Physical energy-based ultrasound shifts M1 macrophage differentiation towards M2 state

Figure 1 Experimental design for this study in vitro and the method of treating cells. A: At 24 h before lipopolysaccharide (LPS) was added to simulate the inflammatory environment, macrophages at the M1 stage were uniformly subcultured into the six-well plate. Then, The first ultrasound treatment was performed 24 h after the inflammatory environment was maintained. At 48 h, ultrasound was performed on the group requiring a second treatment. The supernatant of culture medium was separated after 24 d of culture (3 d after LPS was added); B: In order to easily adjust the ultrasonic probe to fit the culture holes on the bottom of the six-well plate, we fixed the ultrasonic probe on a sponge pad. Additionally, a box of the same height is used to support the six-hole plate to prevent it from tilting.