Second Harmonic Generation Reveals Collagen Fibril Remodeling in Fibroblast-populated Collagen Gels

Cell Structure and Function

Shigeki Higashiyama²  Yuji Shirakata⁵  Fujio Toki⁴  Naoki Honkura³  Daisuke Nanba⁴  Takeshi Imamura¹ 

[1] Division of Bio-imaging, Proteo-Science Center (PROS), Ehime University;Division of Cell Growth and Tumor Regulation, Proteo-Science Center (PROS), Ehime University;Department of Molecular Medicine for Pathogenesis, Graduate School of Medicine, Ehime University;Senior Research Fellow Center, Ehime University;Department of Dermatology, Graduate School of Medicine, Ehime University

关键词: collagen;    collagen gel contraction;    fibroblasts;    second harmonic generation;    mutiphoton microscopy;   

DOI  :  10.1247/csf.13017

学科分类：分子生物学，细胞生物学和基因

来源: Japan Society for Cell Biology

【 摘 要 】

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.

【 授权许可】

Unknown