The oral cytotoxicity, antimicrobial and anti-demineralizing effects of a tincture from Bauhinia forficata Link tincture (BFLT) were evaluated in vitro and ex vivo. Susceptibility tests (minimum inhibitory and microbicidal concentrations-MIC and time-kill assay-MMC) were performed against planktonic oral microorganisms. The contents of phenolic compounds were investigated. Cytotoxic potential was evaluated on oral fibroblasts after 1-5 min exposure to BFLT. Blocks of sound bovine enamel (N = 60) were inoculated with a saliva pool and sustained in a multiple plaque growth system for 48 h to form a biofilm. Biofilm blocks were randomly divided into groups-G (n = 10): G1-Baseline (48 h maturation biofilm), G2-BFLT 23.2 mg/mL, G3-Ethanol 81.20 g/mL, G4-Chlorhexidine 0.12%, G5-Growth control, and G6-Blank control. Treatments (50 μL/1 min) were performed once a day for a week. Streptococcus spp. (S) and total microorganism (TM) counts were expressed as Log10 CFU/mL. Biofilm height was evaluated by confocal microscopy analyses (CMA). Final surface hardness was assessed and percentage of microhardness loss (% MHL) was calculated. Results were significant when P < .05. BFLT inhibited all tested microorganisms (MIC = 1.3-23.2 mg/mL) and promoted optical reduction (0.05-0.22 nm) of all microorganisms after 48-h treatment compared with controls. After 5-min treatment, BFLT showed low values of cell death (3.20%). G2-BFLT reduced S (6.61 ± 0.20) and TM (7.14 ± 0.38) compared with G1-Baseline (S = 7.82 ± 0.28; TM = 8.81 ± 0.67) and G5-Growth control (S = 7.48 ± 0.39; TM = 7.89 ± 0.68); but G4-chlororexidine (S = 6.11 ± 0.48; TM = 6.45 ± 0.16) showed the highest antibiofilm activity. CMA was not different among treatment groups. G2 showed lower % MHL compared with G5, although G4 presented the lowest. Results suggest BFLT is beneficial against dental caries, showing antimicrobial effects against a mature dental biofilm and no cytotoxicity.