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Primary cilia of tubular cells are sensory organelles. Bending of the primary cilia with shear stress from urinary flow results in the elevation of intracellular calcium levels and activation of signaling pathways that maintain kidney function. Elongation of primary cilia is reported to occur due to oxidative stress, which is a major cause of ischemia-reperfusion injury and is accompanied by decreased kidney function. However, in the context of diminished kidney function, this elongation is yet to be investigated. In this study, we developed a new microfluidic device to monitor changes in the intracellular calcium levels while modulating shear stress on the cilia under different degrees of oxidative stress. The microfluidic device was designed to expose even shear stress in the observed area while supplying drugs in four different stepwise concentrations. The results showed that primary cilia were elongated by hydrogen peroxide, which induces oxidative stress. It was also observed that the elongated primary cilia were more sensitive to shear stress than those with normal morphology. This microfluidic device could, thus, be useful in the analysis of the morphology of the primary cilia, under low perfusion conditions.

Intracellular calcium response of primary cilia of tubular cells to modulated shear stress under oxidative stress