Tissue engineering of blood vessels: characterization of smooth-muscle cells for culturing on collagen-and-elastin-based scaffolds
Tissue engineering of blood vessels: characterization of smooth-muscle cells for culturing on collagen-and-elastin-based scaffolds
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DOI：
10.1042/ba20030105
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发表时间：
2004-04-01
影响因子：
2.8
通讯作者：
Vermes, I
中科院分区：
文献类型：
--
作者：
Buijtenhuijs, P;Buttafoco, L;Vermes, I
组织工程学提供了开发模拟自然动脉形态的血管支架的机会。我们已经开发了一种由胶原纤维和弹性蛋白散布在一起的多孔三维支架。将不溶性I型胶原和不溶性弹性蛋白悬浮液冷冻干燥，得到支架。为了提高得到的矩阵的稳定性，用两种不同的方法对它们进行了交联。将单独或与二胺结合的水溶性碳二亚胺用于此目的：获得零长度或非零长度的交联键。通过监测交联前后支架的热行为和自由氨基含量，验证了交联剂的存在。培养不同时间的血管平滑肌细胞(SMC)，观察其生长增殖能力。从人脐静脉和隐静脉分离培养的SMC，用特异性单抗(MAb)染色鉴定其纯度。用抗肌动蛋白和波形蛋白的单抗对培养细胞进行鉴定。14天后，未交联型支架上可见一层融合的SMC。对于交联样品，用两种不同方法交联的支架在细胞附着和增殖方面没有观察到差异。培养在支架上的细胞用抗(α-平滑肌肌动蛋白)单抗鉴定。SMC的取向与胶原纤维和弹性蛋白纤维的取向相似。这样，就有可能开发出与天然血管形态特征相似的管状多孔支架。
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-crosslinked 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.