The preosteoblast MC3T3-E1 cells (CRL-2593, American Type Culture Collection, Rockville, MD) and the human normal skin fibroblast cells (provided by Z. Qin from the Department of Plastic and Reconstructive Surgery at Peking University Third Hospital) were cultured on the surface of hydrogels at a density of 105 cells/ml for 48 hours. LIVE/DEAD staining solution consisted of calcein-AM (2 μM), and propidium iodide (4 μM) solution was used to incubate hydrogels for 30 min at room temperature. Afterward, these hydrogels were washed three times with phosphate-buffered saline (PBS). Then, hydrogels were observed under a fluorescence microscope (Nikon C2-SIM, Japan). Cell viability was measured by the CCK-8 method. The human normal skin fibroblast cells were cultured in a 48-well plate with or without (control) the hydrogel with an initial density of 5 × 104 cells/ml. At different intervals (1, 4, and 7 days), the culture medium was replaced with medium containing CCK-8 reagent and cells were subsequently incubated at 37°C for 3 hours. The optical density (OD) was measured at 450 nm [cell viability = OD(with hydrogel)/OD(control)].

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