Methanol

After 14 days, the formation of mineralization nodules was assessed by alizarin red (Sigma) stain, following the manufacturer's instructions. Briefly, cells were fixed in 4% formaldehyde for 15 min, rinsed in PBS, and stained with 40 mM alizarin red staining reagent for 30 min. Cells were washed four times with distilled water, followed by an immediate 15-min rinse with PBS to reduce non-specific dying. The resulting samples were qualitatively analyzed and photographed under an inverted microscope^{80 (link)}. For quantitative analysis, a 4:1 ratio of 10% acetic acid (Dinâmica) and methanol (Dinâmica) was added to each well to dissolve calcified nodules. Then, the supernatant was read by a spectrophotometer at 405 nm^{82 (link)}. The test was performed in technical triplicate and three individual replicates, conducted on different days.

Antimony(III) potassium tartrate trihydrate from Vetec (Duque de Caxias, RJ, Brazil) and anhydrous quercetin from Sigma-Aldrich (St. Louis, MO, USA) for SbQ complex. For antibacterial assay, it was used brain heart infusion (BHI), rutin hydrate both Sigma-Aldrich Co. (Saint Louis, MO, USA), triphenyltetrazolium chloride (TTC) from Inlab (São Paulo, SP, Brazil) and NaCl from Synth (Diadema, SP, Brazil). Cell lines of bacteria Staphylococcus aureus (CCCD-S007) and Escherichia coli (ATCC-25922). The solvents used were ultrapure water from Merck (Darmstadt, Germany), methanol from Dinâmica (Indaiatuba, SP, Brazil) and dimethylsulphoxide (DMSO) from Synth (Diadema, SP, Brazil).

The materials consisted of culture media, bacterial and fungal strains, commercial antibiotics as well as analytical grade reagents. Potato dextrose agar (PDA) and Mueller–Hinton broth were purchased from Kasvi (Kasvi, São José dos Pinhais, Brazil). Sabouraud broth and yeast extract were supplied by Himedia (Himedia, Thane, India). Microorganisms’ strains were acquired from Cefar (Cefar Diagnóstica, Jardim Taquaral, Brazil). Levofloxacin, terbinafine and chloramphenicol were obtained from EMS (EMS pharma, Hortolândia, Brazil). Dextrose, methanol, ethyl acetate, FeCl₃, dimethyl sulfoxide (DMSO), 2,3,5-triphenyltetrazoic chloride (TTC) and Folin–Ciocalteu solution were purchased from Dinâmica (Dinâmica, Indaiatuba, Brazil). Resazurin, 2,2-diphenyl-1-picrylhydrazyl (DPPH), quercetin, ascorbic acid, Trolox and gallic acid were obtained from Sigma-Aldrich (Sigma-Aldrich, Saint Louis, MO, USA). 2,4,6-Tripyridyl-s-triazine (TPTZ) was supplied by Merck (Merck, Darmstadt, Germany).