Objective. To determine the influence of two different fluoride compounds and an in vitro pellicle on KOH-soluble fluoride formation - its retention and resistance to toothbrushing. Material and methods. Forty bovine incisors were randomly assigned to four groups (A-D). Of five samples prepared per tooth, one remained untreated and served as a baseline control. Groups A and B were pretreated with artificial saliva and groups C and D with human saliva. Groups A and C were treated with amine fluoride and groups B and D with sodium fluoride. After treatment, samples were brushed with 25, 50, and 75 brushing strokes. The amount of KOH-soluble fluoride formed on the enamel samples was measured at baseline, after application, and after 25, 50, and 75 brushing strokes. Fluoride uptake was calculated by unpaired t-tests and fluoride retention by paired t-tests. Results. No statistically significant differences in the KOH-soluble fluoride uptake of the groups that were pretreated (A vs B and C vs D) or treated equally (A vs C and B vs D) were observed. Retention of the KOH-soluble fluoride in the brushed samples was higher when the samples were pretreated with human saliva and treated with sodium fluoride than when the samples were pretreated with artificial saliva and treated with sodium fluoride. Conclusions. The fluoride compound and the acquired human in vitro pellicle have no influence on the uptake of KOH-soluble fluoride, but show a significant influence on abrasion resistance.