Osteoblast-like cell adhesion to bone sialoprotein peptides
Osteoblast-like cell adhesion to bone sialoprotein peptides
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DOI：
10.1016/s0736-0266(03)00180-3
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发表时间：
2004-03-01
影响因子：
2.8
通讯作者：
MacDonald, DE
中科院分区：
文献类型：
--
作者：
Rapuano, BE;Wu, C;MacDonald, DE
多项研究表明，仿生肽可用于新一代假体植入物的设计，以促进植入材料的成功生物整合。在当前的研究中，研究了代表含有 RGD (Arg-Gly-Asp) 的骨唾液蛋白 (BSP) 序列的几种肽对成骨细胞样细胞系 (MC3T3-E1) 的体外生物活性，以深入了解 BSP 与骨细胞相互作用的分子基础。将 BSP 残基 283-288、281-290、278-293 和 278-302 涂覆在 96 孔非组织（未处理）培养板的聚苯乙烯表面上，并将它们的成骨细胞粘附特性与作为阳性对照的完整 BSP 和纤连蛋白进行比较。发现 BSP 肽 278-302 和 278-293 的粘附活性最强，在添加浓度为 1 μM 时，粘附细胞数量增加至对照水平的 350%。由于这两种肽的效力相同，因此表明 RGD 结构域之外的区域 294-302 对于细胞结合不是必需的。相比之下，肽283-288和281-290仅在浓度大于200μM时才有活性。用溶液相 RGD 预处理细胞悬液可抑制 50-70% 的肽刺激的粘附，这表明肽的一部分粘附作用是特异性的且由整合素介导，尽管其他非 RGD 侧翼区域可能也参与粘附机制。重要的是，修饰的BSP肽（其中RGD序列第288位的天冬氨酸残基被谷氨酸残基取代以形成RGE）在浓度LIP至200μM时作为细胞粘附刺激物完全失活。因此，尽管非 RGD 侧翼区域具有潜在作用，但完整的 RGD 三肽对于 BSP 肽的所有粘附活性至关重要。由 Elsevier Ltd. 代表骨科研究协会出版。
A number of studies have suggested that biomimetic peptides can be used in the design of a new generation of prosthetic implants to promote the successful biointegration of the implant materials. In the current study, the in vitro bioactivities of several peptides representing RGD (Arg-Gly-Asp)-containing sequences of bone sialoprotein (BSP) toward an osteoblast-like cell line (MC3T3-E1) were examined to provide insight into the molecular basis of BSP's interaction with bone cells. BSP residues 283-288, 281-290, 278-293 and 278-302 were coated on polystyrene surfaces in 96-well non-tissue (untreated) culture plates, and their osteoblast adhesive properties compared to intact BSP and fibronectin as positive controls. BSP peptides 278-302 and 278-293 were found to be the most potent in their adhesive activity, increasing the number of adherent cells to 350% of control levels at an added concentration of 1 muM. Since these two peptides were equivalent in potency, it is suggested that the region 294-302 beyond the RGD domain is not necessary for cell binding. In comparison, peptides 283-288 and 281-290 were only active at concentrations greater than 200 muM. 50-70% of the peptide-stimulated adhesion was inhibited by the pretreatment of cell suspensions with solution phase RGD, suggesting that a portion of the peptides' adhesive effects was specific and integrin-mediated, although other non-RGD flanking regions were probably also involved in the mechanism of adhesion. Importantly, a modified BSP peptide, in which an aspartic acid residue at position 288 of the RGD sequence was replaced by a glutamic acid residue to form RGE, was completely inactive as a cell adhesion stimulus at concentrations LIP to 200 muM. Thus, despite the potential role of non-RGD flanking regions, an intact RGD tripeptide was essential for all of the adhesive activity of the BSP peptides. Published by Elsevier Ltd. on behalf of Orthopaedic Research Society.