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首页> 外文期刊>Cell structure and function >Second Harmonic Generation Reveals Collagen Fibril Remodeling in Fibroblast-populated Collagen Gels
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Second Harmonic Generation Reveals Collagen Fibril Remodeling in Fibroblast-populated Collagen Gels

机译：第二个谐波世代揭示了成纤维细胞填充胶原蛋白凝胶中的胶原蛋白原纤维重塑

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References(42) Cited-By(4) Remodeling of collagen fibrils is involved in a variety of physiological and pathological processes including development, tissue repair, and metastasis. Fibroblast-populated collagen gel contraction has been employed as a model system to investigate the collagen fibril remodeling within three-dimensional collagen matrices. Research on collagen gel contraction is also important for understanding the mechanism underlying connective tissue repair, and for design considerations for engineered tissues in regenerative medicine. Second harmonic generation (SHG) is a non-linier optical effect by which well-ordered protein assemblies, including collagen fibrils, can be visualized without any labeling, and used for a noninvasive imaging of collagen fibrils in the skin. Here we demonstrate that the remodeling of collagen fibrils in the fibroblast-populated collagen gel can be analyzed by SHG imaging with a multiphoton microscope. Two models of collagen gel contraction (freely versus restrained contraction) were prepared, and orientation of fibroblasts, density, diameter, and distribution of collagen fibrils were examined by multiphoton fluorescent and SHG microscopy. Three-dimensional construction images revealed vertical and horizontal orientation of fibroblasts in freely and restrained gel contraction, respectively. Quantitative analysis indicated that collagen fibrils were accumulated within the gel and assembled into the thicker bundles in freely but not restrained collagen gel contraction. We also found that actomyosin contractility was involved in collagen fibril remodeling. This study elucidates how collagen fibrils are remodeled by fibroblasts in collagen gel contraction, and also proves that SHG microscopy can be used for the investigation of the fibroblast-populated collagen gel.
机译：参考文献（42）被引用（4）胶原纤维的重塑涉及多种生理和病理过程，包括发育，组织修复和转移。成纤维细胞填充的胶原蛋白凝胶收缩已被用作模型系统，以研究三维胶原蛋白矩阵内的胶原蛋白原纤维重塑。胶原凝胶收缩的研究对于理解结缔组织修复的机制以及再生医学中工程组织的设计考虑也很重要。二次谐波（SHG）是一种非线性光学效应，通过这种光学效应，包括胶原原纤维在内的排列良好的蛋白质组件可以在没有任何标记的情况下可视化，并用于皮肤中胶原原纤维的非侵入性成像。在这里，我们证明可以通过多光子显微镜的SHG成像来分析成纤维细胞中胶原凝胶中胶原纤维的重塑。制备了两种胶原蛋白凝胶收缩模型（自由收缩与约束收缩），并通过多光子荧光和SHG显微镜检查了成纤维细胞的取向，密度，直径和胶原蛋白原纤维的分布。三维结构图像显示成纤维细胞在自由和约束的凝胶收缩分别在垂直和水平方向。定量分析表明，胶原蛋白原纤维聚集在凝胶中，并以自由但不限制胶原蛋白凝胶收缩的方式组装成较粗的束。我们还发现，放线菌素的收缩性与胶原纤维的重塑有关。这项研究阐明了成纤维细胞在胶原蛋白凝胶收缩中如何重塑胶原蛋白原纤维，并且还证明了SHG显微镜可以用于研究成纤维细胞填充的胶原蛋白凝胶。

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