title: Cytotoxicity of Agaricus sylvaticus in non-tumor cells (NIH/3T3) and tumor (OSCC-3) using tetrazolium (MTT) assay.
abstract: The purpose of this study was to assess the cytotoxic effect of the non-fractionated aqueous extract of A. sylvaticus mushroom in cultures of non-tumor cells (NIH3T3) and tumor cells (OSCC-3). The cells were maintained in DMEN cell culture medium added of 10% of fetal bovine serum and 1% antibiotic. For the cytotoxicity test we prepared the aqueous mushroom extract at concentrations of 0.01 mg.ml⁻¹, 0.02 mg.ml⁻¹, 0.04 mg.ml⁻¹, 0.08 mg.ml⁻¹, 0.16 mg.ml⁻¹, and 0.32 mg.ml⁻¹. For the culture, 2 x 10⁵ cells/ml was deposited in 96-well microplates during 24 hour incubation with subsequent exchange of medium by another containing the mushroom concentrations. After 24 hour incubation the medium was discarded and 100 ml of tetrazolium blue (MTT) was added at a concentration of 5 mg.ml⁻¹. The microplates were incubated for 2 h at 37° C. Spectrophotometric analysis was performed using 570 nm wavelength. From the values of the optical densities we determined the drug concentration capable of reducing cell viability by 50%. Therefore, the mushroom A. sylvaticus, at all concentrations tested, did not show cytotoxic effects, once the inhibitory concentration (IC₅₀) obtained for tumor cells OSCC-3 was 0.06194 mg.ml⁻¹, and the IC₅₀ checked for non-tumor cells NIH3T3 was 0,06468 mg.ml⁻¹. This test made it possible to determine that A. sylvaticus mushroom has no cytotoxic effects, suggesting its use safe for human consumption.
authors.affiliation: Instituto Federal Goiano-Campus Urutaí, Brazil. joicevinhal@gmail.com
authors.forename: Joice Vinhal Costa
authors.initials: JV
authors.lastname: Orsine
authors.forename: Luíssa
authors.initials: L
authors.lastname: Marques Brito
authors.forename: Renata Carvalho
authors.initials: RC
authors.lastname: Silva
authors.forename: Maria de Fátima Menezes
authors.initials: Mde F
authors.lastname: Santos Almeida
authors.forename: Maria Rita Carvalho Garbi
authors.initials: MR
authors.lastname: Novaes
mesh_terms: D016475:3T3 Cells; D000364:Agaricus / Q000737:chemistry*; D000818:Animals; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects*; D004396:Coloring Agents; D003470:Culture Media; D004354:Drug Screening Assays, Antitumor; D006801:Humans; D051379:Mice; D009145:Mushroom Poisoning; D013056:Spectrophotometry, Ultraviolet; D013778:Tetrazolium Salts; D013844:Thiazoles
chemical_list: D004396:Coloring Agents; D003470:Culture Media; D013778:Tetrazolium Salts; D013844:Thiazoles; C022616:thiazolyl blue