RGD and BMP-2 mimetic peptide crosstalk enhances osteogenic commitment of human bone marrow stem cells
RGD and BMP-2 mimetic peptide crosstalk enhances osteogenic commitment of human bone marrow stem cells
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DOI：
10.1016/j.actbio.2016.03.032
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发表时间：
2016-05-01
影响因子：
9.7
通讯作者：
Laroche, G.
中科院分区：
文献类型：
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作者：
Bilem, I.;Chevallier, P.;Laroche, G.
人骨髓间充质干细胞(HBMSCs)的定位和分化是由其细胞外基质(ECM)中的生物活性分子决定的。一种常见的模拟生理环境的方法是用ECM衍生的多肽来功能化生物材料表面，这些多肽能够招募干细胞并触发它们的线条特异性分化。本研究的目的是在不依赖分化介质的情况下，研究RGD和BMP-2配体的串扰和密度对hBMSCs成骨能力的影响。RGD多肽通过细胞跨膜整合素受体促进细胞黏附，而BMP-2多肽可诱导hBMSCs向成骨细胞分化，BMP-2多肽对应于骨形态发生蛋白-2的第73-92位氨基酸残基。用X射线光电子能谱(XPS)确定了多肽在胺化玻璃上的固定化情况，用荧光显微镜对接枝肽的密度进行了定量，用原子力显微镜(AFM)对表面粗糙度进行了评价。以Stro-1为特异性干细胞标记物，RUNX-2为早期成骨标记物，用免疫组织化学方法检测培养的hBMSCs在RGD和/或BMP-2表面的成骨能力。生物学结果表明，与含有BMP-2的表面相比，双功能化表面增强了hBMSCs的成骨能力，而在RGD表面，细胞主要保持了其僵硬的特性。这些结果表明，RGD和BMP-2模拟肽在不添加成骨因子的情况下具有协同促进hBMSCs成骨的作用。这些发现有助于仿生材料的开发，使人们能够更深入地了解控制干细胞向成骨细胞转化的信号通路。意义的陈述长期以来，科学家们认为间充质干细胞(MSCs)向骨细胞的分化是由生长因子决定的。这篇手稿揭示了在调节MSCs命运方面发挥关键作用的其他配体。具体地说，BMP-2多肽的诱导成骨作用在粘附性RGD多肽的存在下提高了2倍。与以前强调RGD和BMP-2多肽之间的协同合作的工作相比，这项工作的主要优势在于使用原始的人类细胞(HMSCs)和定义明确的仿生材料表面(受控的表面粗糙度和多肽密度)。这项工作为开发定制的体外细胞培养模型提供了有价值的见解，能够针对所需的细胞反应。(C)2016 Acta Materialia Inc.由爱思唯尔有限公司出版。版权所有。
Human bone marrow mesenchymal stem cells (hBMSCs) commitment and differentiation are dictated by bioactive molecules sequestered within their Extra Cellular Matrix (ECM). One common approach to mimic the physiological environment is to functionalize biomaterial surfaces with ECM-derived peptides able to recruit stem cells and trigger their linage-specific differentiation. The objective of this work was to investigate the effect of RGD and BMP-2 ligands crosstalk and density on the extent of hBMSCs osteogenic commitment, without recourse to differentiation medium. RGD peptide promotes cell adhesion via cell transmembrane integrin receptors, while BMP-2 peptide, corresponding to residues 73-92 of Bone Morphogenetic Protein-2, was shown to induce hBMSCs osteoblast differentiation. The immobilization of peptides on aminated glass was ascertained by X-ray Photoelectron Spectroscopy (XPS), the density of grafted peptides was quantified by fluorescence microscopy and the surface roughness was evaluated using Atomic Force Microscopy (AFM). The osteogenic commitment of hBMSCs cultured on RGD and/or BMP-2 surfaces was characterized by immunohistochemistry using STRO-1 as specific stem cells marker and Runx-2 as an earlier osteogenic marker. Biological results showed that the osteogenic commitment of hBMSCs was enhanced on bifunctionalized surfaces as compared to surfaces containing BMP-2, while on RGD surfaces cells mainly preserved their sternness character. These results demonstrated that RGD and BMP-2 mimetic peptides act synergistically to enhance hBMSCs osteogenesis without supplementing the media with osteogenic factors. These findings contribute to the development of biomimetic materials, allowing a deeper understanding of signaling pathways that govern the transition of stem cells towards the osteoblastic lineage.Statement of SignificanceFor a long time, scientists thought that the differentiation of Mesenchymal Stem Cells (MSCs) into bone cells was dictated by growth factors. This manuscript shed light on other ligands that play a crucial role in regulating MSCs fate. In concrete terms, it was demonstrated that the osteoinductive effect of BMP-2 peptide is 2 folds improved in the presence of adhesive RGD peptide. Compared to previous works highlighting this synergistic cooperation between RGD and BMP-2 peptides, the main strength of this work lies to the use of primitive human cells (hMSCs) and well-defined biomimetic material surfaces (controlled surface roughness and peptide densities). This work provides valuable insights to develop custom-designed in vitro cell culture models, capable of targeting the desired cell response. (C) 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.