Dulbecco modified eagle medium (dmem)

Mouse 3T3-L1 fibroblast and RAW264.7 macrophage were kindly provided by Dr. Lin of the National Taiwan University and Dr. Tsai of the National Taiwan Normal University (Taipei, Taiwan), respectively. The original cell lines were purchased from Bioresource Collection and Research Center, Food Industry Research and Development Institute (Hsinchu, Taiwan). Mouse 3T3-L1 and RAW264.7 cells were cultured in Dulbecco's modified Eagle's medium (DMEM, Caisson, Smithfield, UT, USA) containing 10% heat-inactivated calf serum (BS, Gibco, Grand Island, NY, USA) and fetal bovine serum (FBS, Genedirex, Las Vegas, NV, USA), respectively. Murine 4T1 breast cancer cells were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) and maintained in DMEM containing 10% heat-inactivated FBS with 1% penicillin/streptomycin/amphotericin B (Caisson) at 37°C in an incubator containing a humidified atmosphere of 5% CO₂. Aspirin was purchased from Sigma (St. Louis, MO, USA), dissolved in dimethyl sulfoxide (DMSO, Sigma) as a stock solution, and then stored at −20°C. The concentration of DMSO in the vehicle group was equal to the 0.25% DMSO in the highest (5 mM) dose of Aspirin.

The tissue of primary human oral fibroblast (HOF) culture was donated by patients who had signed the informed consent form in the dental clinic of the China Medical University Hospital. The HOF cultures were seeded and cultured in Dulbecco’s modified Eagle medium (DMEM; Caisson Laboratories, North Logan, UT, USA) containing 10% fetal bovine serum (FBS) and 1% antibiotic/antimycotic at 37 °C in an atmosphere of 5% CO₂ in 100 mm culture dishes [28 (link)].

The release of Xu Duan was analyzed after immersing scaffolds in 2 mL of Dulbecco’s Modified Eagle Medium (DMEM, Caisson, North Logan, UT, USA) at 37 °C for different time-points. The count of Xu Duan in medium was measured using the Bio-Rad DC Protein Assay kit. The DMEM without materials was used as the control.