Matrix stiffness regulates migration of human lung fibroblasts

Physiological Reports
Shuichi AsanoYoshinori Hasegawa

Abstract

In patients with pulmonary diseases such as idiopathic pulmonary fibrosis and severe acute respiratory distress syndrome, progressive pulmonary fibrosis is caused by dysregulated wound healing via activation of fibroblasts after lung inflammation or severe damage. Migration of fibroblasts toward the fibrotic lesions plays an important role in pulmonary fibrosis. Fibrotic tissue in the lung is much stiffer than normal lung tissue. Emerging evidence supports the hypothesis that the stiffness of the matrix is not only a consequence of fibrosis, but also can induce fibroblast activation. Nevertheless, the effects of substrate rigidity on migration of lung fibroblasts have not been fully elucidated. We evaluated the effects of substrate stiffness on the morphology, α-smooth muscle actin (α-SMA) expression, and cell migration of primary human lung fibroblasts by using polyacrylamide hydrogels with stiffnesses ranging from 1 to 50 kPa. Cell motility was assessed by platelet-derived growth factor (PDGF)-induced chemotaxis and random walk migration assays. As the stiffness of substrates increased, fibroblasts became spindle-shaped and spread. Expression of α-SMA proteins was higher on the stiffer substrates (25 kPa gel and plastic dishe...Continue Reading

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Methods Mentioned

BETA
Assay
protein assay
transfection
SMA
confocal microscopy

Software Mentioned

NIH ImageJ
MetaMorph
ImageJ
Quantity One

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