Department of Cardiovascular and Thoracic Surgery, Hannover Medical School, Hannover, Germany.
Tissue Eng Part A. 2011 May;17(9-10):1253-61. doi: 10.1089/ten.TEA.2010.0375. Epub 2011 Feb 3.
Tissue-engineered arterial vessels (TEAV) offer substantial advantages in small-calibered human-bypass-grafting and vascularized scaffold applications. However, histological composition of TEAV must allow for functional properties, such as vasomotoricity. Aim of this study was to characterize human TEAVs regarding morphology and vasomotoricity.
Three groups containing segments of porcine carotid artery < 5 mm in diameter (native [NA, n = 6], decellularized [DA, n = 6], and decellularized/reseeded in a bioreactor [RA, n = 7] with human vascular endothelial [hvECs] and smooth muscle cells [hvSMCs]) were examined. Light and scanning electron microscopy were applied, and hvSMCs- and hvECs-associated Vasomotoricity Test conducted in Krebs-solution was used for characterization of revitalized TEAVs.
Morphologic examination showed cell-free extracellular matrix in DAs. Light microscopy demonstrated intact extracellular matrix components in circle-layered formation in cross sections of DAs. RAs showed small cells migrating along the remaining medial fiber structures and flat cell layers at the luminal site, identified as hvECs and hvSMCs with lower CD-31 and α-actin signaling than controls. Scanning electron microscopy showed intact flat cell layers on luminal surfaces of RAs and dense hvSMCs at their media site. DAs showed decreasing strain after stimulation. RAs retrieved vasomotoricity compared to DAs, but showed reduced contraction and incomplete relaxation compared to NAs.
This study shows that revitalization of DA with human vascular cells resembles NA-like morphology and can ensure vasomotoricity of TEAVs.
组织工程化的动脉血管(TEAV)在小口径的人体旁路移植和血管化支架应用中具有显著优势。然而,TEAV 的组织学组成必须允许功能性特性,如血管运动性。本研究的目的是描述人类 TEAV 的形态和血管运动性。
三组含有直径<5 毫米的猪颈动脉段(天然[NA,n=6]、脱细胞[DA,n=6]和在生物反应器中脱细胞/再种植[RA,n=7]与人类血管内皮[hvECs]和平滑肌细胞[hvSMCs])。应用光镜和扫描电镜,并使用在 Krebs 溶液中进行的 hvSMCs 和 hvECs 相关的血管运动性测试来表征再生的 TEAV。
形态学检查显示 DAs 中无细胞的细胞外基质。光镜显示 DAs 横截面上的圆形层状形成中有完整的细胞外基质成分。RAs 显示小细胞沿着残留的中膜纤维结构迁移,并在腔面形成扁平的细胞层,鉴定为 hvECs 和 hvSMCs,其 CD-31 和 α-肌动蛋白信号比对照低。扫描电镜显示 RAs 腔面有完整的扁平细胞层,中膜部位有密集的 hvSMCs。DAs 在刺激后应变减小。RAs 恢复了与 DAs 相比的血管运动性,但与 NA 相比,收缩减少,松弛不完全。
本研究表明,用人类血管细胞再生活体细胞脱细胞基质类似于 NA 样形态,并能确保 TEAV 的血管运动性。