Evaluation of cell binding to collagen and gelatin: a study of the effect of 2D and 3D architecture and surface chemistry.
Evaluation of cell binding to collagen and gelatin: a study of the effect of 2D and 3D architecture and surface chemistry.
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DOI：
10.1007/s10856-016-5763-9
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发表时间：
2016-10
期刊：
影响因子：
--
通讯作者：
Cameron RE
中科院分区：
文献类型：
--
作者：
Davidenko N;Schuster CF;Bax DV;Farndale RW;Hamaia S;Best SM;Cameron RE
对细胞附着于胶原材料的研究常常忽略结合机制的细节——无论是整合素介导的还是非特异性的。在这项工作中，我们使用了具有不同尺寸特征（单层、薄膜和多孔支架）的胶原蛋白和明胶基底，以确定成分、交联（使用碳二亚胺）处理和 2D 或 3D 结构对整合素介导的细胞粘附的影响。通过改变受体表达，使用具有胶原结合整合素的细胞（HT1080和C2C12 L3细胞系，表达α2β1和Rugli表达α1β1）和具有明胶结合受体（αvβ3和α5β1）的亲本细胞系C2C12，研究整合素结合位点的性质，以解释不同蛋白质制剂的生物活性。我们已经证明，由于向胶原蛋白中添加明胶并交联而导致化学特性、构象和游离结合基序（GxOGER 和 RGD）可用性的改变，对细胞粘附这些制剂的能力具有深远的影响。碳二亚胺交联消除了二维和三维结构上的整合素依赖性细胞活性，而三维支架结构也导致即使在严格的清洗方案之后，三维样品上仍然存在高水平的非特异性相互作用。这种由交联促进并归因于细胞截留的现象在对三维基质的生物活性进行任何评估时都应予以考虑。传播数据证实了整合素介导的细胞参与对于基于胶原的组合物的进一步细胞活性的重要性。在这项工作中，我们提供了一种简单但有效的方法来解卷积基质的化学和尺寸特征对蛋白质衍生材料的细胞活性的影响，这将有助于针对特定的组织工程应用调整其生物特性。
Studies of cell attachment to collagen-based materials often ignore details of the binding mechanisms—be they integrin-mediated or non-specific. In this work, we have used collagen and gelatin-based substrates with different dimensional characteristics (monolayers, thin films and porous scaffolds) in order to establish the influence of composition, crosslinking (using carbodiimide) treatment and 2D or 3D architecture on integrin-mediated cell adhesion. By varying receptor expression, using cells with collagen-binding integrins (HT1080 and C2C12 L3 cell lines, expressing α2β1, and Rugli expressing α1β1) and a parent cell line C2C12 with gelatin-binding receptors (αvβ3 and α5β1), the nature of integrin binding sites was studied in order to explain the bioactivity of different protein formulations. We have shown that alteration of the chemical identity, conformation and availability of free binding motifs (GxOGER and RGD), resulting from addition of gelatin to collagen and crosslinking, have a profound effect on the ability of cells to adhere to these formulations. Carbodiimide crosslinking ablates integrin-dependent cell activity on both two-dimensional and three-dimensional architectures while the three-dimensional scaffold structure also leads to a high level of non-specific interactions remaining on three-dimensional samples even after a rigorous washing regime. This phenomenon, promoted by crosslinking, and attributed to cell entrapment, should be considered in any assessment of the biological activity of three-dimensional substrates. Spreading data confirm the importance of integrin-mediated cell engagement for further cell activity on collagen-based compositions. In this work, we provide a simple, but effective, means of deconvoluting the effects of chemistry and dimensional characteristics of a substrate, on the cell activity of protein-derived materials, which should assist in tailoring their biological properties for specific tissue engineering applications.
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影响因子：
9.7
作者：
Davidenko N;Schuster CF;Bax DV;Raynal N;Farndale RW;Best SM;Cameron RE
通讯作者：
Cameron RE
影响因子：
14
作者：
Damink, LHHO;Dijkstra, PJ;Feijen, J
通讯作者：
Feijen, J
影响因子：
2.5
作者：
Campbell, Jonathan J.;Botos, Laur-Alexandru;Watson, Christine J.
通讯作者：
Watson, Christine J.
影响因子：
64.5
作者：
Emsley, J;Knight, CG;Liddington, RC
通讯作者：
Liddington, RC
影响因子：
10
作者：
Ashworth JC;Mehr M;Buxton PG;Best SM;Cameron RE
通讯作者：
Cameron RE