Mg 63 cell line

Human osteoblast-like cells MG63 cell line was purchased from ATCC company. The cells were culture in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin at 37°C with 5% CO₂ and passage number 10-15 was used. Bone marrow stromal cells (BMSCs) were extracted from the femoral marrow of 3-week female Sprague–Dawley (SD) rats. The cells were cultured in α-MEM with 10% FBS and 1% of penicillin/streptomycin. BMSCs from passage 3 were used for further experiments.

Osteoblasts (MG-63 cell line; American Type Culture Collection, Rockville, MA) were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat-inactivated FBS (Equitech-Bio Inc., TX, USA), 2 mM glutamine and 100 units/ml penicillin/streptomycin (Invitrogen, Carlsbad, CA) at 37°C in a humidified atmosphere of 5% CO₂ in air.

Dental pulp stem cells (DPSCs) were from our lab. The MG-63 cell line was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). After thawing, these cells were maintained in Minimal Essential Medium α (Gibco, Billings, MT, USA) supplemented with 10% fetal bovine serum (Gibco, Billings, MT, USA), 100 μg/mL streptomycin, and 100 μg/mL penicillin (Sigma-Aldrich, St. Louis, MO, USA) in a 5% CO₂ incubator at 37 °C. A total of 100 mM CUR (Sigma-Aldrich, St. Louis, MO, USA) in dimethyl sulfoxide (DMSO, Sigma-Aldrich, St. Louis, MO, USA) was prepared as a store solution and stored in the dark in a −20 °C fridge. Finally, 5, 10, and 20 μM CUR and 5 mM N-Acetyl-L-cysteine (NAC, Sigma Aldrich, St. Louis, MO, USA) with relative concentrations of CUR were administrated to treat DPSCs and MG-63 cells.

Human osteosarcoma MG-63 cell line was purchased from the American Type culture Collection (ATCC, Manassas, VA, USA). The cells were incubated under standard conditions (37 °C, 5% CO₂, and 95% humidity). MG-63 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Thermo Fisher Scientific, Grand Island, NY, USA) containing 10% heat-inactivated (56 °C and 30 min) fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, Missouri, USA) and 1% penicillin/streptomycin antibiotics (Thermo Fisher Scientific, Grand Island, NY, USA).

Human MSCs were isolated and characterized as previously described (Haddouti et al., 2020a (link); Haddouti et al., 2020b (link); Walter et al., 2020 (link)). In brief, MSCs were harvested from the femur head after hip replacement. MSCs were isolated through gradient centrifugation (800 × g for 30 min without brake) using Biocoll separating solution (Biochrom AG, Berlin, Germany). MSCs were plated in cell culture flasks (Greiner Bio-One GmbH, Frickenhausen, Germany) with Dulbecco’s modified Eagle’s medium (DMEM) (Gibco by Life Technologies, Darmstadt, Germany) containing 10% serum (Bio&SELL GmbH, Feucht/Nürnberg, Germany), 1% L-glutamine, and 1% penicillin–streptomycin (Biochrom AG). Incubation took place under standard conditions at 37°C in a humidified atmosphere with 5% CO₂. The culture medium was changed 2–3 times a week. After confluency, MSCs were passaged and stored at −150°C until needed. The MG63 cell line was purchased from the American Type Culture Collection (ATCC) and cultured under the same conditions as MSCs according to the manufacturer’s instructions. The studies involving human participants were reviewed and approved (project IDs: 122/09 and 102/19) and were conducted in accordance with the approved guidelines as well as the Declaration of Helsinki.