Cck 8 solution

Ten microliter of CCK-8 solution (7Sea Biotech, Shanghai, China) was added into MECs seeded in 96-well plates and incubated at 37°C for 4 h to evaluate MEC viability. Then, the optical density value was measured at 450 nm with a Microplate Reader (Bio Tek, United States).

CCK8 assay was used to detect cell viability. PIG1 cells were seeded into 96-well plates at a density of 8000 per well. After corresponding treatments, cells were incubated with 100 μl fresh medium with 10 μl CCK8 solution (#C008, 7Sea biotech, China) for 2 h at 37 °C, and then the optical density (OD) was measured at 450 nm by Model 680 Microplate Reader (Bio-Rad, USA).

The cytotoxicity of the extracts to HDMECs was measured by CCK-8 assay. A total of 1 × 10⁴ HDMECs were seeded in 96-well microtiter plates, incubated in 200 μL of endothelial cell medium and cultured with the extracts for 1, 2, 3, 5, 7, and 10 days. Cells cultured in endothelial cell medium were used as a control. At each time interval, CCK-8 solution (7seabiotech, China) was added to the culture system and incubated for 3 h at 37°C. Subsequently, the cell viability was evaluated by determining the absorbance at 495 nm using a microplate reader (Model 200 PRO, TECAN, United States). All the experiments were repeated thrice.

To simulate moist and liquid wound environments in vivo at 37°C, the sensor chip was immersed in endothelial cell medium for 3, 5, and 7 days and buffer at pH 4, 7, and 10 for 3 days. At each time point, the sensor chip was directly powered, and 30 sets of temperature data were read. The smart dressing immersed in Dulbecco’s modified Eagle medium (DMEM) at 37°C for 24 h, followed by extract solution collection. The cytotoxicity of the extract to L-929 cells was measured by CCK-8 assay. A total of 1 × 10⁴ L-929 cells were seeded in 96-well microtiter plates, incubated in 200 μL of DMEM and cultured with the extract solution for 1, 3, 5, and 7 days. Cells cultured in DMEM were used as a control. At each time interval, CCK-8 solution (7seabiotech, China) was added to the culture system and incubated for 3 h at 37°C. Subsequently, the cell viability was evaluated by determining the absorbance at 495 nm using a microplate reader (Model 200 PRO, TECAN, United States). All the experiments were repeated three times.

Cell proliferation was evaluated by the CCK-8, EdU staining and colony formation assays. For the CCK-8 assay, cells were inoculated in 96-well plates and maintained in an incubator for a specified period of time. 10 μL of CCK-8 solution (7sea biotech, China) was added to each well 3 h prior to measurement at 450 nm. For the EdU staining assay, cells were labeled with EdU staining solution (Abbkine, USA) at 37 °C for 4 h, fixed in 4% formaldehyde and permeabilized with 0.5% Triton X-100. Then the cells were incubated in Click-iT mixture and re-stained with DAPI solution, and finally detected under a fluorescent microscope. For the colony formation assay, cells were seeded in 12-well plates and maintained in an incubator for 8 to 12 days to form the colonies. After fixation and crystal violet staining, the colonies in each well were photographed and then counted by the Image J software.