In situ endothelialisation potential of a biofunctionalised nanocomposite biomaterial-based small diameter bypass graft
In situ endothelialisation potential of a biofunctionalised nanocomposite biomaterial-based small diameter bypass graft
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DOI：
10.3233/bme-2009-0597
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发表时间：
2009-01-01
影响因子：
1
通讯作者：
Seifalian, Alexander M.
中科院分区：
文献类型：
--
作者：
de Mel, Achala;Punshon, Geoffrey;Seifalian, Alexander M.
与心血管移植相关的内皮功能障碍或缺乏内皮是移植物失败的主要原因，移植物失败与血栓形成和相关并发症有关。本研究旨在(1)使一种基于小直径血管移植物的纳米复合生物材料——多面体低聚硅氧烷修饰的聚碳酸酯脲-氨基甲酸酯（POSS-PCU）具有生物功能；(2)用从外周血中提取的含有EPC的单核细胞诱导内皮化。(1)纳米复合聚合物的生物功能化：利用fmoc化学合成了具有生物活性的RGD肽，它是细胞外基质成分纤维连接蛋白的功能域。在生物材料上附着月桂酸疏水“尾巴”以优化RGD取向。该肽与POSS-PCU交联。(2)从健康成人志愿者外周血中提取祖细胞，静培养于多孔生物功能化纳米复合聚合物上。细胞也被引入移植物连接的电路中，并在细胞引入72小时后引入非静态脉动流条件。用Alamar Blue法检测细胞生长程度。通过扫描电镜和内皮细胞标记物CD34、CD31和eNOS的免疫染色证实内皮化。水接触角测量表明，生物功能化提高了纳米复合聚合物的亲水性。Alamar蓝表明生物功能化纳米复合材料上存在更多的细胞，并且这种细胞活力的相对增加是RGD特有的，正如RAD肽所证实的那样。扫描电镜提供了内皮细胞形态的证据，并与内皮细胞标记物免疫染色证实了这一点。基于生物功能纳米复合聚合物的小直径旁路移植术显示了从外周血中提取的细胞相对快速内皮化的潜力。
Endothelial dysfunction or the lack of an endothelium associated with cardiovascular grafts is a major cause of graft failure which is linked to thrombosis and related complications. This study was aimed to (1) biofunctionalise a nanocomposite biomaterial, Polyhedral Oligomeric silsesquioxane modified polycarbonate urea-urethane (POSS-PCU), based small diameter vascular graft and to (2) induce endothelialisation with EPC containing monocytes, which were extracted from peripheral blood. (1) Biofunctionalisation of the nanocomposite polymer: bioactive RGD peptide, which is a functional domain of an extracellular matrix component, fibronectin, was synthesised using fmoc chemistry. A lauric acid hydrophobic "tail" was attached to optimise the RGD orientation on the biomaterial. The peptide was cross linked to POSS-PCU. The presence of RGD on the nanocomposite was tested with water contact angle measurements and specificity tests were carried out with the peptide RAD (2) Progenitor cells were extracted from peripheral blood of adult healthy volunteers and cultured on porous biofunctionalised nanocomposite polymer under static conditions. Cells were also introduced to a circuit to which the grafts are connected and non static pulsatile flow conditions were introduced after 72 h following cell introduction. The degree of cell growth was tested with Alamar Blue assay. Endothelialisation was confirmed with SEM and by immunostaining for endothelial cell markers, CD34, CD31 and eNOS. Water contact angle measurement indicated that biofunctionalisation had increased hydrophilicity of the nanocomposite polymer. Alamar blue indicated a greater presence of cells on biofunctionalised nanocomposite and this relative increase in cell viability was specific to RGD as confirmed with RAD peptides. SEM provided evidence for endothelial cell morphology and this was confirmed with endothelial cell markers with immunostaining. Biofunctionalised nanocomposite polymer-based small diameter bypass graft demonstrated the potential for relatively rapid endothelialisation from cells extracted from peripheral blood.