This study evaluated the protective effect of TiF varnish, after pre-treatment with proantocyanidin or chlorhexidine, on the progression of erosive dentin loss (EDL), under the presence or absence of the demineralized organic matrix (DOM). Bovine root dentin samples were eroded for 30 min (0.1% citric acid, pH 2.5) and the loss was measured by profilometry. Half of them were subjected to the DOM removal using collagenase for 4 days, while the other half remained immersed in water. The removal of DOM was checked by profilometry. Samples were divided into 24 groups (n = 15) according to the factors: 1- With or without DOM; 2- Pre-treatment with 0.012% chlorhexidine gel, 10% proanthocyanidin gel or untreated for 1 min; 3-Final Treatment with TiF varnish, NaF varnish, placebo varnish or untreated for 6 h. The samples were submitted to a pH cycling for 5 days: 0.1% citric acid (4 × 90s/day) and artificial saliva between the challenges. The final profile was obtained for the calculation of EDL (μm, three-way ANOVA/Tukey test). When DOM was preserved, the EDL was lower compared to the condition without DOM (7.08 ± 4.03 μm and 9.80 ± 3.79 μm, p < 0.001, respectively), regardless of the treatments. The pre-treatment had no influence on the progression of EDL (p = 0.637), while the final treatment (TiF varnish only, 6.77 ± 4.08 μm) was effective in reducing the progression of EDL (NaF varnish: 9.52 ± 4.02 μm; Placebo varnish: 8.64 ± 4.06 μm and no treatment: 8.80 ± 3.95 μm). It can be concluded that DOM has important protective effect on the progression of EDL. TiF was the unique treatment capable of reducing EDL progression, regardless of the pre-treatment of DOM.Copyright © 2020 Elsevier Ltd. All rights reserved.