the endothelialization of BNC-SDVGs and might therefore represent promising approaches to help improve the longevity and reduce the thrombogenicity of BNC-SDVGs in the future.
在本研究中,我们对比了用人类白蛋白、纤连蛋白或肝素 - 壳聚糖对细菌纳米纤维素小直径血管移植物(BNC - SDVGs)进行表面涂层后,其对人隐静脉内皮细胞(VEC)或内皮祖细胞(EPC)在体外内皮化的影响。在一种情况下,将涂层移植物切成二维圆形贴片,用于在确定的内表面积上进行静态定植;在另一种情况下,将它们安装在定制的生物反应器上,随后进行灌注以接种细胞。我们通过新出现的代谢活性评估定植情况,并通过水溶性四氮唑盐(WST - 1)、乙酰化低密度脂蛋白(AcLDL)摄取试验和免疫荧光染色来评估内皮功能的保留情况。未涂层的BNC支架用作对照。纤连蛋白涂层显著促进了VEC和EPC的黏附与生长,而白蛋白仅促进了VEC的黏附,但此处细胞在功能上受损,表现为缺乏AcLDL摄取。肝素 - 壳聚糖涂层显著改善了EPC的黏附,但对VEC没有作用。总之,纤连蛋白和肝素 - 壳聚糖涂层都可能对BNC - SDVGs的内皮化产生有益影响,因此可能代表了未来有助于提高BNC - SDVGs的使用寿命并降低其血栓形成性的有前景的方法。