Peptide microarrays for the discovery of bioactive surfaces that guide cellular processes: a single step azide-alkyne "click" chemistry approach.
Peptide microarrays for the discovery of bioactive surfaces that guide cellular processes: a single step azide-alkyne "click" chemistry approach.
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DOI：
10.1039/c4tb00375f
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发表时间：
2014-06
期刊：
影响因子：
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通讯作者：
Douglas Zhang;K. Kilian
中科院分区：
文献类型：
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作者：
Douglas Zhang;K. Kilian
细胞在体内的行为是由一个复杂的微环境，含有许多不同的分子，包括细胞外基质（ECM）蛋白，生长因子和蛋白聚糖指导。控制细胞-材料界面上这些不同组分之间的相互作用对于开发用于生物医学设备和组织工程应用的新材料将是非常宝贵的。我们报告了一个单一的步骤的方法，形成混合肽共轭自组装单分子膜的金使用铜催化的叠氮化物-炔环加成化学研究不同的肽配体对细胞过程的组合效应。我们合成了ECM粘附肽（YIGSR，GRGDS），骨形态发生蛋白7（BMP-7）衍生肽（KPSSAPTQLN）和肝素结合肽（KRSR），并将它们单独和组合排列在金涂层盖玻片上。通过X射线光电子能谱（XPS）和基质辅助激光解吸/电离（MALDI）质谱对SAMs进行了表征，发现阵列肽组合与脂肪源干细胞（ADSC）和小鼠胚胎成纤维细胞（MEF）有差异地结合。我们进一步研究了标准培养和成骨分化培养基中ADSC在含有粘附肽和BMP-7肽组合的SAM上的成骨作用。我们证明了当ADSC单独粘附于BMP-7衍生肽或与ECM粘附肽组合时，成骨标志物Runx 2和骨桥蛋白的表达增强。这里提出的平台能够使用市售的微阵列点样器在单个步骤中固定多个肽，这将证明在制造用于细胞生物学研究和生化测定的生物分子界面中是有用的。
Cell behavior in vivo is guided by a complex microenvironment containing many different molecules including extracellular matrix (ECM) proteins, growth factors, and proteoglycans. Controlling the interaction between these various components at the cell-material interface will be invaluable in developing new materials for biomedical devices and tissue engineering applications. We report a single step approach to forming mixed peptide conjugated self-assembled monolayers on gold using copper-catalyzed azide-alkyne cycloaddition chemistry to study the combinatorial effects of different peptide ligands on cellular processes. We synthesized ECM adhesion peptides (YIGSR, GRGDS), a bone morphogenetic protein 7 (BMP-7) derived peptide (KPSSAPTQLN), and a heparin binding peptide (KRSR), and arrayed them, alone and in combination, onto gold coated coverslips. SAMs were characterized by X-ray photoelectron spectroscopy (XPS) and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry, and arrayed peptide combinations were seen to differentially bind to adipose derived stem cells (ADSCs) and mouse embryonic fibroblasts (MEFs). We further investigated the osteogenesis of ADSCs on SAMs containing combinations of adhesion peptide and BMP-7 peptide in both standard culture and osteogenic differentiation media. We demonstrate enhanced expression of osteogenic markers Runx2 and osteopontin when ADSCs are adherent to BMP-7 derived peptide alone or in combination with ECM adhesion peptides. The platform presented here enables immobilization of multiple peptides in a single step using a commercially available microarray spotter which will prove useful in fabricating biomolecule interfaces for cell biology studies and biochemical assays.