Laser and fluoride treatments have been shown to inhibit enamel demineralization in the laboratory. However, the intra-oral effects of this association have not been tested. This study assessed in situ the effect of a Transversely Excited Atmospheric CO2 laser (lambda = 9.6 mum) and the use of pressure fluoridated dentifrice on enamel demineralization. During two 14-day phases, 17 volunteers wore palatal appliances containing human enamel slabs assigned to treatment groups, as follows: (1) non-fluoride dentifrice, (2) CO2 laser irradiation plus non-fluoride dentifrice, (3) fluoride dentifrice, and (4) CO2 laser irradiation plus fluoride dentifrice. A 20% sucrose solution was dripped onto the slabs 8 times per day. The specimens treated with laser and/or fluoridated dentifrice presented a significantly lower mineral loss when compared with those from the non-fluoride dentifrice group. The results suggested that CO2 laser treatment of enamel inhibits demineralization in the human mouth, being more effective when associated with fluoride.