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Results 36 3.2.5.

Results 36 3.2.5. Time-laps analysis of cell behaviour Cell migration is an important mechanism during bone turnover and especially wound healing. Therefore, we investigated the influence of oxygen concentration onto migration of hMSCs on polystyrene and collagen I, fibronectin or laminin coated glass slides respectively by time-lapse analysis for 72 h. All data was relatively calculated against Normoxia. HMSCs in 21% oxygen bared a mean velocity of 100.0 ± 26.4% on normal cell culture polystyrene. In contrast, hypoxic observed cells highly significant increased the relative velocity to 164.0 ± 43.2% (p < 0.0001, Fig. 18) on polystyrene. Coating with collagen I ended up with 100.0 ± 26.0% for normoxic cells. The reduction of oxygen concentration to 2% with collagen I coated surfaces showed a slightly decreased, but still highly significant difference of 145.2 ± 46.5% (p < 0.0001) compared to the normoxic control. Interestingly, fibronectin coating came up with a minimal, but significant increased relative migration velocity for observed cells (100.0 ± 26.76% against 112.2 ± 35.8%, p=0.0063). Moreover, hypoxic cells had a significant higher relative velocity 140.7 ± 40.7% (p < 0.0001) compared to the Normoxic control (100.0 ± 27.9%) on laminin coated glass slides. Exemplary polar plots of the Eucledian distance are shown in figure 19. Absolute cell migration velocities are given in supplementary table 3. Fig. 18: Comparison of hMSC migration in normoxic and hypoxic environment on different proteins. Normoxic cells were assigned to 100%. At least 100 cells from two runs were included. Mean & SD, **

Results 37 Fig. 19: Exemplary cell tracks of 10 cells per culture condition and protein coating. Only cells which were observable and did not divide for 24h were included. Polar plots show a non-directed movement for all experiments. The Eucledian distance (distance between start point and end point) was nearly doubled for polystyrene and collagen I or laminin coating in hypoxic conditions. Fibronectin coating

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