二軸引張応力を負荷した培養神経細胞の損傷評価

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Diffuse axonal injury (DAI), a major component of traumatic brain injury, has been suggested to result from inertial forces applied on the head. DAI is a manifestation of microstructural cellular trauma and is accompanied by distinct morphological changes. Focal axonal swellings are the morphological hallmarks of DAI pathology and lead to the disconnection of neurons from target tissues resulting in neuronal death. Our goal is the understanding of the quantitative relation between strain acting on the axons and generation of axonal swellings. In present study, we developed an in vitro two dimensional stretch device that reproduced axonal swellings of in vivo DAI, and verified the input-output relation of the device. Then using this device, we exposed PC12 cells, which extend structurally axon-like cylindrical protrusions in culture, to 10% or 20% strains and measured the length of neurites and number of swellings in PC12 cells until 48 hours after the exposure to stretch by microscope observation. As a result, the length of neurites transiently shortened at 5 minutes and 1 hour after exposure to strain compared to those before exposure to strain. On the other hand, swellings were generated at 5minutes after exposure to strain and were the most in number at 1 hour after exposure to strain compared to swellings in normal neurites. Moreover, the number of swellings in neurites exposed to 20% strain was significantly larger than that exposed to 10% strain at 5 minutes after exposure to strain. These results suggest that production of axonal swellings correlate with strain magnitude acting on the axons.

二軸引張応力を負荷した培養神経細胞の損傷評価

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