Page 63 - Physico-Chemical Niche Conditions for Bone Cells
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Cell and Nucleus Volume
At different time points, live cells exposed to PFF gradually shrank towards the cell center. Cell volume decreased already significantly after 30 min PFF, and after 60 min PFF the cell volume was decreased from 5786 μm3 to 2774 μm3 (Figure 1A). Static control cells were oval- shaped with some clear lamellipodia around the cell border (Figure 1B). Nuclei of control cells were round, with a surface of rolls-and-swells (Figure 1B). PFF-treated cells contained lamellipodia as well, but generally to a lesser extend (Figure 1B). The nucleus of PFF-treated cells was ellipsoid. One hour PFF reduced cell volume by 0.3-fold (p < 0.0001) compared to control cells (Figure 1B). PFF reduced nucleus volume by 0.2-fold (p < 0.0001).
Figure 2. Effect of 1 h PFF on the spreading of MC3T3-E1 osteoblasts. (A) 2D image of fixed static control cells stained with phalloidin for F-actin (green) and DAPI for the nuclei (blue). (B) 2D image of 1 h PFF-treated cells stained with phalloidin and DAPI. (C) Cell spreading area measurement and analysis. (D) Cell shape/elongation measurement and analysis (cell spreading length vs. cell spreading width). N = 175 (control) and 165 (PFF) cells from 4 separate glass slides. CON, control; PFF, pulsating fluid flow. ** p < 0.01. ns, not significant. Bar = 100 μm.
Chapter 3
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