Morphological changes in osteoblastic cells (MC3T3-E1) due to fluid shear stress: cellular damage by prolonged application of fluid shear stress.

We investigated the effect of fluid shear stress on both changes in morphology and in resting level of cytosolic concentrations of Ca2+ ([Ca2+]i) in MC3T3-E1 cells, osteoblast-like cells derived from the neonatal mouse calvaria. The cells were subjected to fluid shear stress at about 1.0 Pa. Morphological characteristics were compared between stressed and control cells by scanning electron microscopic examination, actin filament evaluation using phalloidin fluorescence and [Ca2+]i measurement using fura-2/AM. The stressed cells changed from a polygonal to a spindle shape after 1-hour exposure to fluid shear stress, whereas control cells remained polygonal, and the stressed cells exhibited an increase in the number of microvilli. The diameter in the long axis of the stressed cells was significantly larger than that of controls, and about half of the stressed cells were oriented nearly along the direction of the fluid shear stress. The arrangement of the actin filaments of the stressed cells changed to parallel, and was positioned along the long axis of the cell process. The resting level of [Ca2+]i in the stressed cells showed a peak at 1 hour after the application of the stress. A more prolonged stress period over 1 hour caused cell shrinkage, an irregularity on the cell surface, and a gradual decrease in the resting level of [Ca2+]i, suggesting that prolonged stress time may cause cellular damage.