Sfm 101

The GE1 (RIKEN Cell Bank, Tsukuba, Japan) mouse-derived gingival epithelial cell line was used (Figure 1a). GE1 cells were cultured in basal serum-free medium (SFM-101, Nissui, Tokyo, Japan) containing 1% fetal bovine serum (Biowest, Nuaillé, France) and 10 ng/mL mouse epidermal growth factor (Corning, New York, NY, USA) in a humidified atmosphere with 5% CO₂ at 33 °C. Cells were seeded within a 1 mL volume onto each Ti plate, at a density of 5 × 10⁴ cells per well in a 24 well culture plate (Multiwell 24 well, Corning).

To elucidate the osteoblastic or epithelial cell attachment characteristics toward the treated titanium plates, a cell culture experiment was performed.
The GE1 mouse gingival epithelial cell line was provided by RIKEN BRC (Tsukuba, Japan) through the National Bio-Resource Project of the Ministry of Education, Culture, Sports, Science, and Technology, Japan. GE1 cells were cultivated in SFM-101 (Nissui Pharmaceutical, Tokyo, Japan) containing 1% fetal bovine serum (FBS) supplemented with 10 ng/mL mouse epidermal growth factor at 33 °C in a humidified atmosphere of 5% CO₂ in air for 24 h [20 (link)]. The titanium plates (diameter 15 mm, thickness 1 mm) were pre-treated with 3% FBS for 8 h, and cells were seeded in a 0.5 mL volume onto each titanium plate at a density of 0.5 × 10⁵ cells per titanium plate in a 24-well plate (Falcon Labware, Oxford, UK). The MC3T3-E1 osteoblastic cells (RIKEN BRC) were cultured in alpha-minimum essential medium (Gibco, Grand Island, NY, USA) containing 10% FBS at 37 °C in a humidified atmosphere of 5% CO₂ in air for 1 h.