Heilshorn Sarah C, DiZio Kathleen A, Welsh Eric R, Tirrell David A
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125, USA.
Biomaterials. 2003 Oct;24(23):4245-52. doi: 10.1016/s0142-9612(03)00294-1.
This study examines the spreading and adhesion of human umbilical vein endothelial cells (HUVEC) on artificial extracellular matrix (aECM) proteins containing sequences derived from elastin and fibronectin. Three aECM variants were studied: aECM 1 contains lysine residues periodically spaced within the protein sequence and three repeats of the CS5 domain of fibronectin, aECM 2 contains periodically spaced lysines and three repeats of a scrambled CS5 sequence, and aECM 3 contains lysines at the protein termini and five CS5 repeats. Comparative cell binding and peptide inhibition assays confirm that the tetrapeptide sequence REDV is responsible for HUVEC adhesion to aECM proteins that contain the CS5 domain. Furthermore, more than 60% of adherent HUVEC were retained on aECM 1 after exposure to physiologically relevant shear stresses (</=100dynes/cm(2)). Finally, the levels of thrombogenic markers (tissue plasminogen activator and plasminogen activator inhibitor-1) secreted by HUVEC monolayers on aECM 1 were found to be similar to those secreted by HUVEC monolayers cultured on fibronectin. These characteristics, along with the physical strength and elasticity of crosslinked films prepared from these materials, make aECM proteins promising candidates for application in small-diameter vascular grafts.
本研究考察了人脐静脉内皮细胞（HUVEC）在含有源自弹性蛋白和纤连蛋白序列的人工细胞外基质（aECM）蛋白上的铺展和黏附情况。研究了三种aECM变体：aECM 1在蛋白序列中含有周期性间隔的赖氨酸残基以及纤连蛋白CS5结构域的三个重复序列，aECM 2含有周期性间隔的赖氨酸以及一个混乱的CS5序列的三个重复序列，aECM 3在蛋白末端含有赖氨酸以及五个CS5重复序列。比较细胞结合和肽抑制试验证实，四肽序列REDV负责HUVEC对含有CS5结构域的aECM蛋白的黏附。此外，在暴露于生理相关剪切应力（≤100达因/平方厘米）后，超过60%的黏附HUVEC保留在aECM 1上。最后，发现HUVEC单层在aECM 1上分泌的血栓形成标志物（组织纤溶酶原激活物和纤溶酶原激活物抑制剂-1）水平与在纤连蛋白上培养的HUVEC单层分泌的水平相似。这些特性，连同由这些材料制备的交联膜的物理强度和弹性，使得aECM蛋白成为应用于小直径血管移植物的有前景的候选物。
