Adhesion of MC3T3-E1 cells to RGD peptides of different flanking residues: Detachment strength and correlation with long-term cellular function
Adhesion of MC3T3-E1 cells to RGD peptides of different flanking residues: Detachment strength and correlation with long-term cellular function
复制标题
DOI：
10.1002/jbm.a.31065
复制
发表时间：
2007-04-01
影响因子：
4.9
通讯作者：
Ducheyne, Paul
中科院分区：
文献类型：
--
作者：
Lee, Mark H.;Adams, Christopher S.;Ducheyne, Paul
我们合成了一系列 RGD 肽，并使用改进的基于马来酰亚胺的缀合物技术将它们固定到胺功能自组装单层上，该技术最大限度地减少了非特异性相互作用。使用转盘装置，发现不同侧翼残基的RGD肽的分离强度(tau(50))的趋势：RGDSPK > RGDSVVYGLR 近似于RGDS > RGES。使用封闭单克隆抗体，细胞对肽的粘附被证明主要是α(v)-整合素介导的。相比之下，表面密度相似的纤连蛋白 (Fn) 包被基质上的细胞 tau(50) 值高出 6-7 倍，并且涉及 α(5)β(1) 和 α(v)β(3) 整合素。与 RGE 和未修饰的底物相比，RGD 肽 1 小时后细胞扩散增强。然而，不同 RGD 肽之间没有观察到显着差异。还通过测量碱性磷酸酶 (ALP) 活性和矿物质沉积来评估 MC3T3-EI 细胞的长期功能。在四种肽中，RGDSPK 在 11 天后表现出最高水平的 ALP 活性，在 15 天后表现出最高的矿化水平，并分别在 15 天和 24 天后达到与 Fn 底物相当的水平。这些发现共同说明了通过 RGD 肽设计增强细胞粘附和功能的优点和局限性。 (c) 2006 Wiley periodicals, Inc. J Biomed Mater Res 81A：150-160，2007。
We synthesized a series of RGD peptides and immobilized them to an amine-functional self-assembled monolayer using a modified maleimide-based conjugate technique that minimizes nonspecific interactions. Using a spinning disc apparatus, a trend in the detachment strength (tau(50)) of RGD peptides of different flanking residues was found: RGDSPK > RGDSVVYGLR approximate to RGDS > RGES. Using blocking monoclonal antibodies, cellular adhesion to the peptides was shown to be primarily alpha(v)-integrin-mediated. In contrast, the tau(50) value of the cells on fibronectin (Fn)coated substrates of similar surface density was 6-7 times higher and involved both alpha(5)beta(1) and alpha(v)beta(3) integrins. Cellular spreading was enhanced on RGD peptides after 1 h when compared to RGE and unmodified substrates. However, no significant differences were observed between the different RGD peptides. Long-term function of MC3T3-EI cells was also evaluated by measuring alkaline phosphatase (ALP) activity and mineral deposition. Among the four peptides, RGDSPK exhibited the highest level of ALP activity after 11 days and mineralization after 15 days and reached comparable levels as Fn substrates after 15 and 24 days, respectively. These findings collectively illustrate both the advantages and limitations of enhancing cellular adhesion and function by the design of RGD peptides. (c) 2006 Wiley Periodicals, Inc. J Biomed Mater Res 81A: 150-160,2007.