Laminin Peptide-Immobilized Hydrogels Modulate Valve Endothelial Cell Hemostatic Regulation.
Laminin Peptide-Immobilized Hydrogels Modulate Valve Endothelial Cell Hemostatic Regulation.
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DOI：
10.1371/journal.pone.0130749
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Grande-Allen KJ
中科院分区：
文献类型：
--
作者：
Balaoing LR;Post AD;Lin AY;Tseng H;Moake JL;Grande-Allen KJ
瓣膜内皮细胞（VEC）相对于其他类型的血管内皮细胞具有独特的表型反应，并且具有受瓣膜组织变化影响的高度敏感的止血功能。此外，环境因素对VEC止血功能的影响尚未明确。本研究利用聚（乙二醇）二丙烯酸酯（PEGDA）水凝胶平台来评估基质刚度和细胞黏附配体对VEC表型以及止血基因表达的影响。分子量（MW）为3.4、8和20 kDa的水凝胶聚合成不同刚度的平台，巯基修饰的细胞黏附肽与水凝胶表面的丙烯酸酯基团共价结合。肽RKRLQVQLSIRT（RKR）是一种从层粘连蛋白衍生的 syndecan - 1结合配体，层粘连蛋白是一种三聚体蛋白和基底膜基质成分。相反，RGDS是一种在许多细胞外基质（ECM）蛋白（包括纤连蛋白、纤维蛋白原和血管性血友病因子（VWF））中发现的整合素结合肽。VECs黏附在所有涂有RKR的水凝胶 - MW组合上并形成稳定的单层。涂有RGDS的平台支持VEC在RGDS - 3.4 kDa和RGDS - 8 kDa水凝胶上的黏附和生长。在较软的RKR - 8 kDa和RKR - 20 kDa水凝胶平台上培养的VECs，其所有抗血栓（含ADAMTS - 13、组织因子途径抑制剂和组织纤溶酶原激活剂）和血栓（含VWF、组织因子和P - 选择素）蛋白的基因表达明显高于在涂有RGDS的水凝胶和组织培养聚苯乙烯对照上培养的VECs。受刺激的VECs在其各自培养条件下比未受刺激的VECs促进更多的血小板黏附；然而，在RGDS - 3.4 kDa凝胶上受刺激的VECs相对于RKR - 凝胶组对刺激的反应性较低。因此，syndecan结合的、源自层粘连蛋白的肽促进了VEC在较软水凝胶上的稳定黏附，并维持了VEC表型和天然止血功能。总之，利用源自基底膜ECM的非整合素黏附肽序列可能重现平衡的VEC功能，并可能有利于瓣膜植入物的内皮化。
Valve endothelial cells (VEC) have unique phenotypic responses relative to other types of vascular endothelial cells and have highly sensitive hemostatic functions affected by changes in valve tissues. Furthermore, effects of environmental factors on VEC hemostatic function has not been characterized. This work used a poly(ethylene glycol) diacrylate (PEGDA) hydrogel platform to evaluate the effects of substrate stiffness and cell adhesive ligands on VEC phenotype and expression of hemostatic genes. Hydrogels of molecular weights (MWs) 3.4, 8, and 20 kDa were polymerized into platforms of different rigidities and thiol-modified cell adhesive peptides were covalently bound to acrylate groups on the hydrogel surfaces. The peptide RKRLQVQLSIRT (RKR) is a syndecan-1 binding ligand derived from laminin, a trimeric protein and a basement membrane matrix component. Conversely, RGDS is an integrin binding peptide found in many extracellular matrix (ECM) proteins including fibronectin, fibrinogen, and von Willebrand factor (VWF). VECs adhered to and formed a stable monolayer on all RKR-coated hydrogel-MW combinations. RGDS-coated platforms supported VEC adhesion and growth on RGDS-3.4 kDa and RGDS-8 kDa hydrogels. VECs cultured on the softer RKR-8 kDa and RKR-20 kDa hydrogel platforms had significantly higher gene expression for all anti-thrombotic (ADAMTS-13, tissue factor pathway inhibitor, and tissue plasminogen activator) and thrombotic (VWF, tissue factor, and P-selectin) proteins than VECs cultured on RGDS-coated hydrogels and tissue culture polystyrene controls. Stimulated VECs promoted greater platelet adhesion than non-stimulated VECs on their respective culture condition; yet stimulated VECs on RGDS-3.4 kDa gels were not as responsive to stimulation relative to the RKR-gel groups. Thus, the syndecan binding, laminin-derived peptide promoted stable VEC adhesion on the softer hydrogels and maintained VEC phenotype and natural hemostatic function. In conclusion, utilization of non-integrin adhesive peptide sequences derived from basement membrane ECM may recapitulate balanced VEC function and may benefit endothelialization of valve implants.
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影响因子：
6.1
作者：
Fang Ning-tao;Xie Shang-zhe;Pan Luan-feng
通讯作者：
Pan Luan-feng
影响因子：
3.6
作者：
Durbeej, Madeleine
通讯作者：
Durbeej, Madeleine
影响因子：
6.2
作者：
Elbert, DL;Hubbell, JA
通讯作者：
Hubbell, JA
影响因子：
4.6
作者：
Jian, B;Narula, N;Levy, RJ
通讯作者：
Levy, RJ
DOI：
10.1083/jcb.200404171
发表时间：
2004-10-11
期刊：
The Journal of cell biology
影响因子：
--
作者：
Beauvais DM;Burbach BJ;Rapraeger AC
通讯作者：
Rapraeger AC