Human gingival fibroblasts hgfs

No further purification was required as all chemicals were obtained from commercial suppliers. Zn (NO₃) ₂^.6H₂O and 2-Methylimidazole were obtained from Sigma (Aldrich, St. Louis, MO, United States). RIZOL was purchased acquired from Life Technologies (Carlsbad, CA, United States). Takara (Shiga, Japan) supply RNA-iso Plus, Prime Script RT reagent kit and TB Green Premix Ex Taq II kit. Kits for the detection of superoxide dismutase (SOD), catalase (CAT) and total antioxidant volume (T-AOC) were purchased from Solarbio (Beijing, China). In addition to human gingival fibroblasts (HGFs, ScienCell, SanDiego, CA, United States), murine macrophages (RAW 264.7 cell line), Escherichia coli (E. coli) ATCC 25922 and Staphylococcus aureus (S. aureus) ATCC 29213 were obtained from American Type Culture Collection (ATCC, Manassas, VA). Rutin hydrate, Calcian-AM/PI Double Staining Kit and all chemical reagents and other biological kits were obtained from Dalian Meilun Biotechnologies Co., Ltd. No additional purification of all reagents used.

SCC4 and SCC25 human OSCC cell lines were kindly provided by Dr. J. F. Liu (School of Oral Hygiene, College of Oral Medicine, Taipei Medical University, Taipei, Taiwan). SCC4 and SCC25 cells were grown in DMEM/F12 supplemented with 10% fetal bovine serum (FBS), 2 mM glutamine and 0.4 μg/ml hydrocortisone. Cells were maintained as monolayer cultures in a humidified atmosphere of 5% CO₂ at 37°C. In addition, human gingival fibroblasts (HGFs) and human oral keratinocytes (HOKs) were purchased from ScienCell Research Laboratories (San Diego, CA). They were cultured in fibroblast medium (ScienCell Research Laboratories, San Diego, CA) and oral keratinocyte medium (ScienCell Research Laboratories, San Diego, CA), respectively, supplemented with 10% FBS, 100 μg/ml streptomycin, and 100 U/ml penicillin at 37°C in 5% CO₂. Experiments were performed with cells from passages 3 to 8. Prior to treatment or stimulation with reagents, cells were serum starved for 24 h.