J022015 Development of Cell Stretch Device for evaluation of Axonal Strain Injury

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  • J022015 軸索ひずみ損傷評価のための細胞引張装置の開発*([J022-01]傷害防止工学(1))

Abstract

Diffuse axonal injury (DAI), a major component of traumatic brain injury, is associated with rapid deformation of brain tissue resulting in the stretching of neural axons. Focal axonal swelling, which is the morphological hallmarks of DAI pathology, leads to the disconnection of neurons from tissues, resulting in cell death. In present study, we developed a stretch device that subjected cultured neuronal cells in which the directions of neurite elongation were controlled with microfluidic culture technique to uniaxial stretch and examined the effect of strains along neurite on the cell damage. Neurites from PC 12 cells were extended at 0, 45 and 90 degrees to stretch direction using a fabricated poly(dimethylsiloxane) (PDMS) piece having microgrooves in combination with PDMS substrate. Following stretch with a strain of 0.22 and a strain rate of 27 s^<-1>, the morphology of same neurites were observed at 5 min-24 h. As a result, the swelling along neurites oriented at 0 degrees increased immediately following stretch and the increase was sustained until 24 h, although the swelling along neurites oriented at 45 and 90 degrees transiently increased within 1 h following stretch. Many rupture of neurites was observed more in neuritis oriented at 0 and 90 degrees than in neuritis oriented at 45 degrees, however, the retraction and disappearance of neurites did not differ in any orientation conditions. These results suggest that the difference in strain along neurite induces different types and degrees of neurite damage and not strain of cell-cultured substrate but strain loaded to neurite is important to evaluation of neuronal injury.

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