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Solid tumors, such as osteosarcoma, generally suffer from an inadequate oxygen supply due to the rapid proliferation of cancer cells, incomplete vascularization, and uneven distribution

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When the cells reached approximately 80% confluency, cells were harvested with trypsin and seeded at a density of 2.9 × 10 5 cells per well in 2.5 ml of culture medium into 6 well flat-bottomed plates (Costar, Corning, USA) and incubated for 24 hr