Controlled spatial and conformational display of immobilised bone morphogenetic protein-2 and osteopontin signalling motifs regulates osteoblast adhesion and differentiation in vitro.
Controlled spatial and conformational display of immobilised bone morphogenetic protein-2 and osteopontin signalling motifs regulates osteoblast adhesion and differentiation in vitro.
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DOI：
10.1186/1741-7007-8-57
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发表时间：
2010-05-10
期刊：
影响因子：
5.4
通讯作者：
Birch MA
中科院分区：
文献类型：
--
作者：
Mitchell EA;Chaffey BT;McCaskie AW;Lakey JH;Birch MA
调节哺乳动物细胞生长和分化的界面分子机制对生物技术（细胞和细胞产品的生产）和医学（组织工程、假体植入、癌症和发育生物学）具有重要意义。我们在这里证明，使用设计用于在金表面上自组装的可溶性蛋白支架，可以以高度受控的方式在体外将工程蛋白基序稳健地展示给哺乳动物细胞。一种蛋白质经过工程改造，含有一个 C 端半胱氨酸，可以化学吸附金，然后是 12 个氨基酸，形成水溶性线圈，在烷硫醇存在的情况下可以转换为疏水性螺旋。来自骨形态发生蛋白 2 或骨桥蛋白的生物活性基序被添加到该支架蛋白中，并在组装到金表面上时评估其影响细胞功能的能力。数据表明，成骨细胞粘附和对骨形态发生蛋白-2 的短期反应取决于骨桥蛋白衍生的细胞粘附基序的表面密度。此外，来自骨形态发生蛋白的固定细胞相互作用基序在体外支持骨形成超过 28 天（在完全没有其他成骨补充剂的情况下）。此外，使用软光刻方法对该配体进行二维图案化导致了成骨的空间控制。这些数据描述了一种方法，可以在分子水平上剖析固定蛋白配体对细胞行为的影响。这种方法提供了一个耐用的表面，可以评估对细胞活动的短期（数小时或数天）和长期（数周）影响。这种广泛适用的方法可以提供固定配体在细胞活性控制中的作用的机制见解。
The interfacial molecular mechanisms that regulate mammalian cell growth and differentiation have important implications for biotechnology (production of cells and cell products) and medicine (tissue engineering, prosthetic implants, cancer and developmental biology). We demonstrate here that engineered protein motifs can be robustly displayed to mammalian cells in vitro in a highly controlled manner using a soluble protein scaffold designed to self assemble on a gold surface. A protein was engineered to contain a C-terminal cysteine that would allow chemisorption to gold, followed by 12 amino acids that form a water soluble coil that could switch to a hydrophobic helix in the presence of alkane thiols. Bioactive motifs from either bone morphogenetic protein-2 or osteopontin were added to this scaffold protein and when assembled on a gold surface assessed for their ability to influence cell function. Data demonstrate that osteoblast adhesion and short-term responsiveness to bone morphogenetic protein-2 is dependent on the surface density of a cell adhesive motif derived from osteopontin. Furthermore an immobilised cell interaction motif from bone morphogenetic protein supported bone formation in vitro over 28 days (in the complete absence of other osteogenic supplements). In addition, two-dimensional patterning of this ligand using a soft lithography approach resulted in the spatial control of osteogenesis. These data describe an approach that allows the influence of immobilised protein ligands on cell behaviour to be dissected at the molecular level. This approach presents a durable surface that allows both short (hours or days) and long term (weeks) effects on cell activity to be assessed. This widely applicable approach can provide mechanistic insight into the contribution of immobilised ligands in the control of cell activity.
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影响因子：
2.9
作者：
Arnold, M;Cavalcanti-Adam, EA;Spatz, JP
通讯作者：
Spatz, JP
影响因子：
4
作者：
Nohe, A;Keating, E;Petersen, NO
通讯作者：
Petersen, NO
影响因子：
5.3
作者：
Hartung, Anke;Bitton-Worms, Keren;Knaus, Petra
通讯作者：
Knaus, Petra
影响因子：
6.2
作者：
Irvine, DJ;Ruzette, AVG;Griffith, LG
通讯作者：
Griffith, LG
影响因子：
6.2
作者：
Jikko, A;Harris, SE;Damsky, CH
通讯作者：
Damsky, CH