Mechanotransductive and Hypoxic Effects on Macrophages

To experimentally separate mechanotransductive and hypoxic effects on macrophages that occur concomitantly during OTM, we used lumox cell culture dishes (94.6077.331, Sarstedt), with oxygen-permeable membranes, so that the adherently growing cells could still be supplied with oxygen under experimental pressure application, as pressure and hypoxic conditions are induced concomitantly by the glass disc applied (Figure 1). Approximately 250,000 RAW264.7 macrophages per ml were seeded either onto conventional polystyrene plates (353046, Omnilab) or on lumox plates. After 24 h of preincubation, macrophages were either compressed using glass plates with a defined weight of 2 g/cm² according to an established and published model [7 (link), 16 (link), 17 (link)] or left untreated for further 24 h (Figure 1). To correspond to the results from repeated Experiment 1 with the addition of DMOG, we also repeated this experiment with compressive force treatment for 4 h (Supplemental figures 8-11).