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基于具有不同结构和组成特征的表面结合蛋白纳米颗粒的高通量细胞运动性研究

High-Throughput Cell Motility Studies on Surface-Bound Protein Nanoparticles with Diverse Structural and Compositional Characteristics.

作者信息

Tatkiewicz Witold I, Seras-Franzoso Joaquin, García-Fruitós Elena, Vazquez Esther, Kyvik Adriana R, Ventosa Nora, Guasch Judith, Villaverde Antonio, Veciana Jaume, Ratera Imma

机构信息

Department of Molecular Nanoscience and Organic Materials, Institut de Ciència de Materials de Barcelona (ICMAB-CSIC), Campus UAB, 08193 Bellaterra, Spain.

CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), 08193 Bellaterra, Spain.

出版信息

ACS Biomater Sci Eng. 2019 Oct 14;5(10):5470-5480. doi: 10.1021/acsbiomaterials.9b01085. Epub 2019 Sep 23.

Abstract

Eighty areas with different structural and compositional characteristics made of bacterial inclusion bodies formed by the fibroblast growth factor (FGF-IBs) were simultaneously patterned on a glass surface with an evaporation-assisted method that relies on the coffee-drop effect. The resulting surface patterned with these protein nanoparticles enabled to perform a high-throughput study of the motility of NIH-3T3 fibroblasts under different conditions including the gradient steepness, particle concentrations, and area widths of patterned FGF-IBs, using for the data analysis a methodology that includes "heat maps". From this analysis, we observed that gradients of concentrations of surface-bound FGF-IBs stimulate the total cell movement but do not affect the total net distances traveled by cells. Moreover, cells tend to move toward an optimal intermediate FGF-IB concentration (i.e., cells seeded on areas with high IB concentrations moved toward areas with lower concentrations and vice versa, reaching the optimal concentration). Additionally, a higher motility was obtained when cells were deposited on narrow and highly concentrated areas with IBs. FGF-IBs can be therefore used to enhance and guide cell migration, confirming that the decoration of surfaces with such IB-like protein nanoparticles is a promising platform for regenerative medicine and tissue engineering.

摘要

利用依赖咖啡滴效应的蒸发辅助方法,在玻璃表面同时构建了由成纤维细胞生长因子形成的具有不同结构和组成特征的80个细菌包涵体区域(FGF-IBs)。用这些蛋白质纳米颗粒构建的表面,能够在不同条件下(包括梯度陡度、颗粒浓度和图案化FGF-IBs的区域宽度)对NIH-3T3成纤维细胞的运动性进行高通量研究,并使用包括“热图”在内的方法进行数据分析。通过该分析,我们观察到表面结合的FGF-IBs浓度梯度刺激细胞的整体运动,但不影响细胞移动的总净距离。此外,细胞倾向于向最佳的中间FGF-IB浓度移动(即接种在高IB浓度区域的细胞向低浓度区域移动,反之亦然,直至达到最佳浓度)。此外,当细胞接种在狭窄且IB浓度高的区域时,可获得更高的运动性。因此,FGF-IBs可用于增强和引导细胞迁移,证实用此类IB样蛋白质纳米颗粒修饰表面是再生医学和组织工程的一个有前景的平台。

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