Bovine fetal serum

Human Gingival Fibroblasts—HGFs (HGF; Applied Biological Materials Inc., Richmond, BC, Canada) were cultured in a supplemented Dulbecco’s Modified Eagle’s Medium—DMEM (Lonza^®, Basel, Switzerland) with 10% Bovine Fetal Serum (Biowest^®, Nuaillé, France) and 1% Penicillin with streptomycin (G255 Applied Biological Materials Inc., Richmond, BC, Canada).
Human Fetal Osteoblasts—hFOBs 1.19 (CRL-11372^TM; American Culture Collection, Manassas-ATCC^®, Manassas, VA, USA) were cultured in a mixture (1:1 v/v) of DMEM (Dulbecco’s modified Eagle’s Medium-DMEM) from (BiowhittakerTM, LonzaTM, Basel, Switzerland) and Ham’s F-12 Medium (Sigma-Aldrich^® 51651C, St. Louis, MO, USA), in which was added 0.3 mg·mL⁻¹ of G418 (InvivoGgen, Toulouse, France) and 10% of Bovine Fetal Serum (Biowest^®, Nuaillé, France).
Both cell lines were incubated at 37 °C with 5% CO₂ and 98% humidity. When the cells reached approximately 80% confluence, trypsin-EDTA (Lonza, Veners, Belgium) was added to detach them, they were later centrifuged, and the pellet was resuspended in the respective medium. To perform each cell culture assay 1 × 10⁴ cells/mL cells at a 4th passage were seeded in 48-well plates containing sterile sample (Corning, NY, USA).

Human fetal osteoblasts (hFOB 1.19—CRL-11372^TM; ATCC^®, American Culture Collection, Manassas, VA, USA) were cultured in 75 cm² culture flask (VWR^TM, Radnor, PA, USA) in an atmosphere of 98% humidity and 5% CO₂ at 37 °C with a culture medium constituted of a mixture (1:1 v/v) of Ham’s F12 Medium (Sigma-Aldrich^®, St. Louis, MO, USA) and Dulbecco’s Modified Eagle’s Medium—DMEM (Biowhittaker^TM, Lonza^TM, Basel, Switzerland) supplemented with 10% bovine fetal serum (Biowest, Nuaillé, France) and 0.3 mg/mL G418 (InvivoGen, Toulouse, France).
Immortalized human gingival fibroblasts (hTERT—T0026; Applied Biological Materials Inc., Richmond, BC, Canada) were cultured under the same atmospheric conditions previously described for osteoblasts. The culture medium was constituted of Dulbecco’s Modified Eagle’s Medium (DMEM) (Biowhittaker^TM, Lonza^TM, Switzerland) with 10% of bovine fetal serum (Biowest, Nuaillé, France) and 1% of penicillin-streptomycin (Lonza^TM, Basel, Switzerland).
Cells were trypsinized using trypsin-EDTA (Lonza^TM, Basel, Switzerland) and cells were seeded on discs, distributed in 24-well culture plates (Corning, Corning, NY, USA) at a density of 1 × 10⁴ cells/well. All biological assays were conducted using a fifth passage. For each assay a negative control (cells cultured directly on treated polystyrene surface of the well) was used.