Analysis of fibroblast migration dynamics in idiopathic pulmonary fibrosis using image-based scaffolds of the lung extracellular matrix.
Analysis of fibroblast migration dynamics in idiopathic pulmonary fibrosis using image-based scaffolds of the lung extracellular matrix.
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使用基于图像的肺细胞外基质支架分析特发性肺纤维化中的成纤维细胞迁移动力学。
DOI：
10.1152/ajplung.00087.2019
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Campagnola,PaulJ
中科院分区：
文献类型：
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作者：
Tisler,Marisa;Alkmin,Samuel;Chang,Hsin-Yu;Leet,Jon;Bernau,Ksenija;Sandbo,Nathan;Campagnola,PaulJ
特发性肺纤维化（IPF）的特征在于细胞外基质（ECM）中胶原蛋白的深刻重塑，其中纤维变得更致密且更高度排列。然而，尚不清楚胶原基质的这种重新配置如何影响疾病进展。在这里，我们研究的作用，具体的改变，在胶原纤维组织细胞迁移动力学通过使用仿生图像为基础的胶原支架代表正常和纤维化肺，其中的设计是直接从高分辨率的二次谐波发生显微镜图像。支架通过多光子激发（MPE）聚合制造，其中该过程类似于三维打印，除了它以更高的分辨率（约0.5微米）和胶原蛋白和胶原蛋白类似物进行。这些支架接种有早期传代的原代人正常和IPF成纤维细胞，以使细胞内在特征（正常vs. IPF）与ECM结构（正常vs. IPF）对迁移动力学的影响解耦。我们发现，相对于正常组织，高度排列的IPF胶原结构促进了增强的细胞伸长和F-肌动蛋白排列沿着增加的细胞迁移速度和直线性。总的来说，这些数据与调整后的指规数矩阵上的强化联系指导机制一致。虽然观察到细胞内在效应，但对齐的胶原基质形态对这些指标有更大的影响。重要的是，这些肺的仿生模型不能通过传统的制造方法合成。我们认为，MPE基于图像的制造方法将使额外的假设为基础的测试研究的细胞-基质相互作用的背景下，组织纤维化。
Idiopathic pulmonary fibrosis (IPF) is characterized by a profound remodeling of the collagen in the extracellular matrix (ECM), where the fibers become both denser and more highly aligned. However, it is unknown how this reconfiguration of the collagen matrix affects disease progression. Here, we investigate the role of specific alterations in collagen fiber organization on cell migration dynamics by using biomimetic image-based collagen scaffolds representing normal and fibrotic lung, where the designs are derived directly from high-resolution second harmonic generation microscopy images. The scaffolds are fabricated by multiphoton-excited (MPE) polymerization, where the process is akin to three-dimensional printing, except that it is performed at much greater resolution (∼0.5 microns) and with collagen and collagen analogs. These scaffolds were seeded with early passaged primary human normal and IPF fibroblasts to enable the decoupling of the effect of cell-intrinsic characteristics (normal vs. IPF) versus ECM structure (normal vs. IPF) on migration dynamics. We found that the highly aligned IPF collagen structure promoted enhanced cell elongation and F-actin alignment along with increased cell migration speed and straightness relative to the normal tissues. Collectively, the data are consistent with an enhanced contact guidance mechanism on the aligned IPF matrix. Although cell intrinsic effects were observed, the aligned collagen matrix morphology had a larger effect on these metrics. Importantly, these biomimetic models of the lung cannot be synthesized by conventional fabrication methods. We suggest that the MPE image-based fabrication method will enable additional hypothesis-based testing studies of cell-matrix interactions in the context of tissue fibrosis.