Zhang Xiaohui, Baughman Cassandra B, Kaplan David L
Department of Chemical and Biological Engineering and Biomedical Engineering, Tufts University, 4 Colby Street, Medford, MA 02155, USA.
Biomaterials. 2008 May;29(14):2217-27. doi: 10.1016/j.biomaterials.2008.01.022. Epub 2008 Feb 14.
Human aortic endothelial (HAEC) and human coronary artery smooth muscle cell (HCASMC) responses on electrospun silk fibroin scaffolds were studied to evaluate potential for vascular tissue engineering. Cell proliferation studies supported the utility of this biomaterial matrix by both HAECs and HCASMCs. Alignment and elongation of HCASMCs on random non-woven nanofibrous silk scaffolds was observed within 5 days after seeding based on SEM and confocal microscopy. Short cord-like structures formed from HAECs on the scaffolds by day 4, and a complex interconnecting network of capillary tubes with identifiable lumens was demonstrated by day 7. The preservation of cell phenotype on the silk fibroin scaffolds was confirmed by the presence of cell-specific markers, including CD146, VE-cadherin, PECAM-1 and vWF for HAECs, and SM-MHC2 and SM-actin for HCASMCs at both protein and transcription levels using immunocytochemistry and real-time RT-PCR, respectively. Formation of ECM was also demonstrated for the HCASMCs, based on the quantification of collagen type I expression at protein and transcription levels. The results indicate a favorable interaction between vascular cells and electrospun silk fibroin scaffolds. When these results are factored into the useful mechanical properties and slow degradability of this protein biomaterial matrix, potential utility in tissue-engineered blood vessels can be envisioned.
研究了人主动脉内皮细胞（HAEC）和人冠状动脉平滑肌细胞（HCASMC）对电纺丝素蛋白支架的反应，以评估其在血管组织工程中的潜力。细胞增殖研究支持了这种生物材料基质对HAEC和HCASMC的实用性。基于扫描电子显微镜（SEM）和共聚焦显微镜观察，接种后5天内可观察到HCASMC在随机非织造纳米纤维丝支架上的排列和伸长。接种后第4天，HAEC在支架上形成短索状结构，第7天则显示出具有可识别管腔的复杂相互连接的毛细管网络。通过免疫细胞化学和实时逆转录-聚合酶链反应（RT-PCR）分别在蛋白质和转录水平检测细胞特异性标志物（包括HAEC的CD146、VE-钙黏蛋白、血小板内皮细胞黏附分子-1（PECAM-1）和血管性血友病因子（vWF），以及HCASMC的平滑肌肌球蛋白重链2（SM-MHC2）和平滑肌肌动蛋白（SM-肌动蛋白））的存在，证实了丝素蛋白支架上细胞表型的保留。基于蛋白质和转录水平I型胶原表达的定量分析，也证实了HCASMCs细胞外基质（ECM）的形成。结果表明血管细胞与电纺丝素蛋白支架之间存在良好的相互作用。当将这些结果与这种蛋白质生物材料基质有用的机械性能和缓慢的降解性相结合时，可以设想其在组织工程血管中的潜在用途。
