This study compared the progression of sealed initial enamel lesions penetrated with a fissure sealant (Helioseal, Vivadent) or various adhesives (Heliobond, Excite, Vivadent; Resulcin, Merz; Solobond M, Voco; Prompt L-Pop, 3M-ESPE) after exposure to a demineralizing solution, in vitro. From 27 bovine teeth, 54 enamel specimens were prepared and covered with nail varnish (control), thus obtaining three windows for treatment. After demineralization (pH 5.0; 14 days), two of the windows (A, B) were etched with phosphoric acid (20%; 5 seconds); whereas, the third area served as the control (C). The specimens were divided randomly into six groups (n=9), and the material was applied (90 seconds) either once (A) or twice (B). Light-curing followed each application. Half of the area of each specimen window was then covered with nail varnish, and the samples were again stored in the demineralizing solution (pH 5.0; 14 days). The specimens were cut perpendicular to the surface, and both enamel slabs were studied after infiltration using a fluorescent, low viscous resin (VIRIN) and confocal microscopy (CLSM). Lesion depths were calculated (ImageJ) from the surface to that point in the lesion where the grey values clearly changed to a darker grey. After demineralization, mean lesion depths (SD) (14 days) were measured at 105 (21) microm. The second demineralization led to a mean progression of the lesion depths of 52 (31)%. Adper Prompt L-Pop and Solobond M could not significantly prevent lesion progression after a single application (p > 0.05; t-test); however, the second application of Solobond M significantly decreased lesion progression (p < 0.05; t-test). Helioseal, Heliobond, Resulcin Monobond and Excite showed significantly better inhibition of the demineralization compared to the other materials (p < 0.05; Bonferroni). It can be concluded that the penetration of adhesives into initial lesions inhibited a further demineralization in vitro.