Buijtenhuijs Paula, Buttafoco Laura, Poot Andre A, Daamen Willeke F, van Kuppevelt Toin H, Dijkstra Pieter J, de Vos Rob A I, Sterk Lotus M Th, Geelkerken Bob R H, Feijen Jan, Vermes Istvan
Polymer Chemistry and Biomaterials Group, Department of Science and Technology, Faculty of Science and Technology, University of Twente, P.O. Box 217, 7500 AE Enschede, The Netherlands.
Biotechnol Appl Biochem. 2004 Apr;39(Pt 2):141-9. doi: 10.1042/BA20030105.
Tissue engineering offers the opportunity to develop vascular scaffolds that mimic the morphology of natural arteries. We have developed a porous three-dimensional scaffold consisting of fibres of collagen and elastin interspersed together. Scaffolds were obtained by freeze-drying a suspension of insoluble type I collagen and insoluble elastin. In order to improve the stability of the obtained matrices, they were cross-linked by two different methods. A water-soluble carbodi-imide, alone or in combination with a diamine, was used for this purpose: zero- or non-zero-length cross-links were obtained. The occurrence of cross-linking was verified by monitoring the thermal behaviour and the free-amino-group contents of the scaffolds before and after cross-linking. Smooth-muscle cells (SMCs) were cultured for different periods of time and their ability to grow and proliferate was investigated. SMCs were isolated from human umbilical and saphenous veins, and the purity of the cultures obtained was verified by staining with a specific monoclonal antibody (mAb). Cultured cells were also identified by mAbs against muscle actin and vimentin. After 14 days, a confluent layer of SMCs was obtained on non-cross-linked scaffolds. As for the cross-linked samples, no differences in cell attachment and proliferation were observed between scaffolds cross-linked using the two different methods. Cells cultured on the scaffolds were identified with an anti-(alpha-smooth-muscle actin) mAb. The orientation of SMCs resembled that of the fibres of collagen and elastin. In this way, it may be possible to develop tubular porous scaffolds resembling the morphological characteristics of native blood vessels.
组织工程为开发模仿天然动脉形态的血管支架提供了契机。我们研发了一种由交织在一起的胶原蛋白纤维和弹性蛋白纤维组成的多孔三维支架。通过冷冻干燥不溶性I型胶原蛋白和不溶性弹性蛋白的悬浮液来获得支架。为了提高所得基质的稳定性，采用两种不同方法对其进行交联。为此使用了水溶性碳二亚胺，单独使用或与二胺联合使用：获得了零长度或非零长度交联。通过监测交联前后支架的热行为和游离氨基含量来验证交联的发生情况。将平滑肌细胞（SMC）培养不同时间段，并研究其生长和增殖能力。从人脐静脉和大隐静脉中分离出SMC，并用特异性单克隆抗体（mAb）染色来验证所得培养物的纯度。还用抗肌肉肌动蛋白和波形蛋白的mAb对培养细胞进行鉴定。14天后，在未交联的支架上获得了一层汇合的SMC。至于交联样品，使用两种不同方法交联的支架在细胞附着和增殖方面未观察到差异。用抗（α-平滑肌肌动蛋白）mAb对在支架上培养的细胞进行鉴定。SMC的取向类似于胶原蛋白纤维和弹性蛋白纤维的取向。通过这种方式，有可能开发出类似于天然血管形态特征的管状多孔支架。
