HUMAN ENDOTHELIAL-CELL INTERACTIONS WITH SURFACE-COUPLED ADHESION PEPTIDES ON A NONADHESIVE GLASS SUBSTRATE AND 2 POLYMERIC BIOMATERIALS
HUMAN ENDOTHELIAL-CELL INTERACTIONS WITH SURFACE-COUPLED ADHESION PEPTIDES ON A NONADHESIVE GLASS SUBSTRATE AND 2 POLYMERIC BIOMATERIALS
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DOI：
10.1002/jbm.820250209
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发表时间：
1991-02-01
期刊：
影响因子：
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通讯作者：
HUBBELL, JA
中科院分区：
文献类型：
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作者：
MASSIA, SP;HUBBELL, JA
研究了人脐静脉内皮细胞（HUVECs）在与细胞粘附肽Arg-Gly-Asp （RGD）和Tyr-IIe-Gly-Ser-Arg （YIGSR）共价接枝的基质上的附着、扩散、扩散速率、局灶接触形成和细胞骨架组织。这种方法被用来提供即使在没有血清蛋白的情况下也能粘附在细胞上的底物，并且没有事先用细胞粘附分子（CAM）家族的蛋白对表面进行预处理。这种方法被用于显著提高基质的细胞粘附性，否则细胞不粘附，并通过提供稳定结合的粘附配体来改善细胞与细胞粘附材料相互作用的控制。在改性之前，将糖相玻璃作为模型细胞非粘附基板进行了研究，并将聚对苯二甲酸乙二醇酯（PET）和聚四氟乙烯（PTFE）作为生物医学应用的代表性材料进行了研究。利用三酰氯化学将肽的n端胺与表面羟基部分偶联。在肽接枝之前，PET和PTFE表面羟基化得到PETOH和PTFE- oh。PET-OH的细胞黏附性较弱，而PTFE-OH的细胞黏附性比天然聚合物强得多。c端酪氨酸残基的放射碘化用于定量肽偶联到表面的量，这些量在糖相玻璃上为12.1 pmol/cm2，在PETOH上为139 fmol/cm2，在PTFE-OH上为31 fmol/cm2。尽管即使在血清存在的情况下，糖相玻璃也不支持粘附或扩散，但RGD-和yigsr -移植的糖相玻璃确实支持粘附和扩散，即使只包括白蛋白的血清蛋白。虽然PET和PTFE-OH在添加血清的培养基中培养时支持粘附，但只有白蛋白存在时，这两种材料都不支持粘附，这表明细胞粘附是由吸附的CAM蛋白介导的。然而，当这些材料被肽接枝时，即使只有白蛋白存在，也会发生广泛的粘连和扩散。由于肽接枝很容易控制且具有时间稳定性，而蛋白质吸附很难精确控制且具有时间动态性，因此肽接枝可能比其他方法更有利于提高细胞对生物材料的长期粘附。
The attachment, spreading, spreading rate, focal contact formation, and cytoskeletal organization of human umbilical vein endothelial cells (HUVECs) were investigated on substrates that had been covalently grafted with the cell adhesion peptides Arg-Gly-Asp (RGD) and Tyr-IIe-Gly-Ser-Arg (YIGSR). This approach was used to provide substrates that were adhesive to cells even in the absence of serum proteins and with no prior pretreatment of the surface with proteins of the cell adhesion molecule (CAM) family. This approach was used to dramatically enhance the cell-adhesiveness of substrates that were otherwise cell-nonadhesive and to improve control of cellular interactions with cell-adhesive materials by providing stably bound adhesion ligands. Glycophase glass was examined as a model cell-nonadhesive substrate prior to modification, and polyethylene terephthalate (PET) and polytetrafluoroethylene (PTFE) were examined as representative materials for biomedical applications. The peptides were surface-coupled by their N-terminal amine to surface hydroxyl moieties using tresyl chloride chemistry. Prior to peptide grafting, the PET and PTFE were surface hydroxylated to yield PETOH and PTFE-OH. The PET-OH was less cell-adhesive and the PTFE-OH was much more cell-adhesive than the native polymers. Radioiodination of a C-terminal tyrosine residue was used to quantify the amount of peptide coupled to the surface, and these amounts were 12.1 pmol/cm2 on glycophase glass, 139 fmol/cm2 on PETOH, and 31 fmol/cm2 on PTFE-OH. Although the glycophase glass did not support adhesion or spreading even in the presence of serum, the RGD- and YIGSR-grafted glycophase glass did support adhesion and spreading, even when the only serum protein that was included was albumin. Although PET and PTFE-OH supported adhesion when incubated in serum-supplemented medium, neither of these materials supported adhesion with only albumin present, indicating that cell adhesion is mediated by adsorbed CAM proteins. When these materials were peptide-grafted, however, extensive adhesion and spreading did occur even when only albumin was present. Since the peptide grafting is quite easily controlled and is temporally stable, while protein adsorption is quite difficult to precisely control and is temporally dynamic, peptide grafting may be advantageous over other approaches employed to improve long-term cell adhesion to biomaterials.